WO2010119343A2 - Compositions for immunising against staphylococcus aureus - Google Patents

Compositions for immunising against staphylococcus aureus Download PDF

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Publication number
WO2010119343A2
WO2010119343A2 PCT/IB2010/000998 IB2010000998W WO2010119343A2 WO 2010119343 A2 WO2010119343 A2 WO 2010119343A2 IB 2010000998 W IB2010000998 W IB 2010000998W WO 2010119343 A2 WO2010119343 A2 WO 2010119343A2
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antigen
seq
amino acids
acid sequence
amino acid
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PCT/IB2010/000998
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French (fr)
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WO2010119343A8 (en
WO2010119343A3 (en
Inventor
Fabio Bagnoli
Massimilliano Biagini
Luigi Fiaschi
Guido Grandi
Ravi Mishra
Nathalie Norais
Maria Scarselli
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Novartis Ag
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First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=42734088&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=WO2010119343(A2) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority to CA2758490A priority Critical patent/CA2758490C/en
Priority to MX2011010735A priority patent/MX2011010735A/en
Priority to MX2015006258A priority patent/MX363222B/en
Priority to SG2011072832A priority patent/SG175092A1/en
Priority to EP10723299A priority patent/EP2419129A2/en
Priority to RU2011146016/15A priority patent/RU2508126C2/en
Priority to JP2012505252A priority patent/JP5830009B2/en
Priority to CN201810736674.6A priority patent/CN109248313B/en
Priority to US13/264,369 priority patent/US8679505B2/en
Priority to AU2010238255A priority patent/AU2010238255B2/en
Priority to CN2010800274105A priority patent/CN102647999A/en
Priority to NZ595689A priority patent/NZ595689A/en
Priority to BRPI1013780A priority patent/BRPI1013780B8/en
Application filed by Novartis Ag filed Critical Novartis Ag
Publication of WO2010119343A2 publication Critical patent/WO2010119343A2/en
Publication of WO2010119343A3 publication Critical patent/WO2010119343A3/en
Priority to US13/234,077 priority patent/US8632783B2/en
Priority to IL215593A priority patent/IL215593A0/en
Publication of WO2010119343A8 publication Critical patent/WO2010119343A8/en
Priority to US14/171,537 priority patent/US9205142B2/en
Priority to PH12015500092A priority patent/PH12015500092A1/en
Priority to US14/961,772 priority patent/US9642904B2/en
Priority to US15/589,834 priority patent/US10195263B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/085—Staphylococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/305—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F)
    • C07K14/31—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • This invention relates to antigens derived from S.aureus and to their use in immunisation.
  • Staphylococcus aureus is a Gram-positive spherical bacterium. Annual US mortality exceeds that of any other infectious disease, including HIV/ AIDS, and S.aureus is the leading cause of bloodstream, lower respiratory tract, skin & soft tissue infections. There is currently no authorised vaccine.
  • Reference 1 reports that the "V710" vaccine from Merck and Intercell is undergoing a phase 2/3 trial on patients undergoing cardiothoracic surgery.
  • the V710 vaccine is based on a single antigen, IsdB [2], a conserved iron-sequestering cell-surface protein.
  • S.aureus causes a range of illnesses from minor skin infections to life-threatening diseases such as pneumonia, meningitis, osteomyelitis, bacteremia, endocarditis, toxic shock syndrome, organ abscesses and septicemia.
  • the bacterium has multiple virulence factors which are differentially expressed during different phases of its life cycle, and so a vaccine which can prevent one disease might not prevent another.
  • the V710 vaccine may be effective against hematic spread of the S.aureus, but may be ineffective against pneumonia and may not elicit any opsonic activity.
  • One aim of the invention is to provide vaccines which can protect against hematic spread and pneumonia, and which may also elicit an opsonic response.
  • the inventors have identified various S.aureus polypeptides that are useful for immunisation, either alone or in combination. These polypeptides may be combined with S.aureus saccharides or other S.aureus polypeptides.
  • the antigens are useful in S.aureus vaccines but may also be used as components in vaccines for immunising against multiple pathogens.
  • the inventors have identified the following 36 polypeptides: clfA, clfB, coA, eap, ebhA, ebpS, efb, emp, esaC, esxA, esxB, FnBA, FnBB, HIa, hlgB, hlgC, isdA, isdB, isdC, isdG, isdH, isdl, lukD, lukE, lukF, lukS, nuc, sasA, sasB, sasC, sasD, sasF, sdrC, sdrD, spa, and sdrE2.
  • the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a clfA antigen; (2) a clfB antigen; (3) a coA antigen; (4) a eap antigen; (5) a ebhA antigen; (6) a ebpS antigen; (7) a efb antigen; (8) a emp antigen; (9) a esaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) a FnBA antigen; (13) a FnBB antigen; (14) a HIa antigen; (15) a hlgB antigen; (16) a hlgC antigen; (17) a isdA antigen;
  • antigens are preferably selected from a subset of 16 of the 36 polypeptides, namely: clfA, clfB, emp, esaC, esxA, esxB, hla, isdA, isdB, isdC, sasD, sasF, sdrC, sdrD, spa, and sdrE2.
  • an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of these sixteen antigens.
  • the inventors have also identified the following 128 polypeptides: staOOl, sta002, staOO3, staOO4, staOO5, sta006, sta007, staOO8, sta009, staOlO, staOl l, staO12, staO13, staO14, staO15, staOl ⁇ , staO17, staO18, staO19, sta020, staO21, staO22, staO23, staO24, staO25, staO26, staO27, staO28, staO29, staO3O, staO31, staO32, staO33, staO34, staO35, staO36, staO37, staO38, staO39, staO4O, staO41, staO42, staO43, staO44, staO45, staO46, staO47, staO48, staO49, staO5O, staO51, staO52, staO53, staO54, staO55, staO56, staO57, staO58, staO59, sta0
  • the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a staOOl antigen; (2) a sta002 antigen; (3) a staOO3 antigen; (4) a sta004 antigen; (5) a staOO5 antigen; (6) a sta006 antigen; (7) a sta007 antigen; (8) a staOO8 antigen; (9) a sta009 antigen; (10) a staOlO antigen; (11) a staOl l antigen; (12) a staO12 antigen; (13) a staO13 antigen; (14) a staO14 antigen; (15) a staO15 antigen; (16) a staOl ⁇ antigen; (17) a staO17 antigen; (18) a staO18 antigen; (19)
  • a staO56 antigen a staO56 antigen; (57) a staO57 antigen; (58) a staO58 antigen; (59) a staO59 antigen; (60) a sta060 antigen; (61) a staO ⁇ l antigen; (62) a staO62 antigen; (63) a staO63 antigen; (64) a staO64 antigen; (65) a staO65 antigen; (66) a staO66 antigen; (67) a staO67 antigen; (68) a staO68 antigen; (69) a staO69 antigen; (70) a sta070 antigen; (71) a staO71 antigen; (72) a staO72 antigen; (73) a staO73 antigen; (74) a staO74 antigen; (75) a staO75 antigen; (76) a staO76 antigen; (77) a staO77 antigen; (78) a staO78 antigen; (79) a
  • a preferred subset of 113 antigens omits (81) and (107) to (120) from this list.
  • a subset of 27 of the 128 polypeptides is referred to herein as 'the third antigen group', namely: staOOl, sta002, sta003, sta004, staOO5, sta006, sta007, staOO8, sta009, staOlO, staO19, staO28, sta040, staO49, staO57, staO64, staO73, staO95, staO98, stalOl, stalO5, NW l, NW 6, NWJ, NWJ, NW 9 and NW lO.
  • the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more ⁇ i.e. 2, 3, 4, 5, 6 or more) antigens selected from the third antigen group.
  • the 101 antigens that are in the second antigen group but not in the third antigen group are referred to herein as 'the fourth antigen group'.
  • a preferred subset of 86 antigens omits (81) and (107) to (120) from the above list.
  • the second antigen group thus consists of a combination of the third and fourth antigen groups.
  • a subset of 8 of the 128 polypeptides is referred to herein as 'the fifth antigen group', namely: staOO4, sta006, sta007, staOl l, staO28, sta060, staO98 and stal l2.
  • the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more ⁇ i.e. 2, 3, 4, 5, 6 or more) antigens selected from the fifth antigen group.
  • a combination of antigens said combination comprising two or more ⁇ i.e. 2, 3, 4, 5, 6 or more) antigens selected from the fifth antigen group.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 630 pairs.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 8128 pairs.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 6328 pairs.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 351 pairs.
  • Within the 101 antigens of the fourth antigen group there are 5050 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 5050 pairs.
  • Within the preferred 86 antigens of the fourth antigen group there are 3655 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention.
  • the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 3655 pairs.
  • a composition in one embodiment, includes at least one antigen (i.e. 1, 2, 3, 4, 5, 6 or more) selected from the first antigen group and at least one antigen (i.e. 1, 2, 3, 4, 5, 6 or more) selected from the second antigen group.
  • Antigens from the first antigen group may be selected from the preferred subset of 16 antigens, and antigens from the second antigen group may be selected from the third antigen group or the fifth antigen group.
  • the invention also provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a clfA antigen; (2) a clfB antigen; (3) a sdrE2 antigen; (4) a sdrC antigen; (5) a
  • This group of 18 antigens is sometimes referred to herein as the 'sixth antigen group'.
  • the invention also provides an immunogenic composition
  • a combination of antigens comprising two or more (i.e. 2, 3, 4 or 5) antigens selected from the group consisting of: (1) a esxA antigen; (2) a esxB antigen; (3) a staOO ⁇ antigen; (4) a hla antigen; and/or (5) a staOl 1 antigen.
  • the composition may also include an adjuvant e.g. an aluminium hydroxide adjuvant.
  • Advantageous combinations of the invention are those in which two or more antigens act synergistically. Thus the protection against S. aureus disease achieved by their combined administration exceeds that expected by mere addition of their individual protective efficacy.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen and a isdC antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an SdrDE hybrid with an sdrE2 antigen downstream of a sdrD antigen.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen and a sdrC antigen.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a clfA antigen.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a staOl 1 antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a staOO ⁇ antigen.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a hla antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
  • the Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a esxA antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen and a hla antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a esxA antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen and a staOl l antigen.
  • the esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen and a staOl l antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a spa antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a isdA antigen, a staOO ⁇ antigen, a staOl 1 antigen and a spa antigen.
  • the esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • the spa antigen may be a fragment of a full- length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a HIa antigen, a staOO ⁇ antigen and a staOl l antigen.
  • the esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a sdrE2 antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen and a hla antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a hla antigen, a isdA antigen, a staOO ⁇ antigen and a staOl 1 antigen.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • the Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen and a isdA antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a hla antigen.
  • the Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a HIa antigen, a staOO ⁇ antigen and a staOl l antigen.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a esxA antigen and a esxB antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid with an esxB antigen downstream of an esxA antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen and a staOO ⁇ antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • An immunogenic composition comprising a spa antigen, a staOO ⁇ antigen and a staOl l antigen.
  • the spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a isdA antigen, a staOO ⁇ antigen and a staOl l antigen.
  • the esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • An immunogenic composition comprising a staOO ⁇ antigen and a staOl 1 antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen, a isdA antigen and a clfB antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • the clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163.
  • An immunogenic composition comprising a staOO ⁇ antigen, a staOl l antigen and a staO19 antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen, a hla antigen and a clfB antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a staOO ⁇ antigen, a staOl l antigen, a staO19 antigen, and a hla antigen.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen, a staOl 1 antigen and a clfB antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163.
  • An immunogenic composition comprising a spa antigen, a esxA antigen, a esxB antigen, a staOO ⁇ antigen and a staOl 1 antigen.
  • the spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
  • the esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
  • An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a esxB antigen.
  • the sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen, a staOl l antigen and a staO19 antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOO ⁇ antigen, a isdA antigen and a sdrD antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • the sdrD antigen may be a fragment of a full-length sdrD antigen e.g. SEQ ID NO: 156.
  • An immunogenic composition comprising a esxA antigen, a esxB antigen, and a isdA antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
  • the isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
  • An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen, a esxA antigen and a esxB antigen.
  • the esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid with an esxB antigen downstream of an esxA antigen.
  • An immunogenic composition comprising a HIa antigen, a spa antigen, a staOO ⁇ antigen and a staOl 1 antigen.
  • the HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
  • the spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
  • any of these 37 compositions may include additional staphylococcal antigens, and these further antigens can be polypeptides and/or saccharides.
  • these further antigens can be polypeptides and/or saccharides.
  • they can usefully also include one or more S. ⁇ ureus capsular saccharide co ⁇ jugate(s) e.g. against a serotype 5 and/or a serotype 8 strain.
  • S. ⁇ ureus capsular saccharide co ⁇ jugate(s) e.g. against a serotype 5 and/or a serotype 8 strain.
  • the inclusion of one or both such conjugates is particularly useful for combinations (8), (10), (20), (23), (25), (31) and (37).
  • these 37 compositions include no additional staphylococcal polypeptide antigens, hi other embodiments, these 37 compositions include no additional staphylococcal antigens, hi other embodiments, these 37 compositions include no additional antigens.
  • the invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 151; and/or (b) comprising a fragment of at least 'n' consecutive amino acids from amino acids 1-97 of SEQ ID NO: 151 and at least 'n' consecutive amino acids from amino acids 104- 207 of SEQ ID NO: 151, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • the invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 152; and/or (b) comprising a fragment of at least 'n' consecutive amino acids from amino acids 1-104 of SEQ ID NO: 152 and at least 'n' consecutive amino acids from amino acids 111-207 of SEQ ID NO: 152, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • polypeptides can elicit antibodies (e.g. when administered to a human) which recognise both the wild-type staphylococcal protein comprising SEQ ID NO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO: 11.
  • the immune response will recognise both of antigens esxA and esxB.
  • Preferred fragments of (b) provide an epitope from SEQ ID NO: 10 and an epitope from SEQ ID NO: 11.
  • the invention also provides an immunogenic composition comprising a combination of such a protein and an adjuvant, such as an aluminium hydroxide adjuvant.
  • the invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 241; and/or (b) comprising both a fragment of at least 'n' consecutive amino acids from amino acids 1-96 of SEQ ID NO: 241 and a fragment of at least 'n' consecutive amino acids from amino acids 103-205 of SEQ ID NO: 241, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or
  • polypeptides e.g. SEQ ID NO: 250
  • antibodies e.g. when administered to a human
  • the immune response will recognise both of antigens esxA and esxB.
  • Preferred fragments of (b) provide an epitope from SEQ ID NO: 10 and an epitope from SEQ ID NO: 11.
  • the invention also provides an immunogenic composition comprising a combination of such a protein and an adjuvant, such as an aluminium hydroxide adjuvant.
  • the invention also provides a polypeptide comprising a staphylococcal hemolysin sequence, wherein the sequence does not include a sequence having at least 90% identity to SEQ ID NO: 217 but can elicit antibodies which can kill staphylococci.
  • the polypeptide may have a first sequence having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 218 and a second sequence having 80% or more identity (e.g.
  • SEQ ID NO: 219 wherein the first and second sequences are either directly joined or are joined by an intervening amino acid sequence having fewer than 40 amino acids (e.g. ⁇ 35 amino acids, ⁇ 30 amino acids, ⁇ 25 amino acids, ⁇ 20 amino acids, ⁇ 15 amino acids, ⁇ 10 amino acids, ⁇ 5 amino acids).
  • SEQ ID NOs: 189 and 216 are examples of such polypeptides, in which the first and second sequences are joined by a tetrapeptide PSGS sequence (SEQ ID NO: 225).
  • the invention also provides an immunogenic composition comprising a StaOl l antigen and a Ca + * ion.
  • the antigen and Ca ⁇ ion may form a complex e.g. atoms in the antigen may coordinate the Ca +* ion.
  • the immunogenic composition may also include an adjuvant.
  • the invention also provides a oligomer of a StaOl l antigen, and also immunogenic compositions comprising such oligomers.
  • the oligomer can be a dimer, trimer, tetramer, pentamer, hexamer, heptamer, octamer or higher.
  • An oligomer may comprise a Ca +4 ⁇ ion, and a composition comprising StaOl 1 oligomers may comprise 5-50OmM Ca + * ions.
  • immunogenic compositions may include one or more of the following S. aureus antigens (or antigens comprising immunogenic fragment(s) thereof) to enhance the efficacy against S.aureus of an immune response elicited by the composition [e.g. see references 3-10]:
  • the individual antigens identified in the antigen groups of the invention may be used in combination with conjugated saccharide antigens.
  • an immunogenic composition comprising a combination of:
  • antigen(s) selected from the first, second, third or fourth antigen groups (as defined above); and (2) one or more conjugates of a S.aureus exopolysaccharide and a carrier protein.
  • a conjugate used in component (2) of this combination includes a saccharide moiety and a carrier moiety.
  • the saccharide moiety is from the exopolysaccharide of S.aureus, which is a poly-N- acetylglucosamine (PNAG).
  • PNAG poly-N- acetylglucosamine
  • the saccharide may be a polysaccharide having the size that arises during purification of the exopolysaccharide from bacteria, or it may be an oligosaccharide achieved by fragmentation of such a polysaccharide e.g. size can vary from over 40OkDa to between 75 and 40OkDa, or between 10 and 75kDa, or up to 30 repeat units.
  • the saccharide moiety can have various degrees of N-acetylation and, as described in reference 11, the PNAG may be less than 40% N-acetylated (e.g. less than 35, 30, 20, 15, 10 or 5% N-acetylated; deacetylated PNAG is also known as dPNAG).
  • Deacetylated epitopes of PNAG can elicit antibodies that are capable of mediating opsonic killing.
  • the PNAG may or may not be O-succinylated e.g. it may be O-succinylated on fewer less than 25, 20, 15, 10, 5, 2, 1 or 0.1% of residues.
  • the invention also provides an immunogenic composition comprising a combination of:
  • antigen(s) selected from the first, second, third or fourth antigen groups.
  • a conjugate used in component (2) of this combination includes a saccharide moiety and a carrier moiety.
  • the saccharide moiety is from the capsular saccharide of a S.aureus.
  • the saccharide may be a polysaccharide having the size that arises during purification of capsular polysaccharide from bacteria, or it may be an oligosaccharide achieved by fragmentation of such a polysaccharide.
  • Capsular saccharides may be obtained from any suitable strain of S.aureus (or any bacterium having a similar or identical saccharide), such as from a type 5 and/or a type 8 S.aureus strain and/or a type
  • S.aureus strain Most strains of infectious S.aureus contain either Type 5 or Type 8 capsular saccharides. Both have FucNAcp in their repeat unit as well as ManNAcA which can be used to introduce a sulfhydryl group for linkage.
  • the repeating unit of the Type 5 saccharide is ⁇ 4)- ⁇ -D- Man NAcA-(I ⁇ 4)- ⁇ -L-FucNAc(3OAc)-(l — » ⁇ )- ⁇ -D-FucNAc-(l ⁇
  • the repeating unit of the Type 8 saccharide is ⁇ 3)- ⁇ -D-ManNAcA(4OAc)-(l ⁇ 3)- ⁇ -L-FucNAc(l ⁇ 3)- ⁇ -D-FucNAc(l ⁇ .
  • the type 336 saccharide is a ⁇ -linked hexosamine with no O-acetylation [12,13] and is cross-reactive with antibodies raised against the 336 strain (ATCC 55804). A combination of a type 5 and a type 8 saccharide is typical, and a type 336 saccharide may be added to this pairing [14].
  • the invention also provides an immunogenic composition comprising a combination of:
  • antigen(s) selected from the first, second, third or fourth antigen groups;
  • the carrier moiety in these conjugates will usually be a protein, but usually not one of the antigens of (1).
  • Typical carrier proteins are bacterial toxins, such as diphtheria or tetanus toxins, or toxoids or mutants or fragments thereof.
  • the CRM 197 diphtheria toxin mutant [15] is useful.
  • Other suitable carrier proteins include the N.
  • the carrier protein is a S.aureus protein, such as an antigen selected from the first, second, third or fourth antigen groups.
  • each conjugate may use the same carrier protein or a different carrier protein.
  • Conjugates may have excess carrier (w/w) or excess saccharide (w/w).
  • a conjugate may include substantially equal weights of each.
  • the carrier molecule may be covalently conjugated to the carrier directly or via a linker.
  • Direct linkages to the protein may be achieved by, for instance, reductive animation between the saccharide and the carrier, as described in, for example, references 35 and 36.
  • the saccharide may first need to be activated e.g. by oxidation.
  • Linkages via a linker group may be made using any known procedure, for example, the procedures described in references 37 and 38.
  • a preferred type of linkage is an adipic acid linker, which may be formed by coupling a free -NH 2 group ⁇ e.g.
  • linkage is a carbonyl linker, which may be formed by reaction of a free hydroxyl group of a saccharide CDI [41, 42] followed by reaction with a protein to form a carbamate linkage.
  • linkers include ⁇ - propionamido [43], nitrophenyl-ethylamine [44], haloacyl halides [45], glycosidic linkages [46], 6- aminocaproic acid [47], ADH [48], C 4 to Ci 2 moieties [49], etc.
  • Carbodiimide condensation can also be used [50].
  • PNAG conjugates may be prepared in various ways e.g. by a process comprising: a) activating the PNAG by adding a linker comprising a maleimide group to form an activated PNAG; b) activating the carrier protein by adding a linker comprising a sulphydryl group to form an activated carrier protein; and c) reacting the activated PNAG and the activated carrier protein to form a PNAG-carrier protein conjugate; or by a process comprising a) activating the PNAG by adding a linker comprising a sulphydryl group to form an activated PNAG; b) activating the carrier protein by adding a linker comprising a maleimide group to form an activated carrier protein; and c) reacting the activated PNAG and the activated carrier protein to form a PNAG-carrier protein conjugate; or by a process comprising a) activating the PNAG by adding a linker comprising a sulphy
  • the individual antigens identified in the antigen groups of the invention may be used as carrier proteins for exopolysaccharides, to form a covalent conjugate.
  • the invention provides an immunogenic composition comprising a conjugate of (1) an antigen selected from the first, second, third and fourth antigen groups and (2) a S. aureus exopolysaccharide.
  • the invention also provides an immunogenic composition comprising a conjugate of (1) an antigen selected from the first, second, third and fourth antigen groups and (2) a S.aureus capsular saccharide. Further characteristics of such conjugates are described above. These conjugates may be combined with any of the antigens disclosed herein.
  • the individual antigens identified in the antigen groups of the invention may be used in combination with non-staphylococcal antigens, and in particular with antigens from bacteria associated with nosocomial infections.
  • an immunogenic composition comprising a combination of:
  • antigen(s) selected from the first, second, third and fourth antigen groups (as defined above);
  • antigen(s) selected from the group consisting of: Clostridium difficile; Pseudomonas aeruginosa; Candida albicans; and extraintestinal pathogenic Escherichia coli.
  • the 'clfA' antigen is annotated as 'clumping factor A'.
  • clfA is SAOUHSC 00812 and has amino acid sequence SEQ ID NO: 1 (GI: 88194572).
  • nwmn_0756 GI: 151220968.
  • Useful clfA antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 1 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 1; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 1, wherein 'n' is 7 or more ⁇ e.g.
  • These clfA proteins include variants of SEQ ID NO: 1.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 1.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 1 while retaining at least one epitope of SEQ ID NO: 1.
  • the final 368 C-terminal amino acids of SEQ ID NO: 1 can usefully be omitted.
  • the first 39 N-terminal amino acids of SEQ ID NO: 1 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • SEQ ID NO: 224 is a useful fragment of SEQ ID NO: 1 ('CIfA 40-559 ')- This fragments omits the long repetitive region towards the C-terminal of SEQ ID NO: 1.
  • the 'clfB' antigen is annotated as 'clumping factor B'.
  • clfB is SAOUHSC 02963 and has amino acid sequence SEQ ID NO: 2 (GI:88196585). In the Newman strain it is nwmn_2529 (GI: 151222741).
  • Useful clfB antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 2 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%,
  • SEQ ID NO: 2 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 2; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
  • SEQ ID NO: 2 wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
  • These clfB proteins include variants of SEQ ID NO: 2.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 2.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ
  • SEQ ID NO: 2 while retaining at least one epitope of SEQ ID NO: 2.
  • the final 40 C-terminal amino acids of SEQ ID NO: 2 can usefully be omitted.
  • the first 44 N-terminal amino acids of SEQ ID NO: 2 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • ClfB is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc.
  • SEQ ID NO: 163 is a useful fragment of SEQ ID NO: 2 ('CIfB 45-552 '). This fragment includes the most exposed domain of CIfB and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins.
  • CIfB 45-360 also known as CLf ⁇ -N12; SEQ ID NO: 196
  • CIfB 2I2-542 also known as CLfB-N23; SEQ ID NO: 197
  • CIfB 360-542 also known as CLfB-N3; SEQ ID NO: 198.
  • the 'coA' antigen is annotated as 'coagulase Coa'.
  • coA is SAOUHSC 00192 and has amino acid sequence SEQ ID NO: 3 (GI:88194002).
  • SEQ ID NO: 3 GI:88194002
  • nwmn_0166 GI: 151220378.
  • Useful coA antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 3 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 3 SEQ ID NO: 3; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 3, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • coA proteins include variants of SEQ ID NO: 3.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 3.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'eap' antigen is annotated as 'MHC class II analog protein', hi the NCTC 8325 strain eap is SAOUHSC 02161 and has amino acid sequence SEQ ID NO: 4 (GI: 88195840). hi the Newman strain it is nwmn_1872 (GI: 151222084).
  • Useful eap antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 4 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 4 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 4, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • eap proteins include variants of SEQ ID NO: 4.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 4.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'ebhA' antigen is annotated as 'EbhA'.
  • ebhA is SAOUHSC O 1447 and has amino acid sequence SEQ ID NO: 5 (GI: 88195168).
  • Useful ebhA antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 5 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 5; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 5, wherein 'n' is 7 or more (e.g.
  • ebhA proteins include variants of SEQ ID NO: 5.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 5.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 5 while retaining at least one epitope of SEQ ID NO: 5.
  • the first 39 N- terminal amino acids of SEQ ID NO: 5 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'ebpS' antigen is annotated as 'elastin binding protein EbpS'.
  • ebpS is SAOUHSC 01501 and has amino acid sequence SEQ ID NO: 6 (GI: 88195217).
  • SEQ ID NO: 6 GI: 88195217).
  • Newman strain it is nwmn_1389 (GI:151221601).
  • Useful ebpS antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 6 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 6 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 6, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • ebpS proteins include variants of SEQ ID NO: 6.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 6.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • SEQ ID NO: 165 is a useful fragment of SEQ ID NO: 6 ( ⁇ bpSi.i 98 '). This fragment includes the most exposed domain of EbpS and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. efb
  • the 'efb' antigen is annotated as 'f ⁇ brinogen-binding protein truncated'.
  • efb is SAOUHSC Ol 114 and has amino acid sequence SEQ ID NO: 7 (GI: 88194860).
  • SEQ ID NO: 7 In the Newman strain it is nwmn l 069 (GI: 151221281 ).
  • Useful efb antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 7 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 7 comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 7, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • efb proteins include variants of SEQ ID NO: 7.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 7.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • the 'emp' antigen is annotated as 'extracellular matrix and plasma binding protein'.
  • emp is SAOUHSC 00816 and has amino acid sequence SEQ ID NO: 8 (GI: 88194575).
  • nwmn_0758 GI: 151220970.
  • Useful emp antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 8 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 8 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 8, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more.
  • These emp proteins include variants of SEQ ID NO: 8.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 8.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • SEQ ID Nos: 190, 191, 192 and 193 are useful fragments of SEQ ID NO: 8 ( ⁇ mp 35-34 o ⁇ ⁇ mp 27-334 ⁇ 'Emp 35-334 ' and 'Emp 27-147 ', respectively).
  • the 'esaC antigen is annotated as 'esaC.
  • esaC is SAOUHSC 00264 and has amino acid sequence SEQ ID NO: 9 (GI:88194069).
  • Useful esaC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 9 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 9 SEQ ID NO: 9; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 9, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • esaC proteins include variants of SEQ ID NO: 9.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 9.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'esxA' antigen is annotated as 'protein'.
  • esxA is SAOUHSC 00257 and has amino acid sequence SEQ ID NO: 10 (GI:88194063).
  • Useful esxA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 10 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 10 SEQ ID NO: 10
  • esxA proteins include variants of SEQ ID NO: 10.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 10.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'esxB' antigen is annotated as 'esxB'.
  • esxB is SAOUHSC 00265 and has amino acid sequence SEQ ID NO: 11 (GI:88194070).
  • Useful esxB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 11 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%,
  • SEQ ID NO: 11 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 11; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
  • SEQ ID NO: 11 wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
  • esxB proteins include variants of SEQ ID NO: 11.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 11.
  • Other preferred fragments lack one or more amino acids
  • the 1 FnBA' antigen is annotated as 'fibronectin-binding protein A precursor FnBPA'.
  • FnBA is SAOUHSC 02803 and has amino acid sequence SEQ ID NO: 12 (GL88196438).
  • nwmn_2399 GI: 151222611. Proteomic analysis has revealed that this protein is secreted or surface-exposed.
  • Useful FnBA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 12 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 12; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 12, wherein 'n' is 7 or more (e.g.
  • FnBA proteins include variants of SEQ ID NO: 12.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 12.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 12 while retaining at least one epitope of SEQ ID NO: 12.
  • the final 37 C- terminal amino acids of SEQ ID NO: 12 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • FnBA is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc.
  • SEQ ID NOs: 166 ('FnBAj -511 ') and 167 ('FnBA 5 I 2-953 ') are useful fragments of SEQ ID NO: 12. These fragments are more easily used at an industrial scale.
  • the 'FnBB' antigen is annotated as 'fibronectin binding protein B FnBPB'.
  • FnBB is SAOUHSC 02802 and has amino acid sequence SEQ ID NO: 13 (GI:88196437).
  • nwmn_2397 GI: 151222609).
  • Useful FnBB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 13 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 13; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 13, wherein 'n' is 7 or more (e.g.
  • FnBB proteins include variants of SEQ ID NO: 13.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 13.
  • Other preferred fragments lactone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 13 while retaining at least one epitope of SEQ ID NO: 13.
  • the final 37 C- terminal amino acids of SEQ ID NO: 13 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'HIa' antigen is the 'alpha-hemolysin precursor' also known as 'alpha toxin' or simply 'hemolysin'.
  • HIa is SAOUHSC Ol 121 and has amino acid sequence SEQ ID NO: 14 (GI: 88194865). In the Newman strain it is nwmn_1073 (GI:151221285).
  • HIa is an important virulence determinant produced by most strains of S.aureus, having pore-forming and haemolytic activity. Anti-Hla antibodies can neutralise the detrimental effects of the toxin in animal models, and HIa is particularly useful for protecting against pneumonia.
  • Useful HIa antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 14 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 14; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 14, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • HIa proteins include variants of SEQ ID NO: 14.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 14.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 14 while retaining at least one epitope of SEQ ID NO: 14.
  • the first 26 N-terminal amino acids of SEQ ID NO: 14 can usefully be omitted (e.g. to give SEQ ID NO: 231).
  • Truncation at the C- terminus can also be used e.g. leaving only 50 amino acids (residues 27-76 of SEQ ID NO: 14) [52].
  • Other fragments omit one or more protein domains.
  • HIa' s toxicity can be avoided in compositions of the invention by chemical inactivation (e.g. using formaldehyde, glutaraldehyde or other cross-linking reagents). Instead, however, it is preferred to use mutant forms of HIa which remove its toxic activity while retaining its immunogenicity. Such detoxified mutants are already known in the art.
  • residue 61 may not be histidine, and may instead be e.g. He, VaI or preferably Leu.
  • SEQ ID NO: 150 is the mature mutant Hla-H35L sequence (i.e. SEQ ID NO: 231 with a H35L mutation) and a useful HIa antigen comprises SEQ ID NO: 150.
  • Another useful mutation replaces a long loop with a short sequence e.g. to replace the 39mer at residues 136-174 of SEQ ID NO: 14 with a tetramer such as PSGS (SEQ ID NO: 225), as in SEQ ID NO: 189 (which also includes the H35L mutation) and SEQ ID NO: 216 (which does not include the H35L mutation).
  • Another useful mutation replaces residue YlOl e.g.
  • Another useful mutation replaces residue D 152 e.g. with a leucine (SEQ ID NO: 243).
  • Another useful mutant replaces residues H35 and YlOl e.g. with a leucine (SEQ ID NO: 244).
  • Another useful mutant replaces residues H35 and D152 e.g. with a leucine (SEQ ID NO: 245).
  • SEQ ID NOs: 160, 161 & 194 are three useful fragments of SEQ ID NO: 14 ('HIa 27-76 ', 'HIa 27-89 ' and 'HIa 27-79 ', respectively).
  • SEQ ID NOs: 158, 159 and 195 are the corresponding fragments from SEQ ID NO: 150.
  • SEQ ID NO: 232 One useful HIa sequence is SEQ ID NO: 232, which was used in the examples. It has a N-terminal Met, then an Ala-Ser dipeptide from the expression vector, then SEQ ID NO: 150 (from NCTC8325 strain). It is encoded by SEQ ID NO: 233.
  • MgB One useful HIa sequence is SEQ ID NO: 232, which was used in the examples. It has a N-terminal Met, then an Ala-Ser dipeptide from the expression vector, then SEQ ID NO: 150 (from NCTC8325 strain). It is encoded by SEQ ID NO: 233.
  • the 'hlgB' antigen is annotated as 'leukocidin f subunit precursor HIgB'.
  • hlgB is SAOUHSC 02710 and has amino acid sequence SEQ ID NO: 15 (GI:88196350).
  • Useful hlgB antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 15 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 15; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 15, wherein 'n' is 7 or more (e.g.
  • hlgB proteins include variants of SEQ ID NO: 15.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 15.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 15 while retaining at least one epitope of SEQ ID NO: 15.
  • the first 26 N-terminal amino acids of SEQ ID NO: 15 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'hlgC antigen is annotated as 'leukocidin s subunit precursor HIgC.
  • hlgC is SAOUHSC_02709 and has amino acid sequence SEQ ID NO: 16 (GI: 88196349).
  • Useful hlgC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 16 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 16; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 16, wherein V is 7 or more (e.g.
  • hlgC proteins include variants of SEQ ID NO: 16.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 16.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 16 while retaining at least one epitope of SEQ ID NO: 16.
  • the first 29 N-terminal amino acids of SEQ ID NO: 16 can usefully be omitted.
  • Other fragments omit one or more protein domains. isdA
  • the 'isdA' antigen is annotated as 'IsdA protein'.
  • isdA is SAOUHSC O 1081 and has amino acid sequence SEQ ID NO: 17 (GI: 88194829).
  • nwmn_1041 GI: 151221253
  • Useful isdA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 17 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 17; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 17, wherein 'n' is 7 or more (e.g.
  • isdA proteins include variants of SEQ ID NO: 17.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 17.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 17 while retaining at least one epitope of SEQ ID NO: 17.
  • the final 38 C-terminal amino acids of SEQ ID NO: 17 can usefully be omitted.
  • the first 46 N-terminal amino acids of SEQ ID NO: 17 can usefully be omitted. Truncation to exclude the C-terminal 38mer of SEQ ID NO: 17 (beginning with the LPKTG motif) is also useful. Other fragments omit one or more protein domains.
  • SEQ ID NO: 157 is a useful fragment of SEQ ID NO: 17 (amino acids 40-184 of SEQ ID NO: 17; 'ISdA 4O-I84 ') which includes the natural protein's heme binding site and includes the antigen's most exposed domain. It also reduces the antigen's similarity with human proteins.
  • Other useful fragments are disclosed in references 55 and 56.
  • IsdA does not adsorb well to aluminium hydroxide adjuvants, so IsdA present in a composition may me unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate.
  • Anti-IsdA antibodies protect mice against S. aureus abscess formation and lethal challenge [57]. isdB
  • the 'isdB' antigen is annotated as 'neurofilament protein isdB'.
  • isdB is SAOUHSC 01079 and has amino acid sequence SEQ ID NO: 18 (GL88194828). IsdB has been proposed for use as a vaccine antigen on its own [2], but this may not prevent pneumonia.
  • Useful isdB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
  • SEQ ID NO: 18 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 18, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • isdB proteins include variants of SEQ ID NO: 18.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 18.
  • Other preferred fragments lack one or more amino acids (e.g.
  • compositions of the invention do not include an isdB antigen. isdC
  • the 'isdC antigen is annotated as 'protein'.
  • isdC is SAOUHSC O 1082 and has amino acid sequence SEQ ID NO: 19 (GI: 88194830).
  • Useful isdC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 19 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 19; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 19, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • isdC proteins include variants of SEQ ID NO: 19.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 19.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 19 while retaining at least one epitope of SEQ ID NO: 19.
  • the final 39 C-terminal amino acids of SEQ ID NO: 19 can usefully be omitted.
  • the first 28 N-terminal amino acids of SEQ ID NO: 19 can usefully be omitted.
  • Other fragments omit one or more protein domains. Useful fragments of IsdB are disclosed in reference 56.
  • Reference 59 discloses antigens which usefully include epitopes from both IsdB and IsdH. isdG
  • the 'isdG' antigen is annotated as 'heme-degrading monooxygenase IsdG'.
  • isdG is SAOUHSC 01089 and has amino acid sequence SEQ ID NO: 20 (GI: 88194836).
  • Useful isdG antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
  • ID NO: 20 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 20; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 20, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • isdG proteins include variants of SEQ ID NO: 20.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 20.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 20 while retaining at least one epitope of SEQ ID NO: 20.
  • Other fragments omit one or more protein domains.
  • isdH The 'isdH' antigen is annotated as 'isdH'.
  • isdH is SAOUHSC Ol 843 and has amino acid sequence SEQ ID NO: 21 (GI:88195542).
  • nwmn_1624 GI: 151221836). It has also been known as HarA.
  • Useful isdH antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 21 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 21; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 21, wherein 'n' is 7 or more (e.g.
  • isdH proteins include variants of SEQ ID NO: 21.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 21.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 21 while retaining at least one epitope of SEQ ID NO: 21.
  • the final 35 C-terminal amino acids of SEQ ID NO: 21 can usefully be omitted.
  • the first 40 N-terminal amino acids of SEQ ID NO: 21 can usefully be omitted. Other fragments omit one or more protein domains.
  • Reference 59 discloses antigens which usefully include epitopes from both IsdB and IsdH. isdl
  • the 'isdl' antigen is annotated as 'heme-degrading monooxygenase Isdl'.
  • isdl is SAOUHSC OO 130 and has amino acid sequence SEQ ID NO: 22 (GI: 88193943).
  • Useful isdl antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 22 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%,
  • SEQ ID NO: 22 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 22; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
  • V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
  • isdl proteins include variants of SEQ ID NO: 22.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 22.
  • Other preferred fragments lack one or more amino acids
  • lukD The 'lukD' antigen is annotated as 'leukotoxin LukD'.
  • lukD is SAOUHSC 01954 and has amino acid sequence SEQ ID NO: 23 (GI:88195647). hi the Newman strain it is nwmn l 718 (GI: 151221930).
  • Useful lukD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 23 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 23; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 23, wherein 'n' is 7 or more (e.g.
  • lukD proteins include variants of SEQ ID NO: 23.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 23.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 23 while retaining at least one epitope of SEQ ID NO: 23.
  • the final 43 C-terminal amino acids of SEQ ID NO: 23 can usefully be omitted.
  • the first 26 N-terminal amino acids of SEQ ID NO: 23 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'lukE' antigen is annotated as 'leukotoxin LukE'.
  • NCTC 8325 strain lukE is SAOUHSCJ) 1955 and has amino acid sequence SEQ ID NO: 24 (GI: 88195648).
  • Useful lukE antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 24 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 24; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 24, wherein 'n' is 7 or more (e.g.
  • These lukE proteins include variants of SEQ ID NO: 24.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 24.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 24 while retaining at least one epitope of SEQ ID NO: 24.
  • Other fragments omit one or more protein domains.
  • lukF The 'lukF' antigen is annotated as 'Leukocidin/Hemolysin toxin family LukF'.
  • lukF is SAOUHSC 02241 and has amino acid sequence SEQ ID NO: 25 (GI:88195914).
  • Useful lukF antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 25 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 25 comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 25, wherein 'n ? is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • lukF proteins include variants of SEQ ID NO: 25.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 25.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'lukS' antigen is annotated as 'probable leukocidin S subunit LukS'.
  • lukS is SAOUHSC_02243 and has amino acid sequence SEQ ID NO: 26 (GL88195915).
  • nwmn_1928 GI: 151222140.
  • Useful lukS antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 26 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 26; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 26, wherein 'n' is 7 or more (e.g.
  • These lukS proteins include variants of SEQ ID NO: 26.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 26.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 26 while retaining at least one epitope of SEQ ID NO: 26.
  • the first 22 N-terminal amino acids of SEQ ID NO: 26 can usefully be omitted.
  • Other fragments omit one or more protein domains. nuc
  • the 'nuc' antigen is annotated as 'thermonuclease precursor'.
  • NCTC 8325 strain nuc is
  • SAOUHSC 01316 has amino acid sequence SEQ ID NO: 27 (GI:88195046).
  • Useful nuc antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 27 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 27; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 27, wherein 'n' is 7 or more (e.g.
  • nuc proteins include variants of SEQ ID NO: 27.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 27.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 27 while retaining at least one epitope of SEQ ID NO: 27.
  • the final 39 C-terminal amino acids of SEQ ID NO: 27 can usefully be omitted.
  • the first 19 N-terminal amino acids of SEQ ID NO: 27 can usefully be omitted.
  • Other fragments omit one or more protein domains. sasA
  • the 'sasA' antigen is annotated as 'SasA'.
  • sasA is SAOUHSC 02990 and has amino acid sequence SEQ ID NO: 28 (GI:88196609).
  • Useful sasA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 28 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 28; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 28, wherein 'n' is 7 or more (e.g.
  • sasA proteins include variants of SEQ ID NO: 28.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 28.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 28 while retaining at least one epitope of SEQ ID NO: 28.
  • the final 43 C-terminal amino acids of SEQ ID NO: 28 can usefully be omitted.
  • the first 90 N-terminal amino acids of SEQ ID NO: 28 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'sasB' antigen is annotated as 'frntB protein; SasB'. hi the NCTC 8325 strain sasB is SAOUHSC 02404 and has amino acid sequence SEQ ID NO: 29 (GI: 88196065).
  • Useful sasB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 29 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 29; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 29, wherein W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sasB proteins include variants of SEQ ID NO: 29.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 29.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 29 while retaining at least one epitope of SEQ ID NO: 29.
  • the final 39 C-terminal amino acids of SEQ ID NO: 29 can usefully be omitted.
  • the first 38 N-terminal amino acids of SEQ ID NO: 29 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'sasC antigen is annotated as 'Mrp protein; SasC.
  • sasC is SAOUHSC 01873 and has amino acid sequence SEQ ID NO: 30 (GI:88195570).
  • Useful sasC antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 30 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 30; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 30, wherein V is 7 or more (e.g.
  • These sasC proteins include variants of SEQ ID NO: 30.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 30.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 30 while retaining at least one epitope of SEQ ID NO: 30.
  • the final 36 C-terminal amino acids of SEQ ID NO: 30 can usefully be omitted.
  • the first 37 N-terminal amino acids of SEQ ID NO: 30 can usefully be omitted.
  • Other fragments omit one or more protein domains. sasD
  • the 'sasD' antigen is annotated as 'SasD protein'.
  • NCTC 8325 strain sasD is
  • SAOUHSC 00094 has amino acid sequence SEQ ID NO: 31 (GL88193909).
  • Useful sasD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 31 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 31; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 31, wherein 'n' is 7 or more (e.g.
  • sasD proteins include variants of SEQ ID NO: 31.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 31.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 31 while retaining at least one epitope of SEQ ID NO: 31.
  • the first 28 N-terminal amino acids of SEQ ID NO: 31 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'sasF antigen is annotated as 'sasF protein'.
  • sasF is SAOUHSC 02982 and has amino acid sequence SEQ ID NO: 32 (GI:88196601).
  • Useful sasF antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
  • ED NO: 32 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 32; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 32, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • These sasF proteins include variants of SEQ ID NO: 32.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 32.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 32 while retaining at least one epitope of SEQ ID NO: 32.
  • the final 39 C-terminal amino acids of SEQ ID NO: 32 can usefully be omitted.
  • the first 37 N-terminal amino acids of SEQ ID NO: 32 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'sdrC antigen is annotated as 'sdrC protein 1 , hi the NCTC 8325 strain sdrC is SAOUHSC 00544 and has amino acid sequence SEQ ID NO: 33 (GI: 88194324).
  • Useful sdrC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 33 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 33; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 33, wherein 'n' is 7 or more (e.g.
  • These sdrC proteins include variants of SEQ ID NO: 33.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 33.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 33 while retaining at least one epitope of SEQ ID NO: 33.
  • the final 38 C-terminal amino acids of SEQ ID NO: 33 can usefully be omitted.
  • the first 50 N-terminal amino acids of SEQ ID NO: 33 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • SdrC is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc.
  • SEQ ID NO: 164 is a useful fragment of SEQ ID NO: 33 ('SdrC5i -5 i8')- This fragment includes the most exposed domain of SdrC and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. sdrD The 'sdrD' antigen is annotated as 'sdrD protein'. In the NCTC 8325 strain sdrD is SAOUHSC 00545 and has amino acid sequence SEQ ID NO: 34 (GI: 88194325).
  • Useful sdrD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 34 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 34; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 34, wherein 'n' is 7 or more (e.g.
  • These sdrD proteins include variants of SEQ ID NO: 34.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 34.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 34 while retaining at least one epitope of SEQ ID NO: 34.
  • the final 38 C-terminal amino acids of SEQ ID NO: 34 can usefully be omitted.
  • SEQ ID NO: 34 The first 52 N-terminal amino acids of SEQ ID NO: 34 can usefully be omitted. Other fragments omit one or more protein domains. SdrD is naturally a long protein and so the use of fragments is very helpful e.g. for purification, handling, fusion, expression, etc.
  • SEQ ID NO: 156 is a useful fragment of SEQ ID NO: 34 ('SdTD 53-592 '). This fragment includes the most exposed domain of SdrD and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. Another useful fragment, with the same C-terminus residue, is SdTD 394-592 (also known as SdrD-N3; SEQ ID NO: 199). Another useful fragment is SEQ ID NO: 236 (amino acids 593-1123 of SEQ ID NO: 34), referred to herein as 'SdrD Cna B'. sdrE2
  • the 'sdrE2' antigen is annotated as 'Ser-Asp rich fibrinogen/bone sialoprotein-binding protein SdrE'.
  • sdrE2 is NWMN 0525 and has amino acid sequence SEQ ID NO: 35 (GL151220737).
  • Useful sdrE2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 35 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 35 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 35, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • These sdrE2 proteins include variants of SEQ ID NO: 35.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 35.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 35 while retaining at least one epitope of SEQ YD NO: 35.
  • the final 38 C- terminal amino acids of SEQ ID NO: 35 can usefully be omitted.
  • the first 52 N-terminal amino acids of SEQ ID NO: 35 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • SdrE2 is naturally a long protein and so the use of fragments is very helpful e.g. for purification, handling, fusion, expression, etc.
  • SEQ ID NO: 155 is a useful fragment of SEQ ID NO: 35 ('SdrE 53-632 '). This fragment includes the most exposed domain of SdrE2 and is more easily used at an industrial scale. It also reduces the antigen' s similarity with human proteins. spa
  • the 'spa' antigen is annotated as 'protein A' or 'SpA'.
  • spa is SAOUHSC 00069 and has amino acid sequence SEQ ID NO: 36 (GI:88193885).
  • nwmn_0055 GI: 151220267.
  • All S.aureus strains express the structural gene for spa, a well characterized virulence factor whose cell wall-anchored surface protein product has five highly homologous immunoglobulin binding domains designated E, D, A, B, and C [60]. These domains display -80% identity at the amino acid level, are 56 to 61 residues in length, and are organized as tandem repeats [61].
  • SpA is synthesized as a precursor protein with an N-terminal signal peptide and a C-terminal sorting signal [62,63].
  • Cell wall-anchored spa is displayed in great abundance on the staphylococcal surface [64,65].
  • Each of its immunoglobulin binding domains is composed of anti- parallel ⁇ -helices that assemble into a three helix bundle and can bind the Fc domain of immunoglobulin G (IgG) [66,67], the VH3 heavy chain (Fab) of IgM ⁇ i.e. the B cell receptor) [68], the von Willebrand factor at its Al domain [69] and/or the TNF- ⁇ receptor I (TNFRI) [70], which is displayed on surfaces of airway epithelia.
  • IgG immunoglobulin G
  • Fab VH3 heavy chain
  • TNFRI TNF- ⁇ receptor I
  • Useful spa antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 36 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 36; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 36, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • These spa proteins include variants of SEQ ID NO: 36.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 36.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 36 while retaining at least one epitope of SEQ ID NO: 36.
  • the final 35 C-terminal amino acids of SEQ ID NO: 36 can usefully be omitted.
  • the first 36 N-terminal amino acids of SEQ ID NO: 36 can usefully be omitted.
  • Other fragments omit one or more protein domains. Reference 71 suggests that individual IgG-binding domains might be useful immunogens, alone or in combination.
  • SEQ ID NO: 162 is a useful fragment of SEQ ID NO: 36 ('Spa 37-325 '). This fragment contains all the five SpA Ig-binding domains (which are naturally arranged from N- to C-terminus in the order E, D, A, B, C) and includes the most exposed domain of SpA. It also reduces the antigen's similarity with human proteins. Other useful fragments may omit 1, 2, 3 or 4 of the natural A, B, C, D and/or E domains to prevent the excessive B cell expansion and then apoptosis which might occur if spa functions as a B cell superantigen.
  • other useful fragments may include only 1, 2, 3 or 4 of the natural A, B, C, D and/or E domains e.g. comprise only the SpA(A) domain but not B to E, or comprise only the SpA(D) domain but not A, B, C or E, etc.
  • a spa antigen useful with the invention may include 1, 2, 3, 4 or 5 IgG-binding domains, but ideally has 4 or fewer If an antigen includes only one type of spa domain (e.g. only the Spa(A) or SpA(D) domain), it may include more than one copy of this domain e.g. multiple SpA(D) domains in a single polypeptide chain.
  • An individual domain within the antigen may be mutated at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids relative to SEQ ID NO: 36 (e.g. see ref. 71, disclosing mutations at residues 3 and/or 24 of domain D, at residue 46 and/or 53 of domain A, etc.). Such mutants should not remove the antigen's ability to elicit an antibody that recognises SEQ ID NO: 36, but may remove the antigen's binding to IgG and/or other human proteins (such as human blood proteins).
  • a spa antigen includes a substitution at (a) one or more amino acid substitution in an IgG Fc binding sub-domain of SpA domain A, B, C, D and/or E that disrupts or decreases binding to IgG Fc, and (b) one or more amino acid substitution in a V H 3 binding sub-domain of SpA domain A, B, C, D, and/or E that disrupts or decreases binding to V H 3.
  • a variant SpA comprises at least or at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more variant SpA domain D peptides.
  • the 'staOOl 1 antigen is annotated as '5'-nucleotidase family protein', hi the NCTC 8325 strain staOOl is SAOUHSC 00025 and has amino acid sequence SEQ ID NO: 37 (GI:88193846). hi the Newman strain it is nwmn_0022 (GI: 151220234). It has also been referred to as AdsA and SasH and SA0024.
  • Useful staOOl antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 37 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 37; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 37, wherein 'n' is 7 or more (e.g.
  • staOOl proteins include variants of SEQ ID NO: 37.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 37.
  • Other preferred fragments lactone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 37 while retaining at least one epitope of SEQ ID NO: 37.
  • the final 34 C- te ⁇ ninal amino acids of SEQ ID NO: 37 can usefully be omitted.
  • the first 38 N-terminal amino acids of SEQ ID NO: 37 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta002
  • sta002' antigen is annotated as 'lipoprotein', hi the NCTC 8325 strain sta002 is SAOUHSC_00356 and has amino acid sequence SEQ ID NO: 38 (GL88194155). hi the Newman strain it is nwmn_0364 (GI: 151220576).
  • Useful sta002 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 38 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 38 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 38, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • sta002 proteins include variants of SEQ ID NO: 38.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 38.
  • Other preferred fragments lack one or more amino acids (e.g.
  • SEQ ID NO: 38 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 38 while retaining at least one epitope of SEQ ID NO: 38.
  • the first 18 N-terminal amino acids of SEQ ID NO: 38 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • SEQ ID NOs: 153 ('sta002i 9- i 87 ') and 154 ('sta002i 9-124 ') are two useful fragments of SEQ ID NO: 38 which reduce the antigen's similarity with human proteins. sta003
  • the 'staOO3' antigen is annotated as 'surface protein', hi the NCTC 8325 strain staOO3 is SAOUHSC 00400 and has amino acid sequence SEQ ID NO: 39 (GI: 88194195). In the Newman strain it is nwmn_0401 (GI: 151220613).
  • Useful staOO3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 39 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 39; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 39, wherein W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta003 proteins include variants of SEQ ID NO: 39.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 39.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 39 while retaining at least one epitope of SEQ ID NO: 39.
  • the first 32 N- terminal amino acids of SEQ ID NO: 39 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta004
  • the 'staOO4' antigen is annotated as 'Siderophore binding protein FatB'.
  • staOO4 is SAOUHSC_00749 and has amino acid sequence SEQ ID NO: 40 (GI: 88194514).
  • nwmn_0705 GI: 151220917.
  • Useful staOO4 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 40 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 40; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 40, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta004 proteins include variants of SEQ ID NO: 40.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 40.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 40 while retaining at least one epitope of SEQ ID NO: 40.
  • the first 18 N- terminal amino acids of SEQ ID NO: 40 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta005
  • the 'staOO5' antigen is annotated as 'superantigen-like protein'.
  • staOO5 is SAOUHSC Ol 127 and has amino acid sequence SEQ ID NO: 41 (GI: 88194870).
  • nwmn_1077 GI:151221289.
  • Useful sta005 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 41 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 41; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 41, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staOO5 proteins include variants of SEQ ID NO: 41.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 41.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 41 while retaining at least one epitope of SEQ ID NO: 41.
  • the first 18 N-terminal amino acids of SEQ ID NO: 41 can usefully be omitted.
  • Other fragments omit one or more protein domains. staOO ⁇
  • the 'staOO ⁇ ' antigen is annotated as 'ferrichrome-binding protein 1 , and has also been referred to as 'FhuD2' in the literature [72].
  • staOO ⁇ is SAOUHSC 02554 and has amino acid sequence SEQ ID NO: 42 (GL88196199).
  • nwmn_2185 is GI: 151222397.
  • Useful staOO ⁇ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 42 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 42; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 42, wherein 'n' is 7 or more (e.g.
  • staOO ⁇ proteins include variants of SEQ ID NO: 42.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 42.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 42 while retaining at least one epitope of SEQ ID NO: 42.
  • the first 17 N-terminal amino acids of SEQ ID NO: 42 can usefully be omitted (to provide SEQ ID NO: 246). Other fragments omit one or more protein domains. Mutant forms of staOO ⁇ are reported in reference 73.
  • a staOO ⁇ antigen may be lipidated e.g. with an acylated N-terminus cysteine.
  • One useful staOO ⁇ sequence is SEQ ID NO: 248, which has a Met-Ala-Ser- sequence at the N-terminus. sta007
  • the 'sta007' antigen is annotated as 'secretory antigen precursor', hi the NCTC 8325 strain sta007 is SAOUHSC 02571 and has amino acid sequence SEQ ID NO: 43 (GI:88196215). hi the Newman strain it is nwmn_2199 (GI: 151222411). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
  • Useful sta007 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 43 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 43 SEQ ID NO: 43; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 43, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta007 proteins include variants of SEQ ID NO: 43.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 43.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • the 'staOO8' antigen is annotated as 'lipoprotein', hi the NCTC 8325 strain staOO8 is SAOUHSC 02650 and has amino acid sequence SEQ ID NO: 44 (GI:88196290). hi the Newman strain it is nwmn_2270 (GI: 151222482).
  • Useful staOO8 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 44 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 44; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 44, wherein 'n' is 7 or more ⁇ e.g.
  • staOO8 proteins include variants of SEQ ID NO: 44.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 44.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ DD NO: 44 while retaining at least one epitope of SEQ ID NO: 44.
  • the first 17 N-terminal amino acids of SEQ ID NO: 44 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta.009
  • the 'sta009' antigen is annotated as 'immunoglobulin G-binding protein Sbi'.
  • sta009 is SAOUHSC 02706 and has amino acid sequence SEQ ID NO: 45 (GI:88196346).
  • nwmn_2317 GI: 151222529.
  • Useful sta009 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 45 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 45 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 45, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta009 proteins include variants of SEQ ID NO: 45.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 45.
  • Other preferred fragments lack one or more amino acids (e.g.
  • staOlO is SAOUHSC_02887 and has amino acid sequence SEQ ID NO: 46 (GI: 88196515).
  • nwmn_2469 GI: 151222681. Proteomic analysis has revealed that this protein is secreted or surface-exposed.
  • Useful staOlO antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 46 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 46; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 46, wherein 'n' is 7 or more (e.g.
  • staOlO proteins include variants of SEQ ID NO: 46.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 46.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 46 while retaining at least one epitope of SEQ ID NO: 46.
  • the first 29 N-terminal amino acids of SEQ ED NO: 46 can usefully be omitted.
  • Other fragments omit one or more protein domains. staOll
  • staOl l' antigen is annotated as 'lipoprotein'.
  • staOl l is SAOUHSC_00052 and has amino acid sequence SEQ ID NO: 47 (GL88193872).
  • Useful staOl l antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 47 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 47 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 47, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staOl l proteins include variants of SEQ ID NO: 47.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 47. Other preferred fragments lack one or more amino acids (e.g.
  • a staOl l antigen may be lipidated e.g. with an acylated N-terminus cysteine.
  • One useful staOl l sequence is SEQ ID NO: 249, which has a N-terminus methionine.
  • Variant forms of SEQ ID NO: 47 which may be used as or for preparing staOl 1 antigens include, but are not limited to, SEQ ID NOs: 213, 214 and 215 with various Ile/Val/Leu substitutions.
  • StaOl l can exist as a monomer or an oligomer, with Ca + * ions favouring oligomerisation.
  • the invention can use monomers and/or oligomers of StaOl 1.
  • staO12 The 'staO12' antigen is annotated as 'protein with leader'.
  • staO12 is SAOUHSC_00106 and has amino acid sequence SEQ ID NO: 48 (GI:88193919).
  • Useful staOl 2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 48 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 48; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 48, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO12 proteins include variants of SEQ ID NO: 48.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 48.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 48 while retaining at least one epitope of SEQ ID NO: 48.
  • the first 21 N- terminal amino acids of SEQ ID NO: 48 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO13
  • sta013' antigen is annotated as 'poly-gamnia-glutamate capsule biosynthesis protein', hi the NCTC 8325 strain staO13 is SAOUHSC 00107 and has amino acid sequence SEQ ID NO: 49 (GI: 88193920).
  • Useful staO13 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 49 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 49 SEQ ID NO: 49; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 49, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO13 proteins include variants of SEQ ID NO: 49.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 49.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • the 'staO14' antigen is annotated as 'lipoprotein'.
  • staO14 is
  • SAOUHSC_00137 has amino acid sequence SEQ ED NO: 50 (GI:88193950).
  • Useful staO14 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 50 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 50 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 50, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • staO14 proteins include variants of SEQ ID NO: 50.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 50.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 50 while retaining at least one epitope of SEQ ID NO: 50.
  • the first 17 N- terminal amino acids of SEQ ID NO: 50 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO15
  • the 'staO15' antigen is annotated as 'extracellular solute-binding protein; RGD containing lipoprotein'.
  • staO15 is SAOUHSC OO 170 and has amino acid sequence SEQ ID NO: 51 (GI: 88193980).
  • Useful staO15 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 51 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 51 SEQ ID NO: 51; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 51, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO15 proteins include variants of SEQ ID NO: 51.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 51.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'staOl ⁇ ' antigen is annotated as 'gamma-glutamyltranspeptidase'.
  • staOl ⁇ is SAOUHSC 00171 and has amino acid sequence SEQ ID NO: 52 (GI:88193981).
  • Useful staOl ⁇ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 52 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 52; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 52, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staOl ⁇ proteins include variants of SEQ ID NO: 52.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 52.
  • Other preferred fragments lact ⁇ one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 52 while retaining at least one epitope of SEQ ID NO: 52.
  • Other fragments omit one or more protein domains. sta017
  • the 'staO17' antigen is annotated as 'lipoprotein'.
  • staO17 is SAOUHSC OO 186 and has amino acid sequence SEQ ID NO: 53 (GI: 88193996).
  • Useful staO17 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 53 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 53; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 53, wherein 'n' is 7 or more (e.g.
  • staO17 proteins include variants of SEQ ID NO: 53.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 53.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 53 while retaining at least one epitope of SEQ ID NO: 53.
  • the first 17 N- terminal amino acids of SEQ ID NO: 53 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO18
  • the 'staO18' antigen is annotated as 'extracellular solute-binding protein'.
  • staO18 is SAOUHSC 00201 and has amino acid sequence SEQ DD NO: 54 (GL88194011).
  • Useful staO18 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 54 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 54; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 54, wherein 'n' is 7 or more ⁇ e.g.
  • staO18 proteins include variants of SEQ ID NO: 54.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 54.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 54 while retaining at least one epitope of SEQ ID NO: 54.
  • Other fragments omit one or more protein domains. staO19
  • the 'staO19' antigen is annotated as 'peptidoglycan hydrolase'.
  • staO19 is SAOUHSC_00248 and has amino acid sequence SEQ ID NO: 55 (GI:88194055).
  • the Newman strain it is nwmn_0210 (GI: 151220422).
  • Useful staO19 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 55 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 55; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 55, wherein 'n' is 7 or more ⁇ e.g.
  • staO19 proteins include variants of SEQ ID NO: 55.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 55.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 55 while retaining at least one epitope of SEQ ID NO: 55.
  • the first 25 N- terminal amino acids of SEQ DD NO: 55 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • Useful fragments are SEQ DD NOs: 228 and 229.
  • StaO19 does not adsorb well to aluminium hydroxide adjuvants, so StaO19 present in a composition may me unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate. staO2O
  • the 'sta020' antigen is annotated as 'exported protein'.
  • sta020 is SAOUHSC_00253 and has amino acid sequence SEQ ID NO: 56 (GI:88194059).
  • Useful sta020 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 56 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 56; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 56, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta020 proteins include variants of SEQ ID NO: 56.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 56.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 56 while retaining at least one epitope of SEQ ID NO: 56.
  • the first 30 N- terminal amino acids of SEQ ID NO: 56 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO21
  • the 'staO21' antigen is annotated as 'secretory antigen SsaA-like protein 1 .
  • staO21 is SAOUHSC 00256 and has amino acid sequence SEQ ID NO: 57 (GI: 88194062).
  • Useful staO21 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 57 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 57 amino acid sequence corresponding to SEQ ID NO: 57; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 57, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • staO21 proteins include variants of SEQ ID NO: 57.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 57.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 57 while retaining at least one epitope of SEQ ID NO: 57.
  • the first 24 N- terminal amino acids of SEQ ID NO: 57 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO22
  • the 'staO22' antigen is annotated as 'lipoprotein'.
  • staO22 is
  • SAOUHSC_00279 and has amino acid sequence SEQ ID NO: 58 (GI:88194083).
  • Useful staO22 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 58 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 58 SEQ ID NO: 58; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 58, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • staO22 proteins include variants of SEQ ID NO: 58.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 58.
  • Other preferred fragments lack one or more amino acids (e.g.
  • staO23 The 'staO23' antigen is annotated as ⁇ '-nucleotidase; lipoprotein e(P4) family 1 . In the NCTC 8325 strain staO23 is SAOUHSC 00284 and has amino acid sequence SEQ ID NO: 59 (GI:88194087).
  • Useful staO23 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 59 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 59; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 59, wherein 'n' is 7 or more (e.g.
  • staO23 proteins include variants of SEQ ID NO: 59.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 59.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 59 while retaining at least one epitope of SEQ ID NO: 59.
  • the first 31 N- terminal amino acids of SEQ ID NO: 59 can usefully be omitted. Other fragments omit one or more protein domains.
  • staO24 The 'staO24' antigen is annotated as 'lipase precursor'. In the NCTC 8325 strain staO24 is SAOUHSC 00300 and has amino acid sequence SEQ ID NO: 60 (GI:88194101).
  • Useful staO24 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 60 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 60; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 60, wherein 'n' is 7 or more (e.g.
  • staO24 proteins include variants of SEQ ID NO: 60.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 60.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 60 while retaining at least one epitope of SEQ ID NO: 60.
  • the first 37 N- terminal amino acids of SEQ ID NO: 60 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO25
  • the 'staO25' antigen is annotated as 'lipoprotein'.
  • staO25 is SAOUHSC_00362 and has amino acid sequence SEQ ID NO: 61 (GI: 88194160).
  • Useful staO25 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ED NO: 61 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 61; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 61, wherein 'n' is 7 or more (e.g.
  • staO25 proteins include variants of SEQ ID NO: 61.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 61.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 61 while retaining at least one epitope of SEQ ED NO: 61.
  • the first 19 N-terminal amino acids of SEQ ID NO: 61 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO26
  • the 'staO26' antigen is annotated as 'lipoprotein'.
  • staO26 is
  • SAOUHSC_00404 has amino acid sequence SEQ ID NO: 62 (GI:88194198).
  • Useful staO26 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 62 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 62; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 62, wherein ?
  • n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • These staO26 proteins include variants of SEQ ID NO: 62.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 62.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 62 while retaining at least one epitope of SEQ ID NO: 62.
  • the first 22 N- terminal amino acids of SEQ DD NO: 62 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO27
  • the 'staO27' antigen is annotated as 'probable lipase'.
  • staO27 is
  • SAOUHSC_00661 has amino acid sequence SEQ ID NO: 63 (GL88194426).
  • Useful staO27 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 63 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 63; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 63, wherein 'n' is 7 or more (e.g.
  • staO27 proteins include variants of SEQ ID NO: 63.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 63.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 63 while retaining at least one epitope of SEQ ID NO: 63.
  • the first 23 N- terminal amino acids of SEQ ID NO: 63 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO28
  • staO28' antigen is annotated as 'secretory antigen SsaA-like protein'.
  • staO28 is SAOUHSC 00671 and has amino acid sequence SEQ ID NO: 64 (GI:88194436).
  • nwmn_0634 GI: 151220846.
  • Useful staO28 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 64 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 64 SEQ ID NO: 64; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 64, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO28 proteins include variants of SEQ ID NO: 64.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 64. Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO29' antigen is annotated as 'ferrichrome binding protein'.
  • staO29 is
  • SAOUHSC 00754 and has amino acid sequence SEQ ID NO: 65 (GI: 88194518).
  • Useful staO29 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 65 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 65; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 65, wherein 'n' is 7 or more (e.g.
  • staO29 proteins include variants of SEQ ID NO: 65.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 65.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 65 while retaining at least one epitope of SEQ ID NO: 65.
  • the final 25 C- terminal amino acids of SEQ ID NO: 65 can usefully be omitted.
  • the first 19 N-terminal amino acids of SEQ DD NO: 65 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta030
  • staO3O antigen is annotated as 'lipoprotein'.
  • staO3O is SAOUHSC_00808 and has amino acid sequence SEQ ID NO: 66 (GI: 88194568).
  • Useful staO3O antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 66 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 66; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 66, wherein 'n' is 7 or more (e.g.
  • staO3O proteins include variants of SEQ ID NO: 66.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 66.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 66 while retaining at least one epitope of SEQ ID NO: 66.
  • the first 17 N-terminal amino acids of SEQ ID NO: 66 can usefully be omitted. Other fragments omit one or more protein domains.
  • staO31 The 'staO31' antigen is annotated as '5 -nucleotidase family protein'. In the NCTC 8325 strain staO31 is SAOUHSC 00860 and has amino acid sequence SEQ ID NO: 67 (GI:88194617).
  • Useful staO31 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 67 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 67; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 67, wherein 'n' is 7 or more (e.g.
  • staO31 proteins include variants of SEQ ID NO: 67.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 67.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 67 while retaining at least one epitope of SEQ ID NO: 67.
  • Other fragments omit one or more protein domains. staO32
  • the 'staO32' antigen is annotated as 'serine protease HtrA 1 .
  • staO32 is SAOUHSC_00958 and has amino acid sequence SEQ ID NO: 68 (GI:88194715).
  • Useful staO32 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 68 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 68 SEQ ID NO: 68
  • staO32 proteins include variants of SEQ ID NO: 68.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 68.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO33' antigen is annotated as 'cysteine protease precursor'.
  • staO33 is
  • SAOUHSC_00987 has amino acid sequence SEQ ID NO: 69 (GI: 88194744).
  • Useful staO33 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 69 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 69 SEQ ID NO: 69
  • staO33 proteins include variants of SEQ ID NO: 69.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 69.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO34' antigen is annotated as 'glutamyl endopeptidase precursor'.
  • staO34 is SAOUHSC 00988 and has amino acid sequence SEQ ID NO: 70 (GI:88194745).
  • Useful staO34 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 70 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 70 SEQ ID NO: 70; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 70, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO34 proteins include variants of SEQ ID NO: 70.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 70. Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO35' antigen is annotated as 'frnt protein'.
  • staO35 is SAOUHSC_00998 and has amino acid sequence SEQ ID NO: 71 (GI: 88194754).
  • Useful staO35 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 71 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 71; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 71, wherein 'n' is 7 or more (e.g.
  • staO35 proteins include variants of SEQ ID NO: 71.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 71.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 71 while retaining at least one epitope of SEQ ID NO: 71.
  • the first 25 N- terminal amino acids of SEQ ID NO: 71 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'staO36' antigen is annotated as 'iron-regulated protein with leader'.
  • staO36 is SAOUHSC_01084 and has amino acid sequence SEQ ID NO: 72 (GI:88194831).
  • Useful staO36 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 72 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 72; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 72, wherein 'n' is 7 or more (e.g.
  • staO36 proteins include variants of SEQ ID NO: 72.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 72.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 72 while retaining at least one epitope of SEQ ID NO: 72.
  • the final 27 C- terminal amino acids of SEQ ID NO: 72 can usefully be omitted.
  • the first 32 N-terminal amino acids of SEQ ID NO: 72 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO37
  • the 'staO37' antigen is annotated as 'iron ABC transporter; iron -binding protein IsdE'.
  • staO37 is SAOUHSC_01085 and has amino acid sequence SEQ ID NO: 73 (GI: 88194832).
  • Useful staO37 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 73 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 73 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 73, wherein V is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO37 proteins include variants of SEQ ID NO: 73.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 73.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • the 'staO38' antigen is annotated as 1 NPQTN specific sortase B'. hi the NCTC 8325 strain staO38 is
  • SAOUHSC 01088 has amino acid sequence SEQ ID NO: 74 (GI:88194835).
  • Useful staO38 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 74 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 74 SEQ ID NO: 74
  • staO38 proteins include variants of SEQ ID NO: 74.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 74.
  • Other preferred fragments lack one or more amino acids,(e.g.
  • the 'staO39' antigen is annotated as 'superantigen-like protein'.
  • staO39 is
  • SAOUHSC Ol 124 has amino acid sequence SEQ DD NO: 75 (GL88194868).
  • Useful staO39 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 75 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 75 comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 75, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO39 proteins include variants of SEQ ID NO: 75.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 75.
  • Other preferred fragments lack one or more amino acids (e.g.
  • staO4O is SAOUHSCJ1125 and has amino acid sequence SEQ ID NO: 76 (GI: 88194869). In the Newman strain it is nwmn_1076 (GI: 151221288).
  • Useful sta040 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 76 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 76; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 76, wherein 'n' is 7 or more (e.g.
  • sta040 proteins include variants of SEQ ID NO: 76.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 76.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 76 while retaining at least one epitope of SEQ ID NO: 76.
  • staO41 The first 21 N-terminal amino acids of SEQ ED NO: 76 can usefully be omitted. Other fragments omit one or more protein domains.
  • staO41 The ? sta041' antigen is annotated as 'fibronectin-binding protein A-related'. In the NCTC 8325 strain staO41 is SAOUHSC Ol 175 and has amino acid sequence SEQ ID NO: 77 (GI:88194914).
  • Useful staO41 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 77 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 77; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 77, wherein ? n' is 7 or more (e.g.
  • staO41 proteins include variants of SEQ ID NO: 77.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 77.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 77 while retaining at least one epitope of SEQ ID NO: 77.
  • Other fragments omit one or more protein domains.
  • the 'staO42' antigen is annotated as 'lipoprotein'.
  • staO42 is
  • Useful staO42 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 78 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 78; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 78, wherein 'n' is 7 or more ⁇ e.g.
  • staO42 proteins include variants of SEQ ID NO: 78.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 78.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 78 while retaining at least one epitope of SEQ ID NO: 78.
  • the first 18 N- terminal amino acids of SEQ ID NO: 78 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO43
  • the 'staO43' antigen is annotated as 'cell wall hydrolase', hi the NCTC 8325 strain staO43 is
  • SAOUHSC_01219 has amino acid sequence SEQ ID NO: 79 (GI:88194955).
  • Useful staO43 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ED NO: 79 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 79 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 79; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 79, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • staO43 proteins include variants of SEQ ID NO: 79.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 79.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 79 while retaining at least one epitope of SEQ ID NO: 79.
  • the first 38 N- terminal amino acids of SEQ ID NO: 79 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO44
  • the 'staO44' antigen is annotated as 'lipoprotein'.
  • staO44 is
  • SAOUHSC_01508 has amino acid sequence SEQ ID NO: 80 (GI:88195223).
  • Useful staO44 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 80 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 80 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 80, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO44 proteins include variants of SEQ ID NO: 80.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 80. Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO45' antigen is annotated as 'lipoprotein'.
  • staO45 is SAOUHSC_01627 and has amino acid sequence SEQ ID NO: 81 (GL88195337).
  • Useful staO45 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 81 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 81; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 81, wherein 'n' is 7 or more (e.g.
  • staO45 proteins include variants of SEQ ID NO: 81.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 81.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 81 while retaining at least one epitope of SEQ ID NO: 81.
  • the first 16 N-terminal amino acids of SEQ ID NO: 81 can usefully be omitted.
  • staO46 The 'staO46' antigen is annotated as 'Excalibur protein'.
  • staO46 is SAOUHSC_01918 and has amino acid sequence SEQ ID NO: 82 (GI:88195613).
  • Useful staO46 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 82 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 82; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 82, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO46 proteins include variants of SEQ ID NO: 82.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 82.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 82 while retaining at least one epitope of SEQ ID NO: 82.
  • the first 53 N-terminal amino acids of SEQ ID NO: 82 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO47
  • the 'staO47' antigen is annotated as 'lipoprotein'.
  • staO47 is SAOUHSC_01920 and has amino acid sequence SEQ ID NO: 83 (GL88195615).
  • Useful staO47 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 83 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 83; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 83, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO47 proteins include variants of SEQ ID NO: 83.
  • Preferred fragments of (b) comprise an epitope from SEQ ED NO: 83.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 83 while retaining at least one epitope of SEQ ID NO: 83.
  • the first 18 N-terminal amino acids of SEQ ED NO: 83 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO48
  • the 'staO48' antigen is annotated as 'intracellular serine protease'.
  • En the NCTC 8325 strain staO48 is SAOUHSC_01949 and has amino acid sequence SEQ ID NO: 84 (GI:88195642).
  • Useful staO48 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 84 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 84; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 84, wherein 'n' is 7 or more (e.g.
  • staO48 proteins include variants of SEQ ID NO: 84.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 84.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 84 while retaining at least one epitope of SEQ ED NO: 84.
  • the first 27 N- terminal amino acids of SEQ ID NO: 84 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO49
  • the 'staO49' antigen is annotated as 'protein export protein PrsA'.
  • staO49 is SAOUHSCJ) 1972 and has amino acid sequence SEQ ID NO: 85 (GI: 88195663).
  • GI: 88195663 amino acid sequence SEQ ID NO: 85
  • Newman strain it is nwmn_1733 (GL151221945).
  • Useful staO49 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 85 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 85 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 85, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO49 proteins include variants of SEQ ID NO: 85.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 85. Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO5O' antigen is annotated as 'staphopain thiol proteinase'.
  • sta050 is SAOUHSC 02127 and has amino acid sequence SEQ ID NO: 86 (GI:88195808).
  • Useful staO5O antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 86 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 86; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 86, wherein 'n' is 7 or more (e.g.
  • staO5O proteins include variants of SEQ ID NO: 86.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 86.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 86 while retaining at least one epitope of SEQ ID NO: 86.
  • the first 25 N- terminal amino acids of SEQ ID NO: 86 can usefully be omitted.
  • Other fragments omit one or more protein domains. staOSl
  • the 'staO51' antigen is annotated as 'protein with leader'.
  • staO51 is SAOUHSC 02147 and has amino acid sequence SEQ ID NO: 87 (GI: 88195827).
  • Useful staO51 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 87 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 87; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 87, wherein 'n' is 7 or more (e.g.
  • staO51 proteins include variants of SEQ ID NO: 87.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 87.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 87 while retaining at least one epitope of SEQ ID NO: 87.
  • the first 24 N- terminal amino acids of SEQ ID NO: 87 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO52
  • the 'staO52' antigen is annotated as 'ferric hydroxamate receptor 1'.
  • staO52 is SAOUHSC 02246 and has amino acid sequence SEQ ID NO: 88 (GL88195918).
  • Useful staO52 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 88 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 88; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 88, wherein 'n' is 7 or more ⁇ e.g.
  • staO52 proteins include variants of SEQ ID NO: 88.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 88.
  • Other preferred fragments lactone or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 88 while retaining at least one epitope of SEQ ID NO: 88.
  • the first 17 N- terminal amino acids of SEQ ED NO: 88 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO53
  • sta053' antigen is annotated as 'srdH family protein 1 .
  • staO53 is SAOUHSC_02257 and has amino acid sequence SEQ ID NO: 89 (GI: 88195928).
  • Useful staO53 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 89 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 89; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 89, wherein 'n' is 7 or more ⁇ e.g.
  • staO53 proteins include variants of SEQ ID NO: 89.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 89.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 89 while retaining at least one epitope of SEQ ID NO: 89.
  • the first 26 N- terminal amino acids of SEQ ID NO: 89 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO54
  • the 'staO54' antigen is annotated as 'Probable transglycosylase isaA precursor'.
  • staO54 is SAOUHSC 02333 and has amino acid sequence SEQ ID NO: 90 (GI: 88195999).
  • Useful staO54 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 90 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 90 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 90, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO54 proteins include variants of SEQ ID NO: 90.
  • Preferred fragments of (b) comprise an epitope from SEQ ED NO: 90. Other preferred fragments lack one or more amino acids (e.g.
  • the 'staO55' antigen is annotated as 'surface hydrolase'.
  • staO55 is
  • SAOUHSC_02448 has amino acid sequence SEQ LD NO: 91 (GL88196100).
  • Useful staO55 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 91 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 91; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 91, wherein 'n' is 7 or more (e.g.
  • staO55 proteins include variants of SEQ ID NO: 91.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 91.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 91 while retaining at least one epitope of SEQ ID NO: 91.
  • the first 31 N- terminal amino acids of SEQ ID NO: 91 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO56
  • sta056' antigen is annotated as 'hyaluronate lyase'.
  • staO56 is
  • SAOUHSC_02463 has amino acid sequence SEQ ID NO: 92 (GI:88196115).
  • Useful staO56 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 92 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 92; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 92, wherein 'n' is 7 or more (e.g.
  • staO56 proteins include variants of SEQ ID NO: 92.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 92.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 92 while retaining at least one epitope of SEQ ID NO: 92.
  • the first 24 N- terminal amino acids of SEQ ID NO: 92 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO57
  • the 'staO57' antigen is annotated as 'secretory antigen precursor SsaA'.
  • staO57 is SAOUHSC 02576 and has amino acid sequence SEQ ID NO: 93 (GI: 88196220).
  • nwmn_2203 GI:151222415).
  • Useful staO57 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 93 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 93; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 93, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • staO57 proteins include variants of SEQ ID NO: 93.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 93.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 93 while retaining at least one epitope of SEQ ID NO: 93.
  • the first 27 N-terminal amino acids of SEQ ID NO: 93 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO58
  • the 'staO58' antigen is annotated as 'Zn-binding lipoprotein adcA-like 1 .
  • staO58 is SAOUHSC 02690 and has amino acid sequence SEQ ID NO: 94 (GL88196330).
  • Useful staO58 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 94 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 94; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 94, wherein 'n' is 7 or more (e.g.
  • staO58 proteins include variants of SEQ ID NO: 94.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 94.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 94 while retaining at least one epitope of SEQ ID NO: 94.
  • the first 20 N- terminal amino acids of SEQ ID NO: 94 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO59
  • the 'staO59' antigen is annotated as 'gamma-hemolysin h-gamma-ii subunit'.
  • staO59 is SAOUHSC 02708 and has amino acid sequence SEQ ID NO: 95 (GI: 88196348).
  • Useful staO59 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 95 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 95; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 95, wherein 'n' is 7 or more (e.g.
  • staO59 proteins include variants of SEQ ID NO: 95.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 95.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 95 while retaining at least one epitope of SEQ ID NO: 95.
  • the first 20 N- terminal amino acids of SEQ ID NO: 95 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta060
  • the 'sta060' antigen is annotated as 'peptide ABC transporter; peptide-binding protein'.
  • NCTC NCTC
  • 8325 strain sta060 is SAOUHSC_02767 and has amino acid sequence SEQ ID NO: 96
  • Useful sta060 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 96 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 96; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 96, wherein 'n' is 7 or more (e.g.
  • sta060 proteins include variants of SEQ ID NO: 96.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 96.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 96 while retaining at least one epitope of SEQ ID NO: 96.
  • the first 20 N- terminal amino acids of SEQ ID NO: 96 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO ⁇ l
  • the 'staO61' antigen is annotated as 'protein with leader'.
  • staO ⁇ l is
  • SAOUHSC_02783 has amino acid sequence SEQ ID NO: 97 (GL88196419).
  • Useful staO ⁇ l antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 97 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 97; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 97, wherein 'n' is 7 or more (e.g.
  • staO61 proteins include variants of SEQ ID NO: 97.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 97.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 97 while retaining at least one epitope of SEQ ID NO: 97.
  • the first 21 N- terminal amino acids of SEQ ID NO: 97 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO62
  • the 'staO62' antigen is annotated as 'protein with leader'.
  • staO62 is
  • SAOUHSC 02788 and has amino acid sequence SEQ ID NO: 98 (GI: 88196424).
  • Useful staO62 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 98 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 98 SEQ ID NO: 98; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 98, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • staO62 proteins include variants of SEQ ID NO: 98.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 98.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 98 while retaining at least one epitope of SEQ ID NO: 98.
  • the first 22 N- terminal amino acids of SEQ ID NO: 98 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO63
  • the 'staO63' antigen is annotated as 'aureolysin'.
  • staO63 is
  • SAOUHSC_02971 has amino acid sequence SEQ ID NO: 99 (GL88196592).
  • Useful staO63 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 99 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 99 SEQ ID NO: 99; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 99, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
  • staO63 proteins include variants of SEQ ID NO: 99.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 99.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 99 while retaining at least one epitope of SEQ ID NO: 99.
  • the first 16 N- terminal amino acids of SEQ ED NO: 99 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • staO64 The 'staO64' antigen is annotated as 'lipase', hi the NCTC 8325 strain staO64 is SAOUHSC 03006 and has amino acid sequence SEQ ID NO: 100 (GI:88196625). hi the Newman strain it is nwmn_2569 (GI: 151222781).
  • Useful staO64 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 100 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 100; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 100, wherein 'n' is 7 or more ⁇ e.g.
  • staO64 proteins include variants of SEQ ID NO: 100.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 100.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 100 while retaining at least one epitope of SEQ ID NO: 100.
  • the first 34 N-terminal amino acids of SEQ ID NO: 100 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO65
  • the 'staO65' antigen is annotated as '1-phosphatidylinositol phosphodiesterase precursor'.
  • staO65 is SAOUHSC 00051 and has amino acid sequence SEQ ED NO: 101 (GI:88193871).
  • Useful staO65 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 101 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 101 comprising a fragment of at least 'n' consecutive amino acids of SEQ ED NO: 101, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • staO65 proteins include variants of SEQ ED NO: 101.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 101.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 101 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO66
  • the 'staO66' antigen is annotated as 'protein'.
  • staO66 is SAOUHSC 00172 and has amino acid sequence SEQ ID NO: 102 (GI:88193982).
  • Useful staO66 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 102 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 102; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 102, wherein 'n' is 7 or more ⁇ e.g.
  • staO66 proteins include variants of SEQ ID NO: 102.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 102.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 102 while retaining at least one epitope of SEQ ID NO: 102.
  • the first 21 N-terminal amino acids of SEQ ID NO: 102 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO67
  • the 'staO67' antigen is annotated as 'bacterial extracellular solute-binding protein'.
  • staO67 is SAOUHSC 00176 and has amino acid sequence SEQ ID NO: 103 (GL88193986).
  • Useful staO67 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 103 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 103 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 103, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • staO67 proteins include variants of SEQ ID NO: 103.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 103.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 103 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO68
  • the 'staO68' antigen is annotated as 'iron permease FTRl'.
  • staO68 is
  • SAOUHSC_00327 has amino acid sequence SEQ ID NO: 104 (GL88194127).
  • Useful staO68 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 104 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 104 ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 104; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 104, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • staO68 proteins include variants of SEQ ID NO: 104.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 104.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-te ⁇ ninus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 104 while retaining at least one epitope of SEQ ID NO: 104.
  • the final 20 C-terminal amino acids of SEQ ID NO: 104 can usefully be omitted.
  • the first 14 N-terminal amino acids of SEQ ID NO: 104 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta069
  • the 'staO69' antigen is annotated as 'autolysin precursor'.
  • staO69 is
  • SAOUHSC_00427 has amino acid sequence SEQ ID NO: 105 (GL88194219).
  • Useful staO69 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 105 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 105 amino acid sequence encoding SEQ ID NO: 105; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 105, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • staO69 proteins include variants of SEQ ID NO: 105.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 105.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 105 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta070
  • the 'sta070' antigen is annotated as 'immunogenic secreted precursor-like protein (truncated)'.
  • NCTC 8325 strain sta070 is SAOUHSC 00773 and has amino acid sequence SEQ ID NO: 106
  • Useful sta070 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 106 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 106; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 106, wherein 'n' is 7 or more ⁇ e.g.
  • sta070 proteins include variants of SEQ ID NO: 106.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 106.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 106 while retaining at least one epitope of SEQ ID NO: 106.
  • the first 24 N-terminal amino acids of SEQ ID NO: 106 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO71
  • the 'staO71' antigen is annotated as 'hemolysin'.
  • staO71 is SAOUHSC_00854 and has amino acid sequence SEQ ID NO: 107 (GL88194612).
  • Useful staO71 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 107 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 107; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 107, wherein 'n' is 7 or more (e.g.
  • staO71 proteins include variants of SEQ ID NO: 107.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 107.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 107 while retaining at least one epitope of SEQ ID NO: 107.
  • the first 24 N-terminal amino acids of SEQ ID NO: 107 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO72
  • sta072' antigen is annotated as 'extramembranal protein'.
  • staO72 is SAOUHSC 00872 and has amino acid sequence SEQ ID NO: 108 (GI: 88194629).
  • Useful staO72 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 108 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 108; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 108, wherein 'n' is 7 or more (e.g.
  • staO72 proteins include variants of SEQ ID NO: 108.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 108.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 108 while retaining at least one epitope of SEQ ID NO: 108.
  • the first 24 N-terminal amino acids of SEQ ID NO: 108 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO73
  • sta073' antigen is annotated as 'bifunctional autolysin precursor'.
  • staO73 is SAOUHSC 00994 and has amino acid sequence SEQ ID NO: 109 (GI: 88194750).
  • nwmn_0922 GI: 151221134. Proteomic analysis has revealed that this protein is secreted or surface-exposed.
  • Useful staO73 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 109 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 109; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 109, wherein 'n' is 7 or more ⁇ e.g.
  • staO73 proteins include variants of SEQ ID NO: 109.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 109.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 109 while retaining at least one epitope of SEQ ID NO: 109.
  • the first 24 N-terminal amino acids of SEQ ID NO: 109 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • a StaO73 antigen can usefully be included in a composition in combination with a Stal 12 [74].
  • StaO73 does not adsorb well to aluminium hydroxide adjuvants, so StaO73 present in a composition may be unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate. staO74
  • the 'staO74' antigen is annotated as 'factor essential for methicillin resistance'.
  • staO74 is SAOUHSC 01220 and has amino acid sequence SEQ ID NO: 110 (GL88194956).
  • Useful staO74 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 110 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 110; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 110, wherein V is 7 or more ⁇ e.g.
  • staO74 proteins include variants of SEQ ID NO: 110.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 110.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 110 while retaining at least one epitope of SEQ ID NO: 110.
  • Other fragments omit one or more protein domains. staO75
  • the 'staO75' antigen is annotated as 'insulysin; peptidase family Ml 6'.
  • staO75 is SAOUHSC 01256 and has amino acid sequence SEQ ID NO: 111 (GI: 88194989).
  • Useful staO75 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 111 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 111; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 111, wherein 'n' is 7 or more ⁇ e.g.
  • staO75 proteins include variants of SEQ ID NO: 111.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 111.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 111 while retaining at least one epitope of SEQ ID NO: 111.
  • Other fragments omit one or more protein domains. staO76
  • the 'staO76' antigen is annotated as 'hydrolase'.
  • staO76 is SAOUHSC 01263 and has amino acid sequence SEQ ID NO: 112 (GI:88194996).
  • Useful staO76 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 112 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 112; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 112, wherein 'n' is 7 or more ⁇ e.g.
  • staO76 proteins include variants of SEQ ID NO: 112.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 112.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 112 while retaining at least one epitope of SEQ ID NO: 112.
  • the first 24 N-terminal amino acids of SEQ ID NO: 112 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO77
  • the 'staO77' antigen is annotated as 'protein'.
  • staO77 is SAOUHSC 01317 and has amino acid sequence SEQ ID NO: 113 (GI:88195047). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
  • Useful staO77 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 113 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 113 SEQ ID NO: 113
  • staO77 proteins include variants of SEQ ID NO: 113.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 113.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 113 while retaining at least one epitope of SEQ ID NO: 113.
  • the first 20 N-terminal amino acids of SEQ ID NO: 113 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO78
  • the 'staO78' antigen is annotated as 'FtsK/SpoIIIE family protein'.
  • staO78 is SAOUHSC 01857 and has amino acid sequence SEQ ID NO: 114 (GI:88195555).
  • Useful staO78 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 114 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 114; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 114, wherein 'n' is 7 or more (e.g.
  • staO78 proteins include variants of SEQ ID NO: 114.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 114.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 114 while retaining at least one epitope of SEQ ID NO: 114.
  • Other fragments omit one or more protein domains.
  • staO79 The 'staO79' antigen is annotated as 'serine protease SpIF'.
  • staO79 is SAOUHSC_01935 and has amino acid sequence SEQ ID NO: 115 (GI:88195630).
  • Useful staO79 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 115 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 115; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 115, wherein 'n' is 7 or more (e.g.
  • staO79 proteins include variants of SEQ ID NO: 115.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 115.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 115 while retaining at least one epitope of SEQ ID NO: 115.
  • the first 36 N-terminal amino acids of SEQ ID NO: 115 can usefully be omitted.
  • Other fragments omit one or more protein domains. sta.080
  • the 'staO8O' antigen is annotated as 'serine protease SpIE'.
  • staO8O is SAOUHSC 01936 and has amino acid sequence SEQ ID NO: 116 (GI:88195631).
  • Useful staO8O antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 116 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 116; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 116, wherein 'n' is 7 or more (e.g.
  • staO8O proteins include variants of SEQ ID NO: 116.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 116.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 116 while retaining at least one epitope of SEQ ID NO: 116.
  • the first 36 N-terminal amino acids of SEQ ID NO: 116 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO81
  • the 'staO81' antigen is annotated as 'serine protease SpID (EC:3.4.21.19)'.
  • staO81 is SAOUHSC 01938 and has amino acid sequence SEQ ID NO: 170 (GI:88195633).
  • Useful staO81 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 170 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 170; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 170, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO81 proteins include variants of SEQ ID NO: 170.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 170.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 170 while retaining at least one epitope of SEQ ID NO: 170.
  • the first 36 N-terminal amino acids of SEQ ID NO: 170 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO82
  • the 'staO82' antigen is annotated as 'serine protease SpIC.
  • staO82 is SAOUHSC 01939 and has amino acid sequence SEQ ID NO: 117 (GI: 88195634).
  • Useful staO82 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 117 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 117 SEQ ID NO: 117; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 117, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO82 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 117.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 117 while retaining at least one epitope of SEQ ID NO: 117.
  • the first 36 N-terminal amino acids of SEQ ID NO: 117 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO83
  • the 'staO83' antigen is annotated as 'serine protease SpIB'.
  • staO83 is SAOUHSC 01941 and has amino acid sequence SEQ ID NO: 118 (GI:88195635).
  • Useful staO83 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 118 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 118; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 118, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • These staO83 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 118.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 118 while retaining at least one epitope of SEQ ID NO: 118.
  • the first 36 N-terminal amino acids of SEQ ID NO: 118 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO84
  • the 'staO84' antigen is annotated as 'serine protease SpIA'.
  • staO84 is SAOUHSC_01942 and has amino acid sequence SEQ ID NO: 119 (GI:88195636).
  • Useful staO84 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 119 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 119; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 119, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • These staO84 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 119.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 119 while retaining at least one epitope of SEQ ID NO: 119.
  • the first 35 N-terminal amino acids of SEQ ID NO: 119 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO85
  • sta085' antigen is annotated as 'staphylokinase precursor'.
  • staO85 is
  • SAOUHSC_02171 has amino acid sequence SEQ ID NO: 120 (GI:88195848).
  • Useful staO85 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 120 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 120 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 120, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These staO85 proteins include variants of SEQ ID NO: 120.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 120.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 120 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO86
  • sta086' antigen is annotated as 'OxaA-like protein'.
  • staO86 is
  • SAOUHSC 02327 has amino acid sequence SEQ ID NO: 121 (GI:88195993).
  • Useful staO86 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 121 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 121 amino acid sequence 121; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 121, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • staO86 proteins include variants of SEQ ID NO: 121.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 121.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 121 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO87
  • the 'staO87' antigen is annotated as 'teicoplanin resistance protein TcaA'.
  • staO87 is SAOUHSC 02635 and has amino acid sequence SEQ ID NO: 122 (GI: 88196276).
  • Useful staO87 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 122 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 122; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 122, wherein 'n' is 7 or more (e.g.
  • staO87 proteins include variants of SEQ ID NO: 122.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 122.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 122 while retaining at least one epitope of SEQ ID NO: 122.
  • Other fragments omit one or more protein domains. staO88
  • the 'staO88' antigen is annotated as 'esterase', hi the NCTC 8325 strain staO88 is SAOUHSC 02844 and has amino acid sequence SEQ ID NO: 123 (GI:88196477).
  • Useful staO88 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 123 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 123; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 123, wherein 'n' is 7 or more (e.g.
  • staO88 proteins include variants of SEQ ID NO: 123.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 123.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 123 while retaining at least one epitope of SEQ ID NO: 123.
  • the first 18 N-terminal amino acids of SEQ ID NO: 123 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO89
  • the 'staO89' antigen is annotated as 'LysM domain protein'.
  • staO89 is SAOUHSC 02855 and has amino acid sequence SEQ ID NO: 124 (GI:88196486).
  • Useful staO89 antigens can elicit an antibody (e.g. when administered to a human) that recognises
  • SEQ ID NO: 124 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
  • SEQ ID NO: 124 SEQ ID NO: 124; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 124, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • staO89 proteins include variants of SEQ ID NO: 124.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 124.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g.
  • sta090 The 'sta090' antigen is annotated as 'LysM domain protein'. In the NCTC 8325 strain sta090 is SAOUHSC_02883 and has amino acid sequence SEQ ID NO: 125 (GI:88196512).
  • Useful sta090 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 125 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 125; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 125, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • sta090 proteins include variants of SEQ ID NO: 125.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 125.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 125 while retaining at least one epitope of SEQ ID NO: 125.
  • the first 26 N-terminal amino acids of SEQ ID NO: 125 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • staO91 The 'staO91' antigen is annotated as 'lipoprotein'.
  • staO91 is SAOUHSC 00685 and has amino acid sequence SEQ ID NO: 126 (GI:88194450).
  • Useful staO91 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 126 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 126; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 126, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • staO91 proteins include variants of SEQ ID NO: 126.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 126.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 126 while retaining at least one epitope of SEQ ID NO: 126.
  • the first 15 N-terminal amino acids of SEQ ID NO: 126 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO92
  • the 'staO92' antigen is annotated as 'M23/M37 peptidase domain protein'.
  • staO92 is SAOUHSC 00174 and has amino acid sequence SEQ ID NO: 127 (GI:88193984).
  • Useful staO92 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 127 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 127; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 127, wherein 'n' is 7 or more (e.g.
  • staO92 proteins include variants of SEQ ID NO: 127.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 127.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 127 while retaining at least one epitope of SEQ ID NO: 127.
  • the first 25 N-terminal amino acids of SEQ ID NO: 127 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO93
  • the 'staO93' antigen is annotated as 'protein'.
  • staO93 is SAOUHSC O 1854 and has amino acid sequence SEQ ID NO: 128 (GI: 88195552).
  • Useful staO93 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 128 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 128; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ED NO: 128, wherein 'n' is 7 or more (e.g.
  • staO93 proteins include variants of SEQ ID NO: 128.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 128.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 128 while retaining at least one epitope of SEQ ED NO: 128.
  • Other fragments omit one or more protein domains.
  • the 'staO94' antigen is annotated as 'protein'.
  • staO94 is SAOUHSC Ol 512 and has amino acid sequence SEQ ID NO: 129 (GI: 88195226).
  • Useful staO94 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 129 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 129 SEQ ID NO: 129
  • staO94 proteins include variants of SEQ ID NO: 129.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 129.
  • Other preferred fragments lack one or more amino acids (e.g.
  • staO95 is annotated as 'superantigen-like protein'.
  • staO95 is SAOUHSC 00383 and has amino acid sequence SEQ ID NO: 130 (GI:88194180).
  • nwmn_0388 GI: 151220600.
  • Useful staO95 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 130 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 130 SEQ ID NO: 130; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 130, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • staO95 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 130.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 130 while retaining at least one epitope of SEQ ID NO: 130.
  • the first 32 N-terminal amino acids of SEQ ID NO: 130 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO96
  • the 'staO96' antigen is annotated as 'superantigen-like protein'.
  • staO96 is
  • SAOUHSC 00384 has amino acid sequence SEQ ID NO: 131 (GI:88194181).
  • Useful staO96 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 131 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 131; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 131, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • These staO96 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 131.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 131 while retaining at least one epitope of SEQ ID NO: 131.
  • the first 30 N-terminal amino acids of SEQ ID NO: 131 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO97
  • the 'staO97' antigen is annotated as 'superantigen-like protein'.
  • staO97 is SAOUHSC_00386 and has amino acid sequence SEQ ID NO: 132 (GI:88194182).
  • Useful staO97 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 132 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 132; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 132, wherein 'n' is 7 or more (e.g.
  • staO97 proteins include variants of SEQ ID NO: 132.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 132.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 132 while retaining at least one epitope of SEQ ID NO: 132.
  • the first 30 N-terminal amino acids of SEQ ID NO: 132 can usefully be omitted.
  • Other fragments omit one or more protein domains. staO98
  • the 'staO98' antigen is annotated as 'superantigen-like protein'.
  • staO98 is SAOUHSC_00389 and has amino acid sequence SEQ ID NO: 133 (GL88194184).
  • nwmn_0391 GI: 151220603.
  • Useful staO98 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 133 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 133 amino acid sequence 133; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 133, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • staO98 proteins include variants of SEQ ID NO: 133.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 133.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • sta099' antigen is annotated as 'superantigen-like protein 5'.
  • staO99 is SAOUHSC_00390 and has amino acid sequence SEQ ID NO: 134 (GL88194185).
  • Useful staO99 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 134 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 134; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 134, wherein 'n' is 7 or more ⁇ e.g.
  • staO99 proteins include variants of SEQ ID NO: 134.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 134.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 134 while retaining at least one epitope of SEQ ID NO: 134.
  • the first 30 N-terminal amino acids of SEQ ID NO: 134 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalOO
  • the 'stalOO' antigen is annotated as 'superantigen-like protein'.
  • NCTC 8325 strain stalOO is
  • SAOUHSC_00391 has amino acid sequence SEQ ID NO: 135 (GI:88194186).
  • Useful stalOO antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 135 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 135 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 135, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stalOO proteins include variants of SEQ ID NO: 135.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 135.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 135 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalOl
  • the 'stalOl' antigen is annotated as 'superantigen-like protein 7'.
  • stalOl is SAOUHSC_00392 and has amino acid sequence SEQ ID NO: 136 (GL88194187).
  • nwmn_0394 GI: 151220606.
  • Useful stalOl antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 136 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 136 SEQ ID NO: 136; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 136, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • stalOl proteins include variants of SEQ ID NO: 136.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 136.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'stalO2' antigen is annotated as 'superantigen-like protein', hi the NCTC 8325 strain stalO2 is
  • SAOUHSC_00393 has amino acid sequence SEQ ID NO: 137 (GI:88194188).
  • Useful stalO2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 137 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 137 amino acids of SEQ ID NO: 137; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 137, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stalO2 proteins include variants of SEQ ID NO: 137.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 137.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 137 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalO3
  • the 'stalO3' antigen is annotated as 'superantigen-like protein', hi the NCTC 8325 strain stalO3 is
  • SAOUHSC_00394 has amino acid sequence SEQ ID NO: 138 (GI:88194189).
  • Useful stalO3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 138 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 138 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 138, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stalO3 proteins include variants of SEQ ID NO: 138.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 138.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • stalO4 The 'stalO4' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain stalO4 is SAOUHSC 00395 and has amino acid sequence SEQ ID NO: 139 (GI:88194190).
  • Useful stalO4 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 139 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 139; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 139, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • These stalO4 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 139.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 139 while retaining at least one epitope of SEQ ID NO: 139.
  • Other fragments omit one or more protein domains. stalO5
  • the 'stalO5' antigen is annotated as 'superantigen-like protein'.
  • NCTC 8325 strain stalO5 is SAOUHSC 00399 and has amino acid sequence SEQ ID NO: 140 (GI:88194194).
  • Newman strain it is nwmn_0400 (GI: 151220612).
  • Useful stalO5 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 140 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 140; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 140, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more).
  • These stalO5 proteins include variants of SEQ ID NO:
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 140.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 140 while retaining at least one epitope of SEQ ID NO: 140.
  • the first 30 N-terminal amino acids of SEQ ID NO: 140 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • stalO ⁇ The 'stalO ⁇ ' antigen is annotated as 'hypothetical protein'.
  • stalO ⁇ is SAOUHSC Ol 115 and has amino acid sequence SEQ ID NO: 141 (GI: 88194861).
  • Useful stalO ⁇ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 141 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 141 SEQ ID NO: 141; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 141, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • stalO ⁇ proteins include variants of SEQ ID NO: 141.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 141.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'stalO7' antigen is annotated as 'hypothetical protein 1 .
  • NCTC 8325 strain stalO7 is SAOUHSC_00354 and has amino acid sequence SEQ ID NO: 177 (GI:88194153).
  • Useful stalO7 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 177 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 177; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 177, wherein 'n' is 7 or more (e.g.
  • stalO7 proteins include variants of SEQ ID NO: 177.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 177.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 177 while retaining at least one epitope of SEQ ID NO: 177.
  • the first 35 N-terminal amino acids of SEQ ID NO: 177 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalO8
  • the 'stalO8' antigen is annotated as 'hypothetical protein', hi the NCTC 8325 strain stalO8 is SAOUHSC_00717 and has amino acid sequence SEQ ID NO: 178 (GI:88194482).
  • Useful stalO8 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 178 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 178; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 178, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • stalO8 proteins include variants of SEQ ID NO: 178.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 178.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 178 while retaining at least one epitope of SEQ ID NO: 178.
  • the first 20 N-terminal amino acids of SEQ ID NO: 178 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalO9
  • stal09' antigen is annotated as "N-acetylmuramoyl-L-alanine amidase '.
  • NCTC 8325 strain stalO9 is SAOUHSC_02979 and has amino acid sequence SEQ ID NO: 179 (GI: 88196599).
  • Useful stalO9 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 179 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 179 99.5% or more to SEQ ID NO: 179; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 179, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stalO9 proteins include variants of SEQ ID NO: 179.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 179.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 179 can usefully be omitted.
  • Other fragments omit one or more protein domains. stallO
  • the 'stal lO' antigen is annotated as 'hypothetical protein'.
  • NCTC 8325 strain stal lO is
  • SAOUHSC_01039 has amino acid sequence SEQ ID NO: 180 (GI:88194791).
  • Useful stal lO antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 180 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 180 comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 180, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stal 10 proteins include variants of SEQ ID NO: 180.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 180.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 180 can usefully be omitted.
  • Other fragments omit one or more protein domains. stalll
  • the 'stal l I' antigen is annotated as 'hypothetical protein', hi the NCTC 8325 strain stal l l is SAOUHSC 01005 and has amino acid sequence SEQ ID NO: 181 (GI: 88194760).
  • Useful stal l l antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 181 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 181; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 181, wherein 'n' is 7 or more ⁇ e.g.
  • These stal l l proteins include variants of SEQ ID NO: 181.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 181.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 181 while retaining at least one epitope of SEQ ID NO: 181.
  • the first 20 N-terminal amino acids of SEQ ID NO: 181 can usefully be omitted.
  • Other fragments omit one or more protein domains. stain
  • the 'stal l2' antigen is annotated as a putative 'ABC transporter, substrate-binding protein'.
  • NCTC 8325 strain stal l2 is SAOUHSC 00634 and has amino acid sequence SEQ ID NO: 182 (GI: 88194402).
  • Useful stal l2 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 182 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g.
  • SEQ ID NO: 182 SEQ ID NO: 182; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 182, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • stal 12 proteins include variants of SEQ ID NO: 182.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 182.
  • Other preferred fragments lack one or more amino acids ⁇ e.g.
  • a Stal 12 antigen can usefully be included in a composition in combination with a StaO73 [74]. stalB
  • the 'stal 13' antigen is annotated as 'hypothetical protein'.
  • NCTC 8325 strain stal 13 is SAOUHSC_00728 and has amino acid sequence SEQ ID NO: 183 (GL88194493).
  • Useful stal l3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 183 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 183 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 183, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more).
  • stal 13 proteins include variants of SEQ ID NO: 183.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 183.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 'stal 14' antigen is annotated as 'hypothetical protein'.
  • NCTC 8325 strain stal 14 is SAOUHSC_00810 and has amino acid sequence SEQ ID NO: 184 (GL88194570).
  • Useful stal 14 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 184 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 184; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 184, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • These stal 14 proteins include variants of SEQ ID NO: 184.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 184.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 184 while retaining at least one epitope of SEQ ID NO: 184.
  • Other fragments omit one or more protein domains. stalls
  • the 'stal 15' antigen is annotated as 'hypothetical protein'.
  • NCTC 8325 strain stal 15 is
  • SAOUHSC_00817 has amino acid sequence SEQ ID NO: 185 (GI:88194576).
  • Useful stal 15 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 185 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 185; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 185, wherein 'n' is 7 or more (e.g.
  • These stall5 proteins include variants of SEQ ID NO: 185.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 185.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 185 while retaining at least one epitope of SEQ ID NO: 185.
  • the first 18 N-terminal amino acids of SEQ ID NO: 185 can usefully be omitted.
  • Other fragments omit one or more protein domains. stall ⁇
  • the 'stal 16' antigen is annotated as 'formyl peptide receptor-like 1 inhibitory protein '.
  • NCTC 8325 strain stal l ⁇ is SAOUHSCJ)1112 and has amino acid sequence SEQ ID NO: 186 (GI:88194858).
  • Useful stal l ⁇ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 186 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 186 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 186, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • These stal l ⁇ proteins include variants of SEQ ID NO: 186.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 186.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 186 can usefully be omitted.
  • Other fragments omit one or more protein domains. stall7
  • the 'stal 17' antigen is annotated as 'truncated beta-hemolysin'.
  • NCTC 8325 strain stal 17 is
  • SAOUHSC_02240 has amino acid sequence SEQ ID NO: 187 (GI:88195913).
  • Useful stal 17 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 187 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 187; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 187, wherein 'n' is 7 or more (e.g.
  • These stal 17 proteins include variants of SEQ ID NO: 187.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 187.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 187 while retaining at least one epitope of SEQ ID NO: 187.
  • Other fragments omit one or more protein domains. stall8
  • the 'stal l ⁇ ' antigen is annotated as 'cell division protein FtsZ'.
  • NCTC 8325 strain stal l ⁇ is SAOUHSCJ)1150 and has amino acid sequence SEQ ID NO: 188 (GI: 88194892).
  • Useful stal l ⁇ antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 188 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 188; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 188, wherein 'n' is 7 or more ⁇ e.g.
  • These stal 18 proteins include variants of SEQ ID NO: 188.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 188.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 188 while retaining at least one epitope of SEQ ID NO: 188.
  • Other fragments omit one or more protein domains. stal 19
  • the 'stal 19' antigen is annotated as 'thioredoxin'.
  • NCTC 8325 strain stal 19 is SAOUHSCJ)1100 and has amino acid sequence SEQ ID NO: 200 (GI: 88194846).
  • Useful stal 19 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 200 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 200; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 200, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more).
  • These stal 19 proteins include variants of SEQ ID NO: 200.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 200.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 200 while retaining at least one epitope of SEQ ID NO: 200.
  • Other fragments omit one or more protein domains.
  • stal20 The 'stal 20' antigen is annotated as ' alkyl hydroperoxide reductase subunit C. In the NCTC 8325 strain stal20 is SAOUHSC 00365 and has amino acid sequence SEQ ID NO: 201 (GI:88194163).
  • Useful stal 20 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 201 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 201; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 201, wherein V is 7 or more ⁇ e.g.
  • These stal20 proteins include variants of SEQ ID NO: 201.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 201.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 201 while retaining at least one epitope of SEQ ID NO: 201.
  • Other fragments omit one or more protein domains.
  • the 1 NW o' antigen is annotated as 'secreted von Willebrand factor-binding protein precursor'.
  • NW 6 is NWMN 0757 and has amino acid sequence SEQ ID NO: 142 (GI: 151220969).
  • Useful NW 6 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 142 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 142; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 142, wherein 'n' is 7 or more (e.g.
  • NW 6 proteins include variants of SEQ ID NO: 142.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 142.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 142 while retaining at least one epitope of SEQ ID NO: 142.
  • the first 13 N-terminal amino acids of SEQ ID NO: 142 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • NW 9 is NWMN 0958 and has amino acid sequence SEQ ID NO: 143 (GI: 151221170).
  • Useful NW 9 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 143 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 143; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 143, wherein 'n' is 7 or more (e.g.
  • NW 9 proteins include variants of SEQ ID NO: 143.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 143.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 143 while retaining at least one epitope of SEQ ID NO: 143.
  • the first 19 N-terminal amino acids of SEQ ID NO: 143 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 1 NW lO' antigen is annotated as 'fibrinogen binding-related protein'.
  • NW_10 is NWMN_1066 and has amino acid sequence SEQ ID NO: 144 (GL151221278).
  • Useful NW lO antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 144 and/or may comprise an amino acid sequence: (a) having 50% or more identity ⁇ e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 144; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 144, wherein 'n' is 7 or more ⁇ e.g.
  • NW lO proteins include variants of SEQ ID NO: 144.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 144.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 144 while retaining at least one epitope of SEQ ID NO: 144.
  • the first 20 N-terminal amino acids of SEQ ID NO: 144 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the r NW_7' antigen is annotated as 'staphylococcal complement inhibitor SCIN'.
  • NW 7 is NWMN 1876 and has amino acid sequence SEQ ID NO: 145 (GI: 151222088).
  • Useful NW 7 antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 145 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 145 99.5% or more to SEQ ID NO: 145; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 145, wherein 'n' is 7 or more ⁇ e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
  • NW 7 proteins include variants of SEQ ID NO: 145.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 145.
  • Other preferred fragments lack one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
  • N-terminal amino acids of SEQ ID NO: 145 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 1 NWJ' antigen is annotated as 'chemotaxis-inhibiting protein CHIPS'.
  • NWJ is NWMN_1877 and has amino acid sequence SEQ ID NO: 146 (GI: 151222089).
  • Useful NWJ antigens can elicit an antibody ⁇ e.g. when administered to a human) that recognises SEQ ID NO: 146 and/or may comprise an amino acid sequence: (a) having 50% or more identity
  • SEQ ID NO: 146 SEQ ID NO: 146
  • NWJ proteins include variants of SEQ ID NO: 146.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 146.
  • Other preferred fragments lack one or more amino acids (e.g.
  • the 1 NWJ' antigen is annotated as 'enterotoxin type A precursor'.
  • NWJ is NWMNJ 883 and has amino acid sequence SEQ ID NO: 147 (GI: 151222095).
  • Useful NWJ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 147 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 147; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 147, wherein 'n' is 7 or more (e.g.
  • NWJ proteins include variants of SEQ ID NO: 147.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 147.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 147 while retaining at least one epitope of SEQ ID NO: 147.
  • the first 16 N-terminal amino acids of SEQ ID NO: 147 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the TsTWJ' antigen is annotated as 'lipoprotein'.
  • NWJ is NWMN 1924 and has amino acid sequence SEQ ID NO: 148 (GI: 151222136).
  • Useful NWJ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 148 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 148; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 148, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more).
  • NWJ proteins include variants of SEQ ID NO: 148.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 148.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 148 while retaining at least one epitope of SEQ ID NO: 148.
  • the first 17 N-terminal amino acids of SEQ ID NO: 148 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • the 'NW_5' antigen is annotated as 'cell wall surface anchor family protein'.
  • NWJ is NWMN 2392 and has amino acid sequence SEQ ID NO: 149 (GI: 151222604).
  • Useful NW 5 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 149 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 149; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 149, wherein 'n' is 7 or more (e.g.
  • NWJ proteins include variants of SEQ ID NO: 149.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 149.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 149 while retaining at least one epitope of SEQ ID NO: 149.
  • the first 52 N-terminal amino acids of SEQ ID NO: 149 can usefully be omitted.
  • Other fragments omit one or more protein domains.
  • Antigens used in the invention may be present in the composition as individual separate polypeptides. Where more than one antigen is used, however, they do not have to be present as separate polypeptides. Instead, at least two (e.g. 2, 3, 4, 5, or more) antigens can be expressed as a single polypeptide chain (a 'hybrid' polypeptide).
  • Hybrid polypeptides offer two main advantages: first, a polypeptide that may be unstable or poorly expressed on its own can be assisted by adding a suitable hybrid partner that overcomes the problem; second, commercial manufacture is simplified as only one expression and purification need be employed in order to produce two polypeptides which are both antigenically useful.
  • the hybrid polypeptide may comprise two or more polypeptide sequences from the first antigen group.
  • the hybrid polypeptide may comprise one or more polypeptide sequences from the first antigen group and one or more polypeptide sequences from the second antigen group.
  • the hybrid polypeptide may comprise two or more polypeptide sequences from each of the antigens listed above, or two or more variants of the same antigen in the cases in which the sequence has partial variability across strains.
  • Hybrids consisting of amino acid sequences from two, three, four, five, six, seven, eight, nine, or ten antigens are useful, hi particular, hybrids consisting of amino acid sequences from two, three, four, or five antigens are preferred, such as two or three antigens.
  • Different hybrid polypeptides may be mixed together in a single formulation.
  • Hybrids may be combined with non-hybrid antigens selected from the first, second or third antigen groups. Within such combinations, an antigen may be present in more than one hybrid polypeptide and/or as a non- hybrid polypeptide. It is preferred, however, that an antigen is present either as a hybrid or as a non-hybrid, but not as both.
  • hybrid polypeptides can also be combined with conjugates or non-S.aureus antigens as described above.
  • Hybrid polypeptides can be represented by the formula NH 2 -A- (-X-L-J n -B-COOH, wherein: X is an amino acid sequence of a S.aureus antigen, as described above; L is an optional linker amino acid sequence; A is an optional N-terminal amino acid sequence; B is an optional C-terminal amino acid sequence; n is an integer of 2 or more ⁇ e.g. 2, 3, 4, 5, 6, etc.). Usually n is 2 or 3.
  • a -X- moiety has a leader peptide sequence in its wild-type form, this may be included or omitted in the hybrid protein.
  • the leader peptides will be deleted except for that of the -X- moiety located at the N-terminus of the hybrid protein i.e. the leader peptide of Xi will be retained, but the leader peptides of X 2 ... X n will be omitted. This is equivalent to deleting all leader peptides and using the leader peptide of Xi as moiety -A-.
  • linker amino acid sequence -L- may be present or absent.
  • the hybrid may be NH 2 -X 1 -Li-X 2 -L 2 -COOH, NH 2 -X 1 -X 2 -COOH, NH 2 -Xi-L 1 -X 2 - COOH, NH 2 -X 1 -X 2 -L 2 -COOH, etc.
  • Linker amino acid sequence(s) -L- will typically be short ⁇ e.g. 20 or fewer amino acids i.e. 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1).
  • linker amino acid sequences will be apparent to those skilled in the art.
  • a useful linker is GSGGGG (SEQ ID NO: 171) or GSGSGGGG (SEQ ID NO: 172), with the Gly-Ser dipeptide being formed from a Bam ⁇ l restriction site (or two of them, to form the SEQ ID NO: 230 tetrapeptide), thus aiding cloning and manipulation, and the (GIy) 4 tetrapeptide (SEQ ID NO: 227) being a typical poly-glycine linker.
  • Other suitable linkers, particularly for use as the final L n are ASGGGS (SEQ ID NO: 173 e.g. encoded by SEQ ID NO: 174) or a Leu-Glu dipeptide.
  • -A- is an optional N-terminal amino acid sequence. This will typically be short ⁇ e.g. 40 or fewer amino acids i.e. 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19,
  • -A- is preferably an oligopeptide ⁇ e.g. with 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) which provides a
  • N-terminus methionine e.g. Met-Ala-Ser, or a single Met residue.
  • One hybrid polypeptide of the invention may include both EsxA and EsxB antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NOs: 151 ('EsxAB'; encoded by SEQ ID NO: 169) and 152 ('EsxBA') are examples of such hybrids, both having hexapeptide linkers ASGGGS (SEQ ID NO: 173).
  • Another 'EsxAB' hybrid comprises SEQ ID NO: 241, which may be provided with a N-terminus methionine (e.g. SEQ ID NO: 250).
  • Another hybrid polypeptide of the invention may include both SdrD and SdrE antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 168 ('SdrED') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include both CIfB and SdrD antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 202 ('ClfB-SdrD') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • SEQ ID NO: 203 ('SdrD-ClfB') is another example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • SEQ ID NO: 211 ('ClfB-N3-sdrD-N3') is another example of such a hybrid, where the N3 fragments of CIfB and SdrD are joined by hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include both IsdA and EsxA antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 204 ('IsdA-EsxA') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • SEQ ID NO: 209 ('isdA40-184-esxA') is another example of such a hybrid, in which ISdA 4O-I84 is joined to EsxA via linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include both IsdA and sta006 antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 221 ('isdA40-184-sta006') is an example of such a hybrid, in which ISdA 40-I84 is joined to Sta006 via hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include both HIa and sta006 antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 222 ( ⁇ laH35L-staOO6') is an example of such a hybrid, in which a H35L mutant of HIa is joined to Sta006 via hexapeptide linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include both HIa and Emp antigens. These may be in either order, N- to C- terminus.
  • SEQ ID NO: 205 ('HlaH35L-Emp') is an example of such a hybrid, in which a H35L mutant HIa is joined to Emp via linker ASGGGS (SEQ ID NO: 173).
  • SEQ ID NO: 206 ('Hla27-76-Emp') is another example of such a hybrid, in which a HIa fragment is joined to Emp via linker ASGGGS (SEQ ID NO: 173);
  • SEQ ID NO: 207 is a H35L mutant of SEQ ID NO: 206.
  • SEQ ID NO: 208 ('HlaPSGS-Emp') is another example of such a hybrid, in which a HIa mutant is joined to Emp via linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include IsdA and EsxA and EsxB antigens. These may be in any order, N- to C- terminus.
  • SEQ ID NO: 210 ('isdA40-184-esxAB') is an example of such a triple hybrid, in which ISdA 40-I84 is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).
  • the EsxAB already includes the same linker, so SEQ ID NO: 210 includes two of these linkers.
  • SEQ ID NO: 212 ('IsdA-esxAB') is another example of such a triple hybrid, in which IsdA is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).
  • Another hybrid polypeptide of the invention may include HIa and EsxA and EsxB antigens. These may be in any order, N- to C- terminus.
  • SEQ ID NO: 220 ('HlaH35L-esxAB') is an example of such a triple hybrid, in which a H35L mutant of HIa is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).
  • the EsxAB already includes the same linker, so SEQ ID NO: 220 includes two of these linkers.
  • hybrid polypeptide including HIa and EsxA and EsxB antigens is SEQ ID NO: 237 ('HlaH35L-esxAB' as used in the examples), in which a H35L mutant of HIa is joined to EsxA via linker APTARG (SEQ ID NO: 239) to replace its N-terminus, then to EsxB via linker ASGGGS (SEQ ID NO: 173) to replace its N-terminus.
  • This hybrid can be provided with a suitable N-terminal sequence such as SEQ ID NO: 240.
  • Another hybrid polypeptide of the invention may include sta006 and EsxA and EsxB antigens. These may be in any order, N- to C- terminus.
  • SEQ ID NO: 223 ('sta006-esxAB') is an example of such a triple hybrid, in which sta006 is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).
  • the EsxAB already includes the same linker, so SEQ ID NO: 223 includes two of these linkers.
  • hybrid polypeptide including sta006 and EsxA and EsxB antigens is SEQ ID NO: 238 ('staOO ⁇ - esxAB' as used in the examples), in which a staOO ⁇ is joined to EsxA via linker APTARG (SEQ ID NO: 239) to replace its N-terminus, then to EsxB via linker ASGGGS (SEQ ID NO: 173) to replace its N-terminus.
  • This hybrid can be provided with a suitable N-terminal sequence such as SEQ ID NO: 240.
  • these hybrid polypeptides can elicit an antibody (e.g. when administered to a human) that recognise each of the wild-type staphylococcal proteins (e.g.
  • hybrid e.g. which recognise both wild-type EsxA and wild-type EsxB, or which recognise both wild-type SdrD and wild-type SdrE, or which recognise both wild-type SdrD and wild-type CIfB, or which recognise both wild-type IsdA and wild-type EsxA, or which recognise both wild-type IsdA and wild-type staOO ⁇ , or which recognise both wild-type HIa and wild-type staOO ⁇ , or which recognise both wild-type HIa and wild-type Emp, or which recognise wild-type IsdA and wild-type EsxA and wild-type EsxB, or which recognise wild-type HIa and wild-type EsxA and wild-type EsxB, or which recognise wild-type sta006 and wild-type EsxA and wild-type EsxB.
  • Polypeptides used with the invention can take various forms (e.g. native, fusions, glycosylated, non-glycosylated, lipidated, non-lipidated, phosphorylated, non-phosphorylated, myristoylated, non-myristoylated, monomelic, multimeric, particulate, denatured, etc.).
  • Polypeptides used with the invention can be prepared by various means (e.g. recombinant expression, purification from cell culture, chemical synthesis, etc.). Recombinantly-expressed proteins are preferred, particularly for hybrid polypeptides.
  • Polypeptides used with the invention are preferably provided in purified or substantially purified form i.e. substantially free from other polypeptides (e.g. free from naturally-occurring polypeptides), particularly from other staphylococcal or host cell polypeptides, and are generally at least about 50% pure (by weight), and usually at least about 90% pure i.e. less than about 50%, and more preferably less than about 10% (e.g. 5%) of a composition is made up of other expressed polypeptides.
  • the antigens in the compositions are separated from the whole organism with which the molecule is expressed.
  • Polypeptides used with the invention are preferably staphylococcal polypeptides.
  • polypeptide refers to amino acid polymers of any length.
  • the polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids.
  • the terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component.
  • polypeptides containing one or more analogs of an amino acid including, for example, unnatural amino acids, etc.
  • Polypeptides can occur as single chains or associated chains.
  • the invention provides polypeptides comprising a sequence -P-Q- or -Q-P-, wherein: -P- is an amino acid sequence as defined above and -Q- is not a sequence as defined above i.e. the invention provides fusion proteins.
  • -P- is an amino acid sequence as defined above
  • -Q- is not a sequence as defined above i.e. the invention provides fusion proteins.
  • the N-terminus codon of -P- is not ATG, but this codon is not present at the N-terminus of a polypeptide, it will be translated as the standard amino acid for that codon rather than as a Met. Where this codon is at the N-terminus of a polypeptide, however, it will be translated as Met.
  • the invention also provides a process for producing a polypeptide of the invention, comprising the step of culturing a host cell transformed with nucleic acid of the invention under conditions which induce polypeptide expression.
  • expression of the polypeptides of the invention may take place in a Staphylococcus, the invention will usually use a heterologous host for expression (recombinant expression).
  • the heterologous host may be prokaryotic (e.g. a bacterium) or eukaryotic. It may be E.coli, but other suitable hosts include Bacillus subtilis, Vibrio cholerae, Salmonella typhi, Salmonella typhimurium, Neisseria lactamica, Neisseria cinerea, Mycobacteria (e.g. M.tuberculosis), yeasts, etc.
  • the invention provides a process for producing a polypeptide of the invention, comprising the step of synthesising at least part of the polypeptide by chemical means.
  • the invention also provides nucleic acid encoding polypeptides and hybrid polypeptides of the invention. It also provides nucleic acid comprising a nucleotide sequence that encodes one or more polypeptides or hybrid polypeptides of the invention.
  • the invention also provides nucleic acid comprising nucleotide sequences having sequence identity to such nucleotide sequences. Identity between sequences is preferably determined by the Smith- Waterman homology search algorithm as described above. Such nucleic acids include those using alternative codons to encode the same amino acid.
  • the invention also provides nucleic acid which can hybridize to these nucleic acids.
  • Hybridization reactions can be performed under conditions of different "stringency”. Conditions that increase stringency of a hybridization reaction of widely known and published in the art (e.g. page 7.52 of reference 276).
  • Examples of relevant conditions include (in order of increasing stringency): incubation temperatures of 25°C, 37°C, 50°C, 55°C and 68°C; buffer concentrations of 10 x SSC, 6 x SSC, 1 x SSC, 0.1 x SSC (where SSC is 0.15 M NaCl and 15 mM citrate buffer) and their equivalents using other buffer systems; formamide concentrations of 0%, 25%, 50%, and 75%; incubation times from 5 minutes to 24 hours; 1, 2, or more washing steps; wash incubation times of 1, 2, or 15 minutes; and wash solutions of 6 x SSC, 1 x SSC, 0.1 x SSC, or de-ionized water.
  • Hybridization techniques and their optimization are well known in the art (e.g. see refs 75, 76, 276, 278, etc.].
  • nucleic acid of the invention hybridizes to a target under low stringency conditions; in other embodiments it hybridizes under intermediate stringency conditions; in preferred embodiments, it hybridizes under high stringency conditions.
  • An exemplary set of low stringency hybridization conditions is 50°C and 10 x SSC.
  • An exemplary set of intermediate stringency hybridization conditions is 55°C and 1 x SSC.
  • An exemplary set of high stringency hybridization conditions is 68°C and 0.1 x SSC.
  • the invention includes nucleic acid comprising sequences complementary to these sequences (e.g. for antisense or probing, or for use as primers). Nucleic acids of the invention can be used in hybridisation reactions (e.g.
  • Northern or Southern blots or in nucleic acid microarrays or 'gene chips'
  • amplification reactions e.g. PCR, SDA, SSSR, LCR, TMA, NASBA, etc.
  • other nucleic acid techniques e.g. PCR, SDA, SSSR, LCR, TMA, NASBA, etc.
  • Nucleic acid according to the invention can take various forms (e.g. single-stranded, double-stranded, vectors, primers, probes, labelled etc.). Nucleic acids of the invention may be circular or branched, but will generally be linear. Unless otherwise specified or required, any embodiment of the invention that utilizes a nucleic acid may utilize both the double-stranded form and each of two complementary single-stranded forms which make up the double-stranded form. Primers and probes are generally single-stranded, as are antisense nucleic acids. Nucleic acids of the invention are preferably provided in purified or substantially purified form i.e. substantially free from other nucleic acids (e.g.
  • Nucleic acids of the invention are preferably staphylococcal nucleic acids.
  • Nucleic acids of the invention may be prepared in many ways e.g. by chemical synthesis (e.g. phosphoramidite synthesis of DNA) in whole or in part, by digesting longer nucleic acids using nucleases (e.g. restriction enzymes), by joining shorter nucleic acids or nucleotides (e.g. using ligases or polymerases), from genomic or cDNA libraries, etc.
  • Nucleic acid of the invention may be attached to a solid support (e.g. a bead, plate, filter, film, slide, microarray support, resin, etc.). Nucleic acid of the invention may be labelled e.g. with a radioactive or fluorescent label, or a biotin label. This is particularly useful where the nucleic acid is to be used in detection techniques e.g. where the nucleic acid is a primer or as a probe.
  • a solid support e.g. a bead, plate, filter, film, slide, microarray support, resin, etc.
  • Nucleic acid of the invention may be labelled e.g. with a radioactive or fluorescent label, or a biotin label. This is particularly useful where the nucleic acid is to be used in detection techniques e.g. where the nucleic acid is a primer or as a probe.
  • nucleic acid includes in general means a polymeric form of nucleotides of any length, which contain deoxyribonucleotides, ribonucleotides, and/or their analogs. It includes DNA, RNA, DNA/RNA hybrids. It also includes DNA or RNA analogs, such as those containing modified backbones (e.g. peptide nucleic acids (PNAs) or phosphorothioates) or modified bases.
  • PNAs peptide nucleic acids
  • the invention includes mRNA, tRNA, rRNA, ribozymes, DNA, cDNA, recombinant nucleic acids, branched nucleic acids, plasmids, vectors, probes, primers, etc.
  • nucleic acid of the invention may or may not have a 5' cap.
  • Nucleic acids of the invention may be part of a vector i.e. part of a nucleic acid construct designed for transduction/transfection of one or more cell types.
  • Vectors may be, for example, "cloning vectors” which are designed for isolation, propagation and replication of inserted nucleotides, "expression vectors” which are designed for expression of a nucleotide sequence in a host cell, "viral vectors” which is designed to result in the production of a recombinant virus or virus-like particle, or “shuttle vectors", which comprise the attributes of more than one type of vector.
  • Preferred vectors are plasmids.
  • a "host cell” includes an individual cell or cell culture which can be or has been a recipient of exogenous nucleic acid.
  • Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in total DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation and/or change.
  • Host cells include cells transfected or infected in vivo or in vitro with nucleic acid of the invention. Where a nucleic acid is DNA, it will be appreciated that "U” in a RNA sequence will be replaced by "T” in the DNA. Similarly, where a nucleic acid is RNA, it will be appreciated that "T” in a DNA sequence will be replaced by "U” in the RNA.
  • complement or “complementary” when used in relation to nucleic acids refers to Watson- Crick base pairing.
  • the complement of C is G
  • the complement of G is C
  • the complement of A is T (or U)
  • the complement of T is A.
  • bases such as I (the purine inosine) e.g. to complement pyrimidines (C or T).
  • Nucleic acids of the invention can be used, for example: to produce polypeptides; as hybridization probes for the detection of nucleic acid in biological samples; to generate additional copies of the nucleic acids; to generate ribozymes or antisense oligonucleotides; as single-stranded DNA primers or probes; or as triple-strand forming oligonucleotides.
  • the invention provides a process for producing nucleic acid of the invention, wherein the nucleic acid is synthesised in part or in whole using chemical means.
  • the invention provides vectors comprising nucleotide sequences of the invention (e.g. cloning or expression vectors) and host cells transformed with such vectors.
  • Nucleic acid amplification according to the invention may be quantitative and/or real-time.
  • nucleic acids are preferably at least 7 nucleotides in length (e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300 nucleotides or longer).
  • nucleic acids are preferably at most 500 nucleotides in length (e.g.
  • 450, 400 350, 300, 250, 200, 150, 140, 130, 120, 110, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15 nucleotides or shorter).
  • Primers and probes of the invention, and other nucleic acids used for hybridization are preferably between 10 and 30 nucleotides in length (e.g. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides).
  • Antigens are defined above by reference to existing nomenclature (e.g. "CIfA”), to “sta” numbers or to “NW “ numbers.
  • Table 1 herein relates these three naming/numbering systems to existing SAOUHSC numbering and/or NWMN numbering.
  • SAOUHSC numbering refers to the genome of S.aureus strain NCTC 8325 (sequenced by Oklahoma University Health Sciences Center and disclosed in GenBank as CP000253.1; GL87201381), and individual SAOUHSC numbers are given as "locus tag” entries in the genome sequence's "features” section.
  • NWMN numbering refers to the genome of S.aureus strain Newman (isolated in 1952 from a human infection, and having robust virulence phenotype) disclosed in GenBank as AP009351.1 (GI: 150373012) and individual NWMN numbers are given as "locus tag" entries in the genome sequence's "features” section. Functional annotations for each antigen are also given in the databases.
  • Table 1 also includes the GI number for each antigen of the invention.
  • an exemplary amino acid and nucleotide sequence for any of these antigens can easily be found in public sequence databases from the NCTC 8325 and/or Newman strain, but the invention is not limited to sequences from the
  • NCTC 8325 and Newman strains Genome sequences of several other strains of S.aureus are available, including those of MRSA strains N315 and Mu50 [77], . MW2, N315, COL, MRSA252,
  • polypeptides used with the invention may, compared to the SEQ ID NO herein, include one or more ⁇ e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) amino acid substitutions, such as conservative substitutions (i.e. substitutions of one amino acid with another which has a related side chain).
  • Genetically-encoded amino acids are generally divided into four families: (1) acidic i.e. aspartate, glutamate; (2) basic i.e. lysine, arginine, histidine; (3) non-polar i.e. alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar i.e.
  • polypeptides may also include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) single amino acid deletions relative to the SEQ ID NO sequences.
  • polypeptides may also include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) insertions (e.g. each of 1, 2, 3, 4 or 5 amino acids) relative to the SEQ ID NO sequences.
  • a polypeptide used with the invention may comprise an amino acid sequence that: is identical (i.e. 100% identical) to a sequence disclosed in the sequence listing; shares sequence identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) with a sequence disclosed in the sequence listing; has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (or more) single amino acid alterations (deletions, insertions, substitutions), which may be at separate locations or may be contiguous, as compared to the sequences of (a) or (b); and when aligned with a particular sequence from the sequence listing using a pairwise alignment algorithm, each moving window of x amino acids from N-terminus to C-terminus (such that for an alignment that extends to p amino acids, where p>x, there are p-x+1 such windows) has at least xy identical aligned amino acids, where: x is selected from 20, 25, 30, 35
  • deletions or substitutions may be at the N-terminus and/or C-terminus, or may be between the two termini.
  • Truncations may involve deletion of up to 40 (or more) amino acids at the N-terminus and/or C-terminus.
  • N-terminus truncation can remove leader peptides e.g. to facilitate recombinant expression in a heterologous host.
  • C-terminus truncation can remove anchor sequences e.g. to facilitate recombinant expression in a heterologous host.
  • an antigen comprises a sequence that is not identical to a complete S. aureus sequence from the sequence listing ⁇ e.g. when it comprises a sequence listing with ⁇ 100% sequence identity thereto, or when it comprises a fragment thereof) it is preferred in each individual instance that the antigen can elicit an antibody which recognises the respective complete S. aureus sequence.
  • the invention also provides a S. aureus bacterium in which one or more of the antigens from the various antigen groups of the invention has/have been knocked out. Techniques for producing knockout bacteria are well known, and knockout S. aureus strains have been reported.
  • a knockout mutation may be situated in the coding region of the gene or may lie within its transcriptional control regions ⁇ e.g. within its promoter).
  • a knockout mutation will reduce the level of mRNA encoding the antigen to ⁇ 1% of that produced by the wild-type bacterium, preferably ⁇ 0.5%, more preferably ⁇ 0.1%, and most preferably to 0%.
  • the invention also provides a S.
  • aureus in which one or more of the antigens from the various antigen groups of the invention has a mutation which inhibits its activity.
  • the gene encoding the antigen will have a mutation that changes the encoded amino acid sequence. Mutation may involve deletion, substitution, and/or insertion, any of which may be involve one or more amino acids.
  • the invention also provides a bacterium, such as a S.aureus bacterium, which hyper-expresses an antigen of the invention.
  • the invention also provides a bacterium, such as a S.aureus bacterium, that constitutively expresses an antigen of the invention.
  • a bacterium such as a S.aureus bacterium
  • the invention also provides a meningococcus comprising a gene encoding an antigen of the invention, wherein the gene is under the control of an inducible promoter.
  • Immunogenic compositions of the invention may be useful as vaccines.
  • Vaccines according to the invention may either be prophylactic (i.e. to prevent infection) or therapeutic (i.e. to treat infection), but will typically be prophylactic.
  • compositions may thus be pharmaceutically acceptable. They will usually include components in addition to the antigens e.g. they typically include one or more pharmaceutical carrier(s) and/or excipient(s). A thorough discussion of such components is available in reference 273.
  • compositions will generally be administered to a mammal in aqueous form. Prior to administration, however, the composition may have been in a non-aqueous form. For instance, although some vaccines are manufactured in aqueous form, then filled and distributed and administered also in aqueous form, other vaccines are lyophilised during manufacture and are reconstituted into an aqueous form at the time of use. Thus a composition of the invention may be dried, such as a lyophilised formulation.
  • the composition may include preservatives such as thiomersal or 2-phenoxyethanol. It is preferred, however, that the vaccine should be substantially free from (i.e. less than 5 ⁇ g/ml) mercurial material e.g. thiomersal-free. Vaccines containing no mercury are more preferred. Preservative-free vaccines are particularly preferred.
  • a composition may include a temperature protective agent. Further details of such agents are provided below.
  • a physiological salt such as a sodium salt.
  • Sodium chloride (NaCl) is preferred, which may be present at between 1 and 20 mg/ml e.g. about 10 ⁇ 2mg/ml NaCl.
  • Other salts that may be present include potassium chloride, potassium dihydrogen phosphate, disodium phosphate dehydrate, magnesium chloride, calcium chloride, etc.
  • Compositions will generally have an osmolality of between 200 mOsm/kg and 400 m ⁇ sm/kg, preferably between 240-360 m ⁇ sm/kg, and will more preferably fall within the range of 290-310 mOsm/kg.
  • Compositions may include one or more buffers.
  • Typical buffers include: a phosphate buffer; a Tris buffer; a borate buffer; a succinate buffer; a histidine buffer (particularly with an aluminum hydroxide adjuvant); or a citrate buffer. Buffers will typically be included in the 5-2OmM range.
  • the pH of a composition will generally be between 5.0 and 8.1, and more typically between 6.0 and 8.0 e.g. 6.5 and 7.5, or between 7.0 and 7.8.
  • the composition is preferably sterile.
  • the composition is preferably non-pyrogenic e.g. containing ⁇ 1 EU (endotoxin unit, a standard measure) per dose, and preferably ⁇ 0.1 EU per dose.
  • the composition is preferably gluten free.
  • the composition may include material for a single immunisation, or may include material for multiple immunisations (i.e. a 'multidose' kit).
  • a preservative is preferred in multidose arrangements.
  • the compositions may be contained in a container having an aseptic adaptor for removal of material.
  • Human vaccines are typically administered in a dosage volume of about 0.5ml, although a half dose (i.e. about 0.25ml) may be administered to children.
  • Immunogenic compositions of the invention may also comprise one or more immunoregulatory agents.
  • one or more of the immunoregulatory agents include one or more adjuvants.
  • the adjuvants may include a THl adjuvant and/or a TH2 adjuvant, further discussed below.
  • an immunogenic composition comprising a combination of:
  • antigen(s) selected from the first, second, third and fourth antigen groups (as defined above); and (2) an adjuvant, such as an aluminium hydroxide adjuvant (for example, one or more antigens may be adsorbed to aluminium hydroxide).
  • the invention provides an immunogenic composition comprising a combination of a sta006 antigen and an adjuvant, such as an aluminium hydroxide adjuvant.
  • an immunogenic composition comprising a combination of a staOl l antigen and an adjuvant, such as an aluminium hydroxide adjuvant.
  • Adjuvants which may be used in compositions of the invention include, but are not limited to: A. Mineral-containing compositions
  • Mineral containing compositions suitable for use as adjuvants in the invention include mineral salts, such as aluminium salts and calcium salts (or mixtures thereof).
  • Calcium salts include calcium phosphate (e.g. the "CAP" particles disclosed in ref. 80).
  • Aluminum salts include hydroxides, phosphates, sulfates, etc., with the salts taking any suitable form (e.g. gel, crystalline, amorphous, etc.). Adsorption to these salts is preferred (e.g. all antigens may be adsorbed).
  • the mineral containing compositions may also be formulated as a particle of metal salt [81].
  • the adjuvants known as aluminum hydroxide and aluminum phosphate may be used. These names are conventional, but are used for convenience only, as neither is a precise description of the actual chemical compound which is present (e.g. see chapter 9 of reference 82)).
  • the invention can use any of the "hydroxide” or "phosphate” adjuvants that are in general use as adjuvants.
  • the adjuvants known as "aluminium hydroxide” are typically aluminium oxyhydroxide salts, which are usually at least partially crystalline.
  • the adjuvants known as "aluminium phosphate” are typically aluminium hydroxyphosphates, often also containing a small amount of sulfate (i.e. aluminium hydroxyphosphate sulfate). They may be obtained by precipitation, and the reaction conditions and concentrations during precipitation influence the degree of substitution of phosphate for hydroxyl in the salt.
  • a fibrous morphology (e.g. as seen in transmission electron micrographs) is typical for aluminium hydroxide adjuvants.
  • the pi of aluminium hydroxide adjuvants is typically about 11 i.e. the adjuvant itself has a positive surface charge at physiological pH.
  • Adsorptive capacities of between 1.8-2.6 mg protein per mg Al + ⁇ at pH 7.4 have been reported for aluminium hydroxide adjuvants.
  • Aluminium phosphate adjuvants generally have a PO 4 / Al molar ratio between 0.3 and 1.2, preferably between 0.8 and 1.2, and more preferably 0.95+0.1.
  • the aluminium phosphate will generally be amorphous, particularly for hydroxyphosphate salts.
  • a typical adjuvant is amorphous aluminium hydroxyphosphate with POVAl molar ratio between 0.84 and 0.92, included at 0.6mg Al 3+ /ml.
  • the aluminium phosphate will generally be particulate (e.g. plate-like morphology as seen in transmission electron micrographs). Typical diameters of the particles are in the range 0.5-20 ⁇ m (e.g. about 5-10 ⁇ m) after any antigen adsorption.
  • Adsorptive capacities of between 0.7-1.5 mg protein per mg AT + * at pH 7.4 have been reported for aluminium phosphate adjuvants.
  • the point of zero charge (PZC) of aluminium phosphate is inversely related to the degree of substitution of phosphate for hydroxyl, and this degree of substitution can vary depending on reaction conditions and concentration of reactants used for preparing the salt by precipitation.
  • Aluminium phosphates used according to the invention will generally have a PZC of between 4.0 and 7.0, more preferably between 5.0 and 6.5 e.g. about 5.7. As shown below, adsorption of S.aureus protein antigens (except IsdA, StaO19 and StaO73) to an aluminium hydroxide adjuvant is advantageous, particularly in a multi-protein combination (in which all antigens may be adsorbed).
  • a histidine buffer can usefully be included in such adjuvanted compositions.
  • Suspensions of aluminium salts used to prepare compositions of the invention may contain a buffer ⁇ e.g. a phosphate or a histidine or a Tris buffer), but this is not always necessary.
  • the suspensions are preferably sterile and pyrogen-free.
  • a suspension may include free aqueous phosphate ions e.g. present at a concentration between 1.0 and 2O mM, preferably between 5 and 15 mM, and more preferably about 10 mM.
  • the suspensions may also comprise sodium chloride.
  • the invention can use a mixture of both an aluminium hydroxide and an aluminium phosphate. In this case there may be more aluminium phosphate than hydroxide e.g. a weight ratio of at least 2:1 e.g. >5:l, >6:l, >7:l, >8:l, >9:l, efc.
  • the concentration of Al + ⁇ in a composition for administration to a patient is preferably less than 10mg/ml e.g. ⁇ 5 mg/ml, ⁇ 4 mg/ml, ⁇ 3 mg/ml, ⁇ 2 mg/ml, ⁇ 1 mg/ml, etc.
  • a preferred range is between 0.3 and lmg/ml. A maximum of 0.85mg/dose is preferred.
  • Oil emulsion compositions suitable for use as adjuvants in the invention include squalene-water emulsions, such as MF59 [Chapter 10 of ref. 82; see also ref. 83] (5% Squalene, 0.5% Tween 80, and 0.5% Span 85, formulated into submicron particles using a microfluidizer). Complete Freund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA) may also be used.
  • CFA Complete Freund's adjuvant
  • IFA incomplete Freund's adjuvant
  • oil-in-water emulsion adjuvants typically include at least one oil and at least one surfactant, with the oil(s) and surfactant(s) being biodegradable (metabolisable) and biocompatible.
  • the oil droplets in the emulsion are generally less than 5 ⁇ m in diameter, and ideally have a sub-micron diameter, with these small sizes being achieved with a microfiuidiser to provide stable emulsions. Droplets with a size less than 220nm are preferred as they can be subjected to filter sterilization.
  • the emulsion can comprise oils such as those from an animal (such as fish) or vegetable source.
  • Sources for vegetable oils include nuts, seeds and grains. Peanut oil, soybean oil, coconut oil, and olive oil, the most commonly available, exemplify the nut oils.
  • Jojoba oil can be used e.g. obtained from the jojoba bean. Seed oils include safflower oil, cottonseed oil, sunflower seed oil, sesame seed oil and the like. In the grain group, corn oil is the most readily available, but the oil of other cereal grains such as wheat, oats, rye, rice, teff, triticale and the like may also be used.
  • 6-10 carbon fatty acid esters of glycerol and 1,2-propanediol may be prepared by hydrolysis, separation and esterification of the appropriate materials starting from the nut and seed oils.
  • Fats and oils from mammalian milk are metabolizable and may therefore be used in the practice of this invention.
  • the procedures for separation, purification, saponification and other means necessary for obtaining pure oils from animal sources are well known in the art.
  • Most fish contain metabolizable oils which may be readily recovered. For example, cod liver oil, shark liver oils, and whale oil such as spermaceti exemplify several of the fish oils which may be used herein.
  • a number of branched chain oils are synthesized biochemically in 5-carbon isoprene units and are generally referred to as terpenoids.
  • Shark liver oil contains a branched, unsaturated terpenoids known as squalene, 2,6,10,15, 19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, which is particularly preferred herein.
  • Squalane the saturated analog to squalene
  • Fish oils, including squalene and squalane are readily available from commercial sources or may be obtained by methods known in the art. Other preferred oils are the tocopherols (see below). Mixtures of oils can be used.
  • Surfactants can be classified by their 'HLB' (hydrophile/lipophile balance). Preferred surfactants of the invention have a HLB of at least 10, preferably at least 15, and more preferably at least 16.
  • the invention can be used with surfactants including, but not limited to: the polyoxyethylene sorbitan esters surfactants (commonly referred to as the Tweens), especially polysorbate 20 and polysorbate 80; copolymers of ethylene oxide (EO), propylene oxide (PO), and/or butylene oxide (BO), sold under the DOWF AXTM tradename, such as linear EO/PO block copolymers; octoxynols, which can vary in the number of repeating ethoxy (oxy-l,2-ethanediyl) groups, with octoxynol-9 (Triton X-IOO, or t-octylphenoxypolyethoxyethanol) being of particular interest; (octylphenoxy)poly
  • Non-ionic surfactants are preferred.
  • Preferred surfactants for including in the emulsion are Tween 80 (polyoxyethylene sorbitan monooleate), Span 85 (sorbitan trioleate), lecithin and Triton X- 100.
  • surfactants can be used e.g. Tween 80/Span 85 mixtures.
  • a combination of a polyoxyethylene sorbitan ester such as polyoxyethylene sorbitan monooleate (Tween 80) and an octoxynol such as t-octylphenoxypolyethoxyethanol (Triton X-IOO) is also suitable.
  • Another useful combination comprises laureth 9 plus a polyoxyethylene sorbitan ester and/or an octoxynol.
  • Preferred amounts of surfactants are: polyoxyethylene sorbitan esters (such as Tween 80) 0.01 to 1%, in particular about 0.1 %; octyl- or nonylphenoxy polyoxyethanols (such as Triton X-100, or other detergents in the Triton series) 0.001 to 0.1 %, in particular 0.005 to 0.02%; polyoxyethylene ethers (such as laureth 9) 0.1 to 20 %, preferably 0.1 to 10 % and in particular 0.1 to 1 % or about 0.5%.
  • Preferred emulsion adjuvants have an average droplets size of ⁇ 1 ⁇ m e.g.
  • oil-in-water emulsion adjuvants useful with the invention include, but are not limited to:
  • a submicron emulsion of squalene, Tween 80, and Span 85 A submicron emulsion of squalene, Tween 80, and Span 85.
  • the composition of the emulsion by volume can be about 5% squalene, about 0.5% polysorbate 80 and about 0.5% Span 85. In weight terms, these ratios become 4.3% squalene, 0.5% polysorbate 80 and 0.48% Span 85.
  • This adjuvant is known as 'MF59' [84-86], as described in more detail in Chapter 10 of ref. 87 and chapter 12 of ref. 88.
  • the MF59 emulsion advantageously includes citrate ions e.g. 1OmM sodium citrate buffer.
  • An emulsion of squalene, a tocopherol, and polysorbate 80 (Tween 80).
  • the emulsion may include phosphate buffered saline. It may also include Span 85 (e.g. at 1%) and/or lecithin. These emulsions may have from 2 to 10% squalene, from 2 to 10% tocopherol and from 0.3 to
  • Tween 80 and the weight ratio of squalene:tocopherol is preferably ⁇ 1 as this provides a more stable emulsion.
  • Squalene and Tween 80 may be present volume ratio of about 5:2 or at a weight ratio of about 11:5.
  • One such emulsion can be made by dissolving Tween 80 in PBS to give a 2% solution, then mixing 90ml of this solution with a mixture of (5g of DL- ⁇ -tocopherol and 5ml squalene), then microfiuidising the mixture.
  • the resulting emulsion may have submicron oil droplets e.g. with an average diameter of between 100 and 250nm, preferably about 180nm.
  • the emulsion may also include a 3-de-O-acylated monophosphoryl lipid A (3d-MPL).
  • 3d-MPL 3-de-O-acylated monophosphoryl lipid A
  • Another useful emulsion of this type may comprise, per human dose, 0.5-10 mg squalene, 0.5-11 mg tocopherol, and 0.1-4 mg polysorbate 80 [89].
  • An emulsion of squalene, a tocopherol, and a Triton detergent e.g. Triton X-100.
  • the emulsion may also include a 3d-MPL (see below).
  • the emulsion may contain a phosphate buffer.
  • An emulsion comprising a polysorbate (e.g. polysorbate 80), a Triton detergent (e.g. Triton X-100) and a tocopherol (e.g. an ⁇ -tocopherol succinate).
  • the emulsion may include these three components at a mass ratio of about 75:11:10 (e.g. 750 ⁇ g/ml polysorbate 80, HO ⁇ g/ml
  • Triton X-100 and lOO ⁇ g/ml ⁇ -tocopherol succinate should include any contribution of these components from antigens.
  • the emulsion may also include squalene.
  • the emulsion may also include a 3d-MPL (see below).
  • the aqueous phase may contain a phosphate buffer. • An emulsion of squalane, polysorbate 80 and poloxamer 401 ("PluronicTM L121").
  • the emulsion can be formulated in phosphate buffered saline, pH 7.4.
  • This emulsion is a useful delivery vehicle for muramyl dipeptides, and has been used with threonyl-MDP in the "SAF-I" adjuvant [90] (0.05-1% Thr-MDP, 5% squalane, 2.5% Pluronic L121 and 0.2% polysorbate 80). It can also be used without the Thr-MDP, as in the "AF" adjuvant [91] (5% squalane, 1.25% Pluronic L121 and 0.2% polysorbate 80). Microfluidisation is preferred.
  • An emulsion comprising squalene, an aqueous solvent, a polyoxyethylene alkyl ether hydrophilic nonionic surfactant (e.g. polyoxyethylene (12) cetostearyl ether) and a hydrophobic nonionic surfactant (e.g. a sorbitan ester or mannide ester, such as sorbitan monoleate or 'Span 80').
  • the emulsion is preferably thermoreversible and/or has at least 90% of the oil droplets (by volume) with a size less than 200 nm [92].
  • the emulsion may also include one or more of: alditol; a cryoprotective agent (e.g. a sugar, such as dodecylmaltoside and/or sucrose); and/or an alkylpolyglycoside.
  • the emulsion may include a TLR4 agonist [93].
  • Such emulsions may be lyophilized.
  • An emulsion having from 0.5-50% of an oil, 0.1-10% of a phospholipid, and 0.05-5% of a non-ionic surfactant.
  • preferred phospholipid components are phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, phosphatidic acid, sphingomyelin and cardiolipin. Submicron droplet sizes are advantageous.
  • Additives may be included, such as QuilA saponin, cholesterol, a saponin-lipophile conjugate (such as GPI-0100, described in reference 96, produced by addition of aliphatic amine to desacylsaponin via the carboxyl group of glucuronic acid), dimethyidioctadecylammonium bromide and/or N,N-dioctadecyl-N,N-bis
  • An emulsion comprising a mineral oil, a non-ionic lipophilic ethoxylated fatty alcohol, and a non-ionic hydrophilic surfactant (e.g. an ethoxylated fatty alcohol and/or polyoxyethylene- polyoxypropylene block copolymer) [98].
  • a non-ionic lipophilic ethoxylated fatty alcohol e.g. an ethoxylated fatty alcohol and/or polyoxyethylene- polyoxypropylene block copolymer
  • An emulsion comprising a mineral oil, a non-ionic hydrophilic ethoxylated fatty alcohol, and a non-ionic lipophilic surfactant (e.g. an ethoxylated fatty alcohol and/or polyoxyethylene- polyoxypropylene block copolymer) [98].
  • an emulsion may be mixed with antigen extemporaneously, at the time of delivery, and thus the adjuvant and antigen may be kept separately in a packaged or distributed vaccine, ready for final formulation at the time of use.
  • an emulsion is mixed with antigen during manufacture, and thus the composition is packaged in a liquid adjuvanted form,.
  • the antigen will generally be in an aqueous form, such that the vaccine is finally prepared by mixing two liquids.
  • the volume ratio of the two liquids for mixing can vary (e.g. between 5: 1 and 1 :5) but is generally about 1:1. Where concentrations of components are given in the above descriptions of specific emulsions, these concentrations are typically for an undiluted composition, and the concentration after mixing with an antigen solution will thus decrease.
  • composition includes a tocopherol
  • any of the ⁇ , ⁇ , ⁇ , ⁇ , ⁇ or ⁇ tocopherols can be used, but ⁇ -tocopherols are preferred.
  • the tocopherol can take several forms e.g. different salts and/or isomers. Salts include organic salts, such as succinate, acetate, nicotinate, etc. D- ⁇ -tocopherol and
  • DL- ⁇ -tocopherol can both be used.
  • Tocopherols are advantageously included in vaccines for use in elderly patients ⁇ e.g. aged 60 years or older) because vitamin E has been reported to have a positive effect on the immune response in this patient group [99]. They also have antioxidant properties that may help to stabilize the emulsions [100].
  • a preferred ⁇ -tocopherol is DL- ⁇ -tocopherol, and the preferred salt of this tocopherol is the succinate. The succinate salt has been found to cooperate with
  • TNF-related ligands in vivo.
  • Saponin formulations may also be used as adjuvants in the invention.
  • Saponins are a heterogeneous group of sterol glycosides and triterpenoid glycosides that are found in the bark, leaves, stems, roots and even flowers of a wide range of plant species. Saponin from the bark of the Quillaia saponaria Molina tree have been widely studied as adjuvants. Saponin can also be commercially obtained from Smilax ornata (sarsaprilla), Gypsophilla paniculata (brides veil), and Saponaria officianalis (soap root).
  • Saponin adjuvant formulations include purified formulations, such as QS21, as well as lipid formulations, such as ISCOMs.
  • QS21 is marketed as StimulonTM.
  • Saponin compositions have been purified using HPLC and RP-HPLC. Specific purified fractions using these techniques have been identified, including QS7, QS 17, QS 18, QS21, QH-A, QH-B and QH-C.
  • the saponin is QS21.
  • a method of production of QS21 is disclosed in ref. 101.
  • Saponin formulations may also comprise a sterol, such as cholesterol [102].
  • ISCOMs immunostimulating complexs
  • the ISCOM typically also include a phospholipid such as phosphatidylethanolamine or phosphatidylcholine. Any known saponin can be used in ISCOMs.
  • the ISCOM includes one or more of QuilA, QHA & QHC.
  • ISCOMs are further described in refs. 102-104.
  • the ISCOMS may be devoid of additional detergent [105].
  • a review of the development of saponin based adjuvants can be found in refs. 106 & 107.
  • Virosomes and virus-like particles can also be used as adjuvants in the invention.
  • These structures generally contain one or more proteins from a virus optionally combined or formulated with a phospholipid. They are generally non-pathogenic, non-replicating and generally do not contain any of the native viral genome.
  • the viral proteins may be recombinantly produced or isolated from whole viruses.
  • viral proteins suitable for use in virosomes or VLPs include proteins derived from influenza virus (such as HA or NA), Hepatitis B virus (such as core or capsid proteins), Hepatitis E virus, measles virus, Sindbis virus, Rotavirus, Foot-and-Mouth Disease virus, Retrovirus, Norwalk virus, human Papilloma virus, HIV, RNA-phages, Q ⁇ -phage (such as coat proteins), GA- phage, fr-phage, AP205 phage, and Ty (such as retrotransposon Ty protein pi).
  • VLPs are discussed further in refs. 108-113.
  • Virosomes are discussed further in, for example, ref. 114
  • Adjuvants suitable for use in the invention include bacterial or microbial derivatives such as non-toxic derivatives of enterobacterial lipopolysaccharide (LPS), Lipid A derivatives, immunostimulatory oligonucleotides and ADP-ribosylating toxins and detoxified derivatives thereof.
  • Non-toxic derivatives of LPS include monophosphoryl lipid A (MPL) and 3-O-deacylated MPL (3dMPL).
  • 3dMPL is a mixture of 3 de-O-acylated monophosphoryl lipid A with 4, 5 or 6 acylated chains.
  • a preferred "small particle" form of 3 De-O-acylated monophosphoryl lipid A is disclosed in ref. 115.
  • Such "small particles" of 3dMPL are small enough to be sterile filtered through a 0.22 ⁇ m membrane [115].
  • Other non-toxic LPS derivatives include monophosphoryl lipid A mimics, such as aminoalkyl glucosaminide phosphate derivatives e.g. RC-529 [ 116, 117] .
  • Lipid A derivatives include derivatives of lipid A from Escherichia coli such as OM- 174.
  • OM- 174 is described for example in refs. 118 & 119.
  • Immunostimulatory oligonucleotides suitable for use as adjuvants in the invention include nucleotide sequences containing a CpG motif (a dinucleotide sequence containing an unmethylated cytosine linked by a phosphate bond to a guanosine). Double-stranded RNAs and oligonucleotides containing palindromic or poly(dG) sequences have also been shown to be immunostimulatory.
  • the CpG' s can include nucleotide modifications/analogs such as phosphorothioate modifications and can be double-stranded or single-stranded.
  • References 120, 121 and 122 disclose possible analog substitutions e.g. replacement of guanosine with 2'-deoxy-7-deazaguanosine.
  • the adjuvant effect of CpG oligonucleotides is further discussed in refs. 123-128.
  • the CpG sequence may be directed to TLR9, such as the motif GTCGTT or TTCGTT [129].
  • the CpG sequence may be specific for inducing a ThI immune response, such as a CpG-A ODN, or it may be more specific for inducing a B cell response, such a CpG-B ODN.
  • CpG-A and CpG-B ODNs are discussed in refs. 130-132.
  • the CpG is a CpG-A ODN.
  • the CpG oligonucleotide is constructed so that the 5' end is accessible for receptor recognition.
  • two CpG oligonucleotide sequences may be attached at their 3' ends to form "immunomers". See, for example, refs. 129 & 133-135.
  • CpG7909 also known as ProMuneTM (Coley Pharmaceutical Group, Inc.).
  • the immunostimulatory oligonucleotide may be pyrimidine-rich.
  • it may comprise more than one consecutive thymidine nucleotide (e.g. TTTT, as disclosed in ref. 136), and/or it may have a nucleotide composition with >25% thymidine (e.g. >35%, >40%, >50%, >60%, >80%, etc.).
  • it may comprise more than one consecutive cytosine nucleotide (e.g.
  • CCCC as disclosed in ref. 136
  • CCCC as disclosed in ref. 136
  • oligonucleotides may be free from unmethylated CpG motifs.
  • Immunostimulatory oligonucleotides will typically comprise at least 20 nucleotides. They may comprise fewer than 100 nucleotides.
  • an adjuvant used with the invention may comprise a mixture of (i) an oligonucleotide (e.g. between 15-40 nucleotides) including at least one (and preferably multiple) CpI motifs (i.e. a cytosine linked to an inosine to form a dinucleotide), and (ii) a polycationic polymer, such as an oligopeptide (e.g. between 5-20 amino acids) including at least one (and preferably multiple) Lys-Arg-Lys tripeptide sequence(s).
  • an oligonucleotide e.g. between 15-40 nucleotides
  • CpI motifs i.e. a cytosine linked to an inosine to form a dinucleotide
  • a polycationic polymer such as an oligopeptide (e.g. between 5-20 amino acids) including at least one (and preferably multiple) Lys-Arg-Lys tripeptide sequence(s).
  • the oligonucleotide may be a deoxynucleotide comprising 26-mer sequence 5'-(IC)i 3 -3' (SEQ ID NO: 175).
  • the polycationic polymer may be a peptide comprising 11-mer amino acid sequence KLKLLLLLKLK (SEQ ID NO: 176).
  • the oligonucleotide and polymer can form complexes e.g. as disclosed in references 138 & 139.
  • Bacterial ADP-ribosylating toxins and detoxified derivatives thereof may be used as adjuvants in the invention.
  • the protein is derived from E.coli (E.coli heat labile enterotoxin "LT"), cholera ("CT"), or pertussis ("PT").
  • LT E.coli heat labile enterotoxin
  • CT cholera
  • PT pertussis
  • the use of detoxified ADP-ribosylating toxins as mucosal adjuvants is described in ref. 140 and as parenteral adjuvants in ref. 141.
  • the toxin or toxoid is preferably in the form of a holotoxin, comprising both A and B subunits.
  • the A subunit contains a detoxifying mutation; preferably the B subunit is not mutated.
  • the adjuvant is a detoxified LT mutant such as LT-K63, LT-R72, and LT-Gl 92.
  • LT-K63, LT-R72, and LT-Gl 92 are used as adjuvants.
  • a useful CT mutant is or CT-E29H [150].
  • Numerical reference for amino acid substitutions is preferably based on the alignments of the A and B subunits of ADP-ribosylating toxins set forth in ref. 151, specifically incorporated herein by reference in its entirety.
  • Human immunomodulators suitable for use as adjuvants in the invention include cytokines, such as interleukins (e.g. IL-I, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12 [152], etc.) [153], interferons (e.g. interferon- ⁇ ), macrophage colony stimulating factor, and tumor necrosis factor.
  • cytokines such as interleukins (e.g. IL-I, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12 [152], etc.) [153], interferons (e.g. interferon- ⁇ ), macrophage colony stimulating factor, and tumor necrosis factor.
  • interferons e.g. interferon- ⁇
  • macrophage colony stimulating factor e.g. interferon- ⁇
  • tumor necrosis factor e.g. tumor necrosis factor.
  • a preferred immunomodulator is IL-12.
  • Bioadhesives and mucoadhesives may also be used as adjuvants in the invention.
  • Suitable bioadhesives include esterified hyaluronic acid microspheres [154] or mucoadhesives such as cross-linked derivatives of poly(acrylic acid), polyvinyl alcohol, polyvinyl pyrollidone, polysaccharides and carboxymethylcellulose. Chitosan and derivatives thereof may also be used as adjuvants in the invention [155].
  • Microparticles may also be used as adjuvants in the invention.
  • Microparticles i.e. a particle of ⁇ 100nm to ⁇ 150 ⁇ m in diameter, more preferably ⁇ 200nm to ⁇ 30 ⁇ m in diameter, and most preferably
  • ⁇ 500nm to ⁇ 10 ⁇ m in diameter formed from materials that are biodegradable and non-toxic (e.g. a poly( ⁇ -hydroxy acid), a polyhydroxybutyric acid, a polyorthoester, a polyanhydride, a polycaprolactone, etc.), with poly(lactide-co-glycolide) are preferred, optionally treated to have a negatively-charged surface (e.g. with SDS) or a positively-charged surface (e.g. with a cationic detergent, such as CTAB).
  • a negatively-charged surface e.g. with SDS
  • a positively-charged surface e.g. with a cationic detergent, such as CTAB
  • liposome formulations suitable for use as adjuvants are described in refs. 156-158.
  • Adjuvants suitable for use in the invention include polyoxyethylene ethers and polyoxyethylene esters [159]. Such formulations further include polyoxyethylene sorbitan ester surfactants in combination with an octoxynol [160] as well as polyoxyethylene alkyl ethers or ester surfactants in combination with at least one additional non-ionic surfactant such as an octoxynol [161].
  • Preferred polyoxyethylene ethers are selected from the following group: polyoxyethylene-9-lauryl ether (laureth 9), polyoxyethylene-9-steoryl ether, polyoxytheylene-8-steoryl ether, polyoxyethylene-4- lauryl ether, polyoxyethylene-35-lauryl ether, and polyoxyethylene-23-lauryl ether.
  • a phosphazene such as poly[di(carboxylatophenoxy)phosphazene] (“PCPP") as described, for example, in references 162 and 163, may be used.
  • PCPP poly[di(carboxylatophenoxy)phosphazene]
  • Muramyl peptides examples include N-acetyl- muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP), and N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(r-2'-dipahnitoyl-5/j- glycero-3 -hydroxyphosphoryloxy)-ethylamine MTP-PE) .
  • thr-MDP N-acetyl- muramyl-L-threonyl-D-isoglutamine
  • nor-MDP N-acetyl-normuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(r-2'-dipahnitoyl-5/j-
  • imidazoquinolone compounds suitable for use adjuvants in the invention include Imiquimod ("R-837”) [164,165], Resiquimod ("R-848”) [166], and their analogs; and salts thereof (e.g. the hydrochloride salts). Further details about immunostimulatory imidazoquinolines can be found in references 167 to 171.
  • Substituted ureas useful as adjuvants include compounds of formula I, II or III, or salts thereof: II III
  • ER 803058' such as 'ER 803058', 'ER 803732', 'ER 804053', ER 804058', 'ER 804059', 'ER 804442', 'ER 804680', 'ER 804764', ER 803022 or 'ER 804057' e.g.:
  • Cyclic diguanylate ('c-di-GMP'), which has been reported as a useful adjuvant for S.aureus vaccines [175].
  • a thiosemicarbazone compound such as those disclosed in reference 176. Methods of formulating, manufacturing, and screening for active compounds are also described in reference 176.
  • the thiosemicarbazones are particularly effective in the stimulation of human peripheral blood mononuclear cells for the production of cytokines, such as TNF- ⁇ .
  • a tryptanthrin compound such as those disclosed in reference 177.
  • Methods of formulating, manufacturing, and screening for active compounds are also described in reference 177.
  • the thiosemicarbazones are particularly effective in the stimulation of human peripheral blood mononuclear cells for the production of cytokines, such as TNF- ⁇ .
  • a nucleoside analog such as: (a) Isatorabine (ANA-245; 7-thia-8-oxoguanosine):
  • MIMP Methyl inosine 5 '-monophosphate
  • a polyhydroxlated pyrrolizidine compound [192] such as one having formula:
  • R is selected from the group comprising hydrogen, straight or branched, unsubstituted or substituted, saturated or unsaturated acyl, alkyl (e.g. cycloalkyl), alkenyl, alkynyl and aryl groups, or a pharmaceutically acceptable salt or derivative thereof.
  • alkyl e.g. cycloalkyl
  • alkenyl alkynyl and aryl groups
  • pharmaceutically acceptable salt or derivative thereof examples include, but are not limited to: casuarine, casuarine-6- ⁇ -D-glucopyranose, 3-epj-casuarine, 7-ep/-casuarine, 3,7-diep/-casuarine, etc.
  • a CDId ligand such as an ⁇ -glycosylceramide [193-200] (e.g.
  • ⁇ -galactosylceramide phytosphingosine-containing ⁇ -glycosylceramides, OCH, KRN7000 [(2S,3S,4R)-l-O-( ⁇ -D- galactopyranosyl)-2-(N-hexacosanoylamino)- 1 ,3,4-octadecanetriol], CRONY- 101, 3"-O- sulfo-galactosylceramide, etc.
  • the invention may also comprise combinations of one or more of the adjuvants identified above.
  • the following adjuvant compositions may be used in the invention: (1) a saponin and an oil- in-water emulsion [202]; (2) a saponin (e.g. QS21) + a non-toxic LPS derivative (e.g. 3dMPL) [203]; (3) a saponin (e.g. QS21) + a non-toxic LPS derivative (e.g. 3dMPL) + a cholesterol; (4) a saponin (e.g.
  • RibiTM adjuvant system (RAS), (Ribi Immunochem) containing 2% squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (DetoxTM); and (8) one or more mineral salts (such as an aluminum salt) + a non-toxic derivative of LPS (such as 3dMPL).
  • MPL monophosphorylipid A
  • TDM trehalose dimycolate
  • CWS cell wall skeleton
  • LPS such as 3dMPL
  • compositions of the invention may elicit both a cell mediated immune response as well as a humoral immune response. This immune response will preferably induce long lasting (e.g. neutralising) antibodies and a cell mediated immunity that can quickly respond upon exposure to pnuemococcus.
  • CD8 T cells can express a CD8 co-receptor and are commonly referred to as Cytotoxic T lymphocytes (CTLs).
  • CTLs Cytotoxic T lymphocytes
  • CD8 T cells are able to recognized or interact with antigens displayed on MHC Class I molecules.
  • CD4 T cells can express a CD4 co-receptor and are commonly referred to as T helper cells.
  • CD4 T cells are able to recognize antigenic peptides bound to MHC class II molecules. Upon interaction with a MHC class II molecule, the CD4 cells can secrete factors such as cytokines.
  • helper T cells or CD4+ cells can be further divided into two functionally distinct subsets: THl phenotype and TH2 phenotypes which differ in their cytokine and effector function.
  • Activated THl cells enhance cellular immunity (including an increase in antigen-specific CTL production) and are therefore of particular value in responding to intracellular infections.
  • Activated THl cells may secrete one or more of IL-2, IFN- ⁇ , and TNF- ⁇ .
  • a THl immune response may result in local inflammatory reactions by activating macrophages, NK (natural killer) cells, and CD8 cytotoxic T cells (CTLs).
  • a THl immune response may also act to expand the immune response by stimulating growth of B and T cells with IL-12.
  • THl stimulated B cells may secrete IgG2a.
  • Activated TH2 cells enhance antibody production and are therefore of value in responding to extracellular infections.
  • Activated TH2 cells may secrete one or more of IL-4, IL-5, IL-6, and IL-IO.
  • a TH2 immune response may result in the production of IgGl, IgE, IgA and memory B cells for future protection.
  • An enhanced immune response may include one or more of an enhanced THl immune response and a TH2 immune response.
  • a THl immune response may include one or more of an increase in CTLs, an increase in one or more of the cytokines associated with a THl immune response (such as IL-2, IFN- ⁇ , and TNF- ⁇ ), an increase in activated macrophages, an increase in NK activity, or an increase in the production of
  • the enhanced THl immune response will include an increase in IgG2a production.
  • a THl immune response may be elicited using a THl adjuvant.
  • a THl adjuvant will generally elicit increased levels of IgG2a production relative to immunization of the antigen without adjuvant.
  • THl adjuvants suitable for use in the invention may include for example saponin formulations, virosomes and virus like particles, non-toxic derivatives of enterobacterial lipopolysaccharide (LPS), immunostimulatory oligonucleotides.
  • LPS enterobacterial lipopolysaccharide
  • Immunostimulatory oligonucleotides such as oligonucleotides containing a CpG motif, are preferred THl adjuvants for use in the invention.
  • a TH2 immune response may include one or more of an increase in one or more of the cytokines associated with a TH2 immune response (such as IL-4, IL-5, IL-6 and IL-IO), or an increase in the production of IgGl, IgE, IgA and memory B cells.
  • the enhanced TH2 immune resonse will include an increase in IgGl production.
  • a TH2 immune response may be elicited using a TH2 adjuvant.
  • a TH2 adjuvant will generally elicit increased levels of IgGl production relative to immunization of the antigen without adjuvant.
  • TH2 adjuvants suitable for use in the invention include, for example, mineral containing compositions, oil-emulsions, and ADP-ribosylating toxins and detoxified derivatives thereof. Mineral containing compositions, such as aluminium salts are preferred TH2 adjuvants for use in the invention.
  • the invention includes a composition comprising a combination of a THl adjuvant and a TH2 adjuvant.
  • a composition elicits an enhanced THl and an enhanced TH2 response, i.e., an increase in the production of both IgGl and IgG2a production relative to immunization without an adjuvant.
  • the composition comprising a combination of a THl and a TH2 adjuvant elicits an increased THl and/or an increased TH2 immune response relative to immunization with a single adjuvant ⁇ i.e., relative to immunization with a THl adjuvant alone or immunization with a TH2 adjuvant alone).
  • the immune response may be one or both of a THl immune response and a TH2 response.
  • immune response provides for one or both of an enhanced THl response and an enhanced TH2 response.
  • the enhanced immune response may be one or both of a systemic and a mucosal immune response.
  • the immune response provides for one or both of an enhanced systemic and an enhanced mucosal immune response.
  • the mucosal immune response is a TH2 immune response.
  • the mucosal immune response includes an increase in the production of IgA.
  • compositions of the invention may be prepared in various forms.
  • the compositions may be prepared as injectables, either as liquid solutions or suspensions.
  • Solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection can also be prepared ⁇ e.g. a lyophilised composition or a spray-freeze dried composition).
  • the composition may be prepared for topical administration e.g. as an ointment, cream or powder.
  • the composition may be prepared for oral administration e.g. as a tablet or capsule, as a spray, or as a syrup (optionally flavoured).
  • the composition may be prepared for pulmonary administration e.g. as an inhaler, using a fine powder or a spray.
  • the composition may be prepared as a suppository or pessary.
  • the composition may be prepared for nasal, aural or ocular administration e.g. as drops.
  • the composition may be in kit form, designed such that a combined composition is reconstituted just prior to administration to a patient.
  • kits may comprise one or more antigens in liquid form and one or more lyophilised antigens.
  • kits may comprise two vials, or it may comprise one ready-filled syringe and one vial, with the contents of the syringe being used to reactivate the contents of the vial prior to injection.
  • Immunogenic compositions used as vaccines comprise an immunologically effective amount of antigen(s), as well as any other components, as needed.
  • 'immunologically effective amount' it is meant that the administration of that amount to an individual, either in a single dose or as part of a series, is effective for treatment or prevention. This amount varies depending upon the health and physical condition of the individual to be treated, age, the taxonomic group of individual to be treated (e.g. non-human primate, primate, etc.), the capacity of the individual's immune system to synthesise antibodies, the degree of protection desired, the formulation of the vaccine, the treating doctor's assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials. Where more than one antigen is included in a composition then two antigens may be present at the same dose as each other or at different doses.
  • a composition may include a temperature protective agent, and this component may be particularly useful in adjuvanted compositions (particularly those containing a mineral adjuvant, such as an aluminium salt).
  • a liquid temperature protective agent may be added to an aqueous vaccine composition to lower its freezing point e.g. to reduce the freezing point to below 0°C.
  • the temperature protective agent also permits freezing of the composition while protecting mineral salt adjuvants against agglomeration or sedimentation after freezing and thawing, and may also protect the composition at elevated temperatures e.g. above 40 0 C.
  • a starting aqueous vaccine and the liquid temperature protective agent may be mixed such that the liquid temperature protective agent forms from 1-80% by volume of the final mixture.
  • Suitable temperature protective agents should be safe for human administration, readily miscible/soluble in water, and should not damage other components (e.g. antigen and adjuvant) in the composition.
  • examples include glycerin, propylene glycol, and/or polyethylene glycol (PEG).
  • PEGs may have an average molecular weight ranging from 200-20,000 Da.
  • the polyethylene glycol can have an average molecular weight of about 300 Da ('PEG-300').
  • the invention provides an immunogenic composition comprising: (i) one or more antigen(s) selected from the first, second, third or fourth antigen groups; and (ii) a temperature protective agent.
  • This composition may be formed by mixing (i) an aqueous composition comprising one or more antigen(s) selected from the first, second, third or fourth antigen groups, with (ii) a temperature protective agent. The mixture may then be stored e.g. below 0 0 C, from 0-20°C, from 20-35 °C, from
  • composition may alternatively be formed by mixing (i) a dried composition comprising one or more antigen(s) selected from the first, second, third or fourth antigen groups, with (ii) a liquid composition comprising the temperature protective agent.
  • component (ii) can be used to reconstitute component (i).
  • the invention also provides a method for raising an immune response in a mammal comprising the step of administering an effective amount of a composition of the invention.
  • the immune response is preferably protective and preferably involves antibodies and/or cell-mediated immunity.
  • the method may raise a booster response.
  • the invention also provides at least two antigens of the invention for combined use as a medicament e.g. for use in raising an immune response in a mammal.
  • the invention also provides the use of at least two antigens of the invention in the manufacture of a medicament for raising an immune response in a mammal.
  • the mammal By raising an immune response in the mammal by these uses and methods, the mammal can be protected against S.aureus infection, including a nosocomial infection. More particularly, the mammal may be protected against a skin infection, pneumonia, meningitis, osteomyelitis endocarditis, toxic shock syndrome, and/or septicaemia.
  • the invention also provides a kit comprising a first component and a second component wherein neither the first component nor the second component is a composition of the invention as described above, but wherein the first component and the second component can be combined to provide a composition of the invention as described above.
  • the kit may further include a third component comprising one or more of the following: instructions, syringe or other delivery device, adjuvant, or pharmaceutically acceptable formulating solution.
  • the invention also provides a delivery device pre-filled with an immunogenic composition of the invention.
  • the mammal is preferably a human.
  • the human is preferably a child (e.g. a toddler or infant) or a teenager; where the vaccine is for therapeutic use, the human is preferably a teenager or an adult.
  • a vaccine intended for children may also be administered to adults e.g. to assess safety, dosage, immunogenicity, etc.
  • Other mammals which can usefully be immunised according to the invention are cows, dogs, horses, and pigs.
  • One way of checking efficacy of therapeutic treatment involves monitoring S.aureus infection after administration of the compositions of the invention.
  • One way of checking efficacy of prophylactic treatment involves monitoring immune responses, systemically (such as monitoring the level of IgGl and IgG2a production) and/or mucosally (such as monitoring the level of IgA production), against the antigens in the compositions of the invention after administration of the composition.
  • antigen-specific serum antibody responses are determined post-immunisation but pre-challenge whereas antigen-specific mucosal antibody responses are determined post-immunisation and post- challenge.
  • Another way of assessing the immunogenicity of the compositions of the present invention is to express the proteins recombinantly for screening patient sera or mucosal secretions by immunoblot and/or microarrays. A positive reaction between the protein and the patient sample indicates that the patient has mounted an immune response to the protein in question. This method may also be used to identify immunodominant antigens and/or epitopes within antigens.
  • the efficacy of vaccine compositions can also be determined in vivo by challenging animal models of S. aureus infection, e.g., guinea pigs or mice, with the vaccine compositions, hi particular, there are three useful animal models for the study of S. aureus infectious disease, namely: (i) the murine abscess model [207], (ii) the murine lethal infection model [207] and (iii) the murine pneumonia model [208].
  • the abscess model looks at abscesses in mouse kidneys after intravenous challenge.
  • the lethal infection model looks at the number of mice which survive after being infected by a normally-lethal dose of S. aureus by the intravenous or intraperitoneal route.
  • compositions of the invention will generally be administered directly to a patient. Direct delivery may be accomplished by parenteral injection ⁇ e.g. subcutaneously, intraperitoneally, intravenously, intramuscularly, or to the interstitial space of a tissue), or mucosally, such as by rectal, oral (e.g.
  • the invention may be used to elicit systemic and/or mucosal immunity, preferably to elicit an enhanced systemic and/or mucosal immunity.
  • the enhanced systemic and/or mucosal immunity is reflected in an enhanced THl and/or TH2 immune response.
  • the enhanced immune response includes an increase in the production of IgGl and/or IgG2a and/or IgA.
  • Dosage can be by a single dose schedule or a multiple dose schedule. Multiple doses may be used in a primary immunisation schedule and/or in a booster immunisation schedule, hi a multiple dose schedule the various doses may be given by the same or different routes e.g. a parenteral prime and mucosal boost, a mucosal prime and parenteral boost, etc. Multiple doses will typically be administered at least 1 week apart ⁇ e.g.
  • Vaccines prepared according to the invention may be used to treat both children and adults.
  • a human patient may be less than 1 year old, 1-5 years old, 5-15 years old, 15-55 years old, or at least 55 years old.
  • Preferred patients for receiving the vaccines are the elderly (e.g. >50 years old, >60 years old, and preferably >65 years), the young (e.g. ⁇ 5 years old), hospitalised patients, healthcare workers, armed service and military personnel, pregnant women, the chronically ill, or immunodef ⁇ cient patients.
  • the vaccines are not suitable solely for these groups, however, and may be used more generally in a population.
  • Vaccines produced by the invention may be administered to patients at substantially the same time as (e.g. during the same medical consultation or visit to a healthcare professional or vaccination centre) other vaccines e.g. at substantially the same time as an influenza vaccine, a measles vaccine, a mumps vaccine, a rubella vaccine, a MMR vaccine, a varicella vaccine, a MMRV vaccine, a diphtheria vaccine, a tetanus vaccine, a pertussis vaccine, a DTP vaccine, a conjugated H.influenzae type b vaccine, an inactivated poliovirus vaccine, a hepatitis B virus vaccine, a meningococcal conjugate vaccine (such as a tetravalent A-C-Wl 35-Y vaccine), a respiratory syncytial virus vaccine, etc.
  • Further non-staphylococcal vaccines suitable for co-administration may include one or more antigens listed on pages 33-46 of reference 51.
  • the immunogenic compositions described above include polypeptide antigens from S. aureus. In all cases, however, the polypeptide antigens can be replaced by nucleic acids (typically DNA) encoding those polypeptides, to give compositions, methods and uses based on nucleic acid immunisation. Nucleic acid immunisation is now a developed field (e.g. see references 209 to 216 etc.).
  • the nucleic acid encoding the immunogen is expressed in vivo after delivery to a patient and the expressed immunogen then stimulates the immune system.
  • the active ingredient will typically take the form of a nucleic acid vector comprising: (i) a promoter; (ii) a sequence encoding the immunogen, operably linked to the promoter; and optionally (iii) a selectable marker.
  • Preferred vectors may further comprise (iv) an origin of replication; and (v) a transcription terminator downstream of and operably linked to (ii).
  • (i) & (v) will be eukaryotic and (iii) & (iv) will be prokaryotic.
  • Preferred promoters are viral promoters e.g. from cytomegalovirus (CMV).
  • the vector may also include transcriptional regulatory sequences (e.g. enhancers) in addition to the promoter and which interact functionally with the promoter.
  • Preferred vectors include the immediate-early CMV enhancer/promoter, and more preferred vectors also include CMV intron A.
  • the promoter is operably linked to a downstream sequence encoding an immunogen, such that expression of the immunogen-encoding sequence is under the promoter's control.
  • a marker preferably functions in a microbial host (e.g. in a prokaryote, in a bacteria, in a yeast).
  • the marker is preferably a prokaryotic selectable marker (e.g. transcribed under the control of a prokaryotic promoter).
  • typical markers are antibiotic resistance genes.
  • the vector of the invention is preferably an autonomously replicating episomal or extrachromosomal vector, such as a plasmid.
  • the vector of the invention preferably comprises an origin of replication. It is preferred that the origin of replication is active in prokaryotes but not in eukaryotes.
  • Preferred vectors thus include a prokaryotic marker for selection of the vector, a prokaryotic origin of replication, but a eukaryotic promoter for driving transcription of the immunogen-encoding sequence.
  • the vectors will therefore (a) be amplified and selected in prokaryotic hosts without polypeptide expression, but (b) be expressed in eukaryotic hosts without being amplified. This arrangement is ideal for nucleic acid immunization vectors.
  • the vector of the invention may comprise a eukaryotic transcriptional terminator sequence downstream of the coding sequence. This can enhance transcription levels.
  • the vector of the invention preferably comprises a polyadenylation sequence.
  • a preferred polyadenylation sequence is from bovine growth hormone.
  • the vector of the invention may comprise a multiple cloning site
  • the vector may comprise a second eukaryotic coding sequence.
  • the vector may also comprise an IRES upstream of said second sequence in order to permit translation of a second eukaryotic polypeptide from the same transcript as the immunogen.
  • the immunogen-coding sequence may be downstream of an IRES.
  • the vector of the invention may comprise unmethylated CpG motifs e.g. unmethylated DNA sequences which have in common a cytosine preceding a guanosine, flanked by two 5' purines and two 3' pyrimidines. In their unmethylated form these DNA motifs have been demonstrated to be potent stimulators of several types of immune cell.
  • Vectors may be delivered in a targeted way.
  • Receptor-mediated DNA delivery techniques are described in, for example, references 217 to 222.
  • Therapeutic compositions containing a nucleic acid are administered in a range of about lOOng to about 200mg of DNA for local administration in a gene therapy protocol. Concentration ranges of about 500 ng to about 50 mg, about l ⁇ g to about 2 mg, about 5 ⁇ g to about 500 ⁇ g, and about 20 ⁇ g to about lOO ⁇ g of DNA can also be used during a gene therapy protocol.
  • Factors such as method of action (e.g. for enhancing or inhibiting levels of the encoded gene product) and efficacy of transformation and expression are considerations which will affect the dosage required for ultimate efficacy.
  • Vectors can be delivered using gene delivery vehicles.
  • the gene delivery vehicle can be of viral or non-viral origin (see generally references 223 to 226).
  • Viral-based vectors for delivery of a desired nucleic acid and expression in a desired cell are well known in the art.
  • Exemplary viral-based vehicles include, but are not limited to, recombinant retroviruses (e.g. references 227 to 237), alphavirus-based vectors (e.g.
  • Sindbis virus vectors Semliki forest virus (ATCC VR-67; ATCC VR-1247), Ross River virus (ATCC VR-373; ATCC VR-1246) and Venezuelan equine encephalitis virus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-532); hybrids or chimeras of these viruses may also be used), poxvirus vectors (e.g. vaccinia, fowlpox, canarypox, modified vaccinia Ankara, etc.), adenovirus vectors, and adeno- associated virus (AAV) vectors (e.g. see refs. 238 to 243).
  • AAV adeno- associated virus
  • Non-viral delivery vehicles and methods can also be employed, including, but not limited to, polycationic condensed DNA linked or unlinked to killed adenovirus alone [e.g. 244], ligand-linked
  • DNA [245] eukaryotic cell delivery vehicles cells [e.g. refs. 246 to 250] and nucleic charge neutralization or fusion with cell membranes. Naked DNA can also be employed. Exemplary naked
  • Liposomes e.g. immunoliposomes
  • Additional approaches are described in references 258 & 259.
  • non- viral delivery suitable for use includes mechanical delivery systems such as the approach described in ref. 259.
  • the coding sequence and the product of expression of such can be delivered through deposition of photopolymerized hydrogel materials or use of ionizing radiation [e.g. refs. 260 & 261].
  • Other conventional methods for gene delivery that can be used for delivery of the coding sequence include, for example, use of hand-held gene transfer particle gun [262] or use of ionizing radiation for activating transferred genes [260 & 261].
  • PLG poly(lactide-co-glycolide) ⁇ microparticles
  • Delivery DNA using PLG is a particularly preferred method e.g. by adsorption to the microparticles, which are optionally treated to have a negatively- charged surface (e.g. treated with SDS) or a positively-charged surface (e.g. treated with a cationic detergent, such as CTAB).
  • a negatively- charged surface e.g. treated with SDS
  • CTAB a positively-charged surface
  • SEQ ID NO: 234 is the 'iron (Fe+3) ABC superfamily ATP binding cassette transporter, binding protein' from S.epidermidis strain M23864:W1, with 73% identity to SEQ ID NO: 42 (staOO ⁇ ), and SEQ ID NO: 235 is the 'putative lipoprotein' from S.epidermidis strain RP62A, with 67% identity to SEQ ID NO: 47 (staOl 1).
  • S.epidermidis is commonly present on human skin and can sometimes cause illness.
  • the invention provides an immunogenic composition comprising:
  • polypeptide comprising an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 234 comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 234, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more); and/or
  • polypeptide comprising an amino acid sequence: (a) having 50% or more identity (e.g.
  • SEQ ID NO: 235 SEQ ID NO: 235; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 235, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50,
  • composition may also include an adjuvant. These compositions are particularly useful for immunising a mammal (including a human) against S.epidermidis infection.
  • Preferred fragments of (b) comprise an epitope from SEQ ID NO: 234 or 235, respectively.
  • Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 234/235 while retaining at least one epitope of SEQ ID NO: 234/235.
  • the invention provides the use of the sta006 and/or staOl 1 homolog from any Staphylococcus species for immunising a mammal against that species.
  • Antibodies against S.aureus antigens can be used for passive immunisation.
  • the invention provides an antibody which is specific for an antigen in the first, second, third or fourth antigen groups.
  • the invention also provides the use of such antibodies in therapy.
  • the invention also provides the use of such antibodies in the manufacture of a medicament.
  • the invention also provides a method for treating a mammal comprising the step of administering an effective amount of an antibody of the invention. As described above for immunogenic compositions, these methods and uses allow a mammal to be protected against S.aureus infection.
  • the term "antibody" includes intact immunoglobulin molecules, as well as fragments thereof which are capable of binding an antigen.
  • antibodies include hybrid (chimeric) antibody molecules [263, 264]; F(ab')2 and F(ab) fragments and Fv molecules; non-covalent heterodimers [265, 266]; single-chain Fv molecules (sFv) [267]; dimeric and trimeric antibody fragment constructs; minibodies [268, 269]; humanized antibody molecules [270-272]; and any functional fragments obtained from such molecules, as well as antibodies obtained through non-conventional processes such as phage display.
  • the antibodies are monoclonal antibodies. Methods of obtaining monoclonal antibodies are well known in the art. Humanised or fully-human antibodies are preferred.
  • GI numbering is used above.
  • a GI number, or “Genlnfo Identifier” is a series of digits assigned consecutively to each sequence record processed by NCBI when sequences are added to its databases. The GI number bears no resemblance to the accession number of the sequence record.
  • a sequence is updated (e.g. for correction, or to add more annotation or information) then it receives a new GI number. Thus the sequence associated with a given GI number is never changed.
  • this epitope may be a B-cell epitope and/or a T-cell epitope.
  • Such epitopes can be identified empirically (e.g. using PEPSCAN [281,282] or similar methods), or they can be predicted (e.g. using the Jameson- Wolf antigenic index [283], matrix-based approaches [284], MAPITOPE [285], TEPITOPE [286,287], neural networks [288], OptiMer & EpiMer [289, 290], ADEPT [291], Tsites [292], hydrophilicity [293], antigenic index [294] or the methods disclosed in references 295-299, etc.).
  • Epitopes are the parts of an antigen that are recognised by and bind to the antigen binding sites of antibodies or T-cell receptors, and they may also be referred to as "antigenic determinants”.
  • an antigen "domain” is omitted, this may involve omission of a signal peptide, of a cytoplasmic domain, of a transmembrane domain, of an extracellular domain, etc.
  • composition comprising
  • X may consist exclusively of X or may include something additional e.g. X + Y.
  • the term "about” in relation to a numerical value JC is optional and means, for example, jc+10%.
  • references to a percentage sequence identity between two amino acid sequences means that, when aligned, that percentage of amino acids are the same in comparing the two sequences.
  • This alignment and the percent homology or sequence identity can be determined using software programs known in the art, for example those described in section 7.7.18 of ref. 300.
  • a preferred alignment is determined by the Smith- Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62.
  • the Smith- Waterman homology search algorithm is disclosed in ref. 301.
  • Figure 1 shows bacterial counts (Log cfu/ml) after challenge of mice previously immunised with the indicated antigens.
  • Figures 2 to 4 show survival (%) after challenge of mice previously immunised with various mixtures of antigens over 14 days.
  • the six groups from SA-10-a are, from top to bottom at day 14.
  • groups (i), (iii) & (iv) together, (ii), IsdB, then the negative control, hi Figure 3 the six groups from SA-10-a are, from top to bottom at day 14.
  • the six groups from SA-10-b are, from top to bottom at day 14.
  • Figure 5 shows collected data on mouse survival from four different experiments after challenge of mice previously immunised with various compositions (PBS negative control; IsdB antigen; and "Combo- 1" and "Combo-2” antigen combinations of the invention). Individual symbols show the survival duration of individual mice; the horizontal bar for each group shows the median survival duration; the percentage figures are survival 14 days after challenge; and the p values at the top are t-Test comparisons of median survival durations between groups.
  • Figure 6 shows the number of colony forming units (cfu) in mouse kidneys after infection with 9x10 6 cfu of Newman strain in the abscess model. Horizontal bars are averages per group, and the figure beneath each group is the log reduction relative to the PBS control group.
  • Figure 7 shows bacterial count (log CFU/ml) in kidneys of mice in an abscess model experiment. Mice were challenged with the following strains: (A) MW2; (B) LAC; (C) Staphl9; or (D) MU50. Each point is an individual animal and the bar shows the median count per group. Mice had been immunised as shown on the x-axis label.
  • Figure 8 shows the formation of StaOl l oligomers in the presence of increasing concentrations of Ca + * ions. Numbers indicate mM concentrations, and a * indicates the presence of 5OmM EDTA.
  • Figure 9 shows IgG titers against (A) EsxAB (B) Sta006 (C) Hla-H35L (D) StaOl 1.
  • Each graph has three groups, with a pair of bars per group. The right-hand bar in a pair shows pre-immune IgG and the left-hand bar shows post-immune IgG.
  • the three groups are the compositions used for immunising and, from left to right, are: negative control of adjuvant alone; the Combo 1 combination; and the relevant antigen alone.
  • Figure 10 shows bacterial counts values (log CFU/ml) in mice after challenge with the indicated strains. Each point is an individual animal and the bar shows the median CFU. The P value beneath the IsdB and Combo columns is a comparison against the adjuvant-only control.
  • Figure 11 shows the area of abscesses (mm 2 ) in mice after challenge with Newman strain.
  • Figure 12 shows days of survival of mice after challenge with four different strains: Newman (o), ST-80 (D), USA300-FPR3757 ( ⁇ ) or USA300-Lac (x) strains. Each point is an individual animal, the bar shows the median survival, and the heading number shows the % of animals surviving after 15 days. Mice received aluminium hydroxide adjuvant alone, IsdB or Combo 1.
  • Figure 13 shows the median survival (days) of mice after challenge. The mice had been immunised with the antigens indicated on the X-axis. Each point is an individual animal and the bar shows the median survival. The heading numbers show the % of animals surviving after 15 days.
  • S.aureus proteins have been selected for use as vaccine components based on various criteria.
  • IsdA is a surface protein involved in iron uptake. It is detectable with a high molecular weight (>250kDa) in immunoblots of whole cell lysates and cell wall fractions of S. aureus. Furthermore, labelled anti-IsdA antibodies revealed extracellular structures. These structures were seen in a variety of growth and infection conditions, including iron positive conditions (in which IsdA expression is reported to be suppressed). The structures have a tail up to 4 ⁇ m long, with a typical orientation parallel to the mammalian cell surface. Detached IsdA-positive structures were observed to adhere on the surface of epithelial cells, but lose cell junction localization. Epithelia/bacteria interaction may stimulate expression of the structures.
  • IsdA is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. Iron uptake is important for virulence, so the protein is likely to be available for immune attack at pathological stages of the bacterial life cycle.
  • the inventors have found that the protein is not cytotoxic to human cells (see below).
  • the protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
  • EsxA and EsxB are small acidic dimeric secreted proteins.
  • the inventors have found that EsxA is highly conserved between different strains (present in 36/36 strains tested; see below), while EsxB is present in 25/36 strains.
  • the proteins are involved in persisting an infection and so are likely to be available for immune attack at pathological stages of the bacterial life cycle.
  • the inventors have found that a fusion of EsxA and EsxB ('EsxAB') is not cytotoxic to human cells (see below). It can also adsorb well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans.
  • the antigens are useful for providing an immune response to prevent hematic spread of the bacterium.
  • HIa is a pore-forming secreted toxin. This protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. It is an important virulence factor so is likely to be available for immune attack at pathological stages of the bacterial life cycle. It is not cytotoxic to human cells (see below). The protein can adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent pneumonia.
  • Spa is a surface protein involved in Fc binding.
  • the inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested), thus offering protection across a broad population of circulating strains. It is important for virulence so is likely to be available for immune attack at pathological stages of the bacterial life cycle.
  • the protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
  • Sta006 also known as FhuD2
  • FhuD2 is a surface protein involved in iron uptake.
  • the inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains.
  • the inventors have found that the protein is not cytotoxic to human cells (see below).
  • the protein can also adsorb well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
  • StaOl 1 is a surface lipoprotein.
  • the inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains.
  • the inventors have found that the protein is not cytotoxic to human cells (see below).
  • the protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
  • This protein has been shown to assemble into oligomers in the presence of Ca +4 ions, but not Mg "1"1" ions (see Figure 8).
  • Conjugated capsular saccharides are useful for providing opsonic immunity.
  • Serotypes 5 and 8 cover about 85% of clinical isolates.
  • a panel of 36 clinical isolates was used to represent circulating strains, including strains belonging to the five clonal lineages representing the vast majority of worldwide circulating CA-MRSA (community-associated methicillin-resistant S.aureus).
  • HA-MRSA hospital-associated MRSA
  • non-MRSA strains were also included.
  • Overall the panel included 9 HA-MRSA strains, 7 CA-MRSA strains, 2 MRSA strains, and 18 other strains.

Abstract

An effective Staphylococcus aureus vaccine may require several antigenic components, and so various combinations of S. aureus antigens are identified for use in immunisation. These polypeptides may optionally be used in combination with S.aureus saccharides.

Description

COMPOSITIONS FOR IMMUNISING AGAINST STAPHYLOCOCCUS AUREUS
TECHNICAL FIELD
This invention relates to antigens derived from S.aureus and to their use in immunisation.
BACKGROUND ART Staphylococcus aureus is a Gram-positive spherical bacterium. Annual US mortality exceeds that of any other infectious disease, including HIV/ AIDS, and S.aureus is the leading cause of bloodstream, lower respiratory tract, skin & soft tissue infections. There is currently no authorised vaccine. A vaccine based on a mixture of surface polysaccharides from bacterial types 5 and 8, StaphVAX™, failed to reduce infections when compared to the placebo group in a phase III clinical trial in 2005. Reference 1 reports that the "V710" vaccine from Merck and Intercell is undergoing a phase 2/3 trial on patients undergoing cardiothoracic surgery. The V710 vaccine is based on a single antigen, IsdB [2], a conserved iron-sequestering cell-surface protein.
S.aureus causes a range of illnesses from minor skin infections to life-threatening diseases such as pneumonia, meningitis, osteomyelitis, bacteremia, endocarditis, toxic shock syndrome, organ abscesses and septicemia. The bacterium has multiple virulence factors which are differentially expressed during different phases of its life cycle, and so a vaccine which can prevent one disease might not prevent another. For instance, the V710 vaccine may be effective against hematic spread of the S.aureus, but may be ineffective against pneumonia and may not elicit any opsonic activity. One aim of the invention is to provide vaccines which can protect against hematic spread and pneumonia, and which may also elicit an opsonic response.
Thus there remains a need to identify further and improved antigens for use in S.aureus vaccines, and in particular for vaccines which are useful against multiple S.aureus pathologies.
DISCLOSURE OF THE INVENTION
The inventors have identified various S.aureus polypeptides that are useful for immunisation, either alone or in combination. These polypeptides may be combined with S.aureus saccharides or other S.aureus polypeptides. The antigens are useful in S.aureus vaccines but may also be used as components in vaccines for immunising against multiple pathogens.
The inventors have identified the following 36 polypeptides: clfA, clfB, coA, eap, ebhA, ebpS, efb, emp, esaC, esxA, esxB, FnBA, FnBB, HIa, hlgB, hlgC, isdA, isdB, isdC, isdG, isdH, isdl, lukD, lukE, lukF, lukS, nuc, sasA, sasB, sasC, sasD, sasF, sdrC, sdrD, spa, and sdrE2. This set of antigens is referred to herein as 'the first antigen group'. Thus the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a clfA antigen; (2) a clfB antigen; (3) a coA antigen; (4) a eap antigen; (5) a ebhA antigen; (6) a ebpS antigen; (7) a efb antigen; (8) a emp antigen; (9) a esaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) a FnBA antigen; (13) a FnBB antigen; (14) a HIa antigen; (15) a hlgB antigen; (16) a hlgC antigen; (17) a isdA antigen; (18) a isdB antigen; (19) a isdC antigen; (20) a isdG antigen; (21) a isdH antigen; (22) a isdl antigen; (23) a lukD antigen; (24) a lukE antigen; (25) a lukF antigen; (26) a lukS antigen; (27) a nuc antigen; (28) a sasA antigen; (29) a sasB antigen; (30) a sasC antigen; (31) a sasD antigen; (32) a sasF antigen; (33) a sdrC antigen; (34) a sdrD antigen; (35) a spa antigen; (36) a sdrE2 antigen.
Within the first antigen group, antigens are preferably selected from a subset of 16 of the 36 polypeptides, namely: clfA, clfB, emp, esaC, esxA, esxB, hla, isdA, isdB, isdC, sasD, sasF, sdrC, sdrD, spa, and sdrE2. Thus the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of these sixteen antigens.
The inventors have also identified the following 128 polypeptides: staOOl, sta002, staOO3, staOO4, staOO5, sta006, sta007, staOO8, sta009, staOlO, staOl l, staO12, staO13, staO14, staO15, staOlό, staO17, staO18, staO19, sta020, staO21, staO22, staO23, staO24, staO25, staO26, staO27, staO28, staO29, staO3O, staO31, staO32, staO33, staO34, staO35, staO36, staO37, staO38, staO39, staO4O, staO41, staO42, staO43, staO44, staO45, staO46, staO47, staO48, staO49, staO5O, staO51, staO52, staO53, staO54, staO55, staO56, staO57, staO58, staO59, sta060, staOόl, staO62, staO63, staO64, staO65, staO66, staO67, staO68, staO69, sta070, staO71, staO72, staO73, staO74, staO75, staO76, staO77, staO78, staO79, staO8O, staO81, staO82, staO83, staO84, staO85, staO86, staO87, staO88, staO89, sta090, staO91, staO92, staO93, staO94, staO95, staO96, staO97, staO98, staO99, stalOO, stalOl, stalO2, stalO3, stalO4, stalO5, stalO6, stalO7, stalO8, stalO9, stal lO, stal l l, stal l2, stal l3, stal l4, stall5, stal lό, stal l7, stal l8, stal l9, stal20, NW 6, NW 9, NW_10, NWJ, NW_8, NWJ, NW l, and NWJ. This set of antigens is referred to herein as 'the second antigen group'. Thus the invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a staOOl antigen; (2) a sta002 antigen; (3) a staOO3 antigen; (4) a sta004 antigen; (5) a staOO5 antigen; (6) a sta006 antigen; (7) a sta007 antigen; (8) a staOO8 antigen; (9) a sta009 antigen; (10) a staOlO antigen; (11) a staOl l antigen; (12) a staO12 antigen; (13) a staO13 antigen; (14) a staO14 antigen; (15) a staO15 antigen; (16) a staOlό antigen; (17) a staO17 antigen; (18) a staO18 antigen; (19) a staO19 antigen; (20) a sta020 antigen; (21) a staO21 antigen; (22) a staO22 antigen; (23) a staO23 antigen; (24) a staO24 antigen; (25) a staO25 antigen; (26) a staO26 antigen; (27) a staO27 antigen; (28) a staO28 antigen; (29) a staO29 antigen; (30) a staO3O antigen; (31) a staO31 antigen; (32) a staO32 antigen; (33) a staO33 antigen; (34) a staO34 antigen; (35) a staO35 antigen; (36) a staO36 antigen; (37) a staO37 antigen; (38) a staO38 antigen; (39) a staO39 antigen; (40) a sta040 antigen; (41) a staO41 antigen; (42) a staO42 antigen; (43) a staO43 antigen; (44) a staO44 antigen; (45) a staO45 antigen; (46) a staO46 antigen; (47) a staO47 antigen; (48) a staO48 antigen; (49) a staO49 antigen; (50) a staO5O antigen; (51) a staO51 antigen; (52) a staO52 antigen; (53) a staO53 antigen; (54) a staO54 antigen; (55) a staO55 antigen;
(56) a staO56 antigen; (57) a staO57 antigen; (58) a staO58 antigen; (59) a staO59 antigen; (60) a sta060 antigen; (61) a staOόl antigen; (62) a staO62 antigen; (63) a staO63 antigen; (64) a staO64 antigen; (65) a staO65 antigen; (66) a staO66 antigen; (67) a staO67 antigen; (68) a staO68 antigen; (69) a staO69 antigen; (70) a sta070 antigen; (71) a staO71 antigen; (72) a staO72 antigen; (73) a staO73 antigen; (74) a staO74 antigen; (75) a staO75 antigen; (76) a staO76 antigen; (77) a staO77 antigen; (78) a staO78 antigen; (79) a staO79 antigen; (80) a staO8O antigen; (81) a staO81 antigen; (82) a staO82 antigen; (83) a staO83 antigen; (84) a staO84 antigen; (85) a staO85 antigen; (86) a staO86 antigen; (87) a staO87 antigen; (88) a staO88 antigen; (89) a staO89 antigen; (90) a sta090 antigen; (91) a staO91 antigen; (92) a staO92 antigen; (93) a staO93 antigen; (94) a staO94 antigen; (95) a staO95 antigen; (96) a staO96 antigen; (97) a staO97 antigen; (98) a staO98 antigen; (99) a staO99 antigen; (100) a stalOO antigen; (101) a stalOl antigen; (102) a stalO2 antigen; (103) a stalO3 antigen; (104) a stalO4 antigen; (105) a stalO5 antigen; (106) a stalO6 antigen; (107) a stalO7 antigen; (108) a stalO8 antigen; (109) a stalO9 antigen; (110) a stal lO antigen; (111) a stal l l antigen; (112) a stal l2 antigen; (113) a stal l3 antigen; (114) a stal l4 antigen; (115) a stal l5 antigen; (116) a stal lό antigen; (117) a stal l7 antigen; (118) a stal lδ antigen; (119) a stal l9 antigen; (120) a stal20 antigen; (121) a NW 6 antigen; (122) a NW 9 antigen; (123) a NW lO antigen; (124) a NWJ antigen; (125) a NWJ antigen; (126) a NWJ antigen; (127) a NW l antigen; and (128) a NW_5 antigen.
Within the second antigen group of 128 antigens, a preferred subset of 113 antigens omits (81) and (107) to (120) from this list. Within the second antigen group, a subset of 27 of the 128 polypeptides is referred to herein as 'the third antigen group', namely: staOOl, sta002, sta003, sta004, staOO5, sta006, sta007, staOO8, sta009, staOlO, staO19, staO28, sta040, staO49, staO57, staO64, staO73, staO95, staO98, stalOl, stalO5, NW l, NW 6, NWJ, NWJ, NW 9 and NW lO. The invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more {i.e. 2, 3, 4, 5, 6 or more) antigens selected from the third antigen group.
The 101 antigens that are in the second antigen group but not in the third antigen group are referred to herein as 'the fourth antigen group'. Within the fourth antigen group of 101 antigens, a preferred subset of 86 antigens omits (81) and (107) to (120) from the above list. The second antigen group thus consists of a combination of the third and fourth antigen groups. Within the second antigen group, a subset of 8 of the 128 polypeptides is referred to herein as 'the fifth antigen group', namely: staOO4, sta006, sta007, staOl l, staO28, sta060, staO98 and stal l2. The invention provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more {i.e. 2, 3, 4, 5, 6 or more) antigens selected from the fifth antigen group. Within the 36 antigens of the first antigen group there are 630 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 630 pairs.
Within the 128 antigens of the second antigen group there are 8128 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 8128 pairs.
Within the preferred 113 antigens of the second antigen group there are 6328 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 6328 pairs.
Within the preferred 27 antigens of the third antigen group there are 351 possible pairs of different antigens. AU such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 351 pairs. Within the 101 antigens of the fourth antigen group there are 5050 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 5050 pairs. Within the preferred 86 antigens of the fourth antigen group there are 3655 possible pairs of different antigens. All such pairs are disclosed herein and are part of the invention. Thus the invention provides an immunogenic composition comprising a pair of antigens, wherein said pair is one of said 3655 pairs.
In one embodiment, a composition includes at least one antigen (i.e. 1, 2, 3, 4, 5, 6 or more) selected from the first antigen group and at least one antigen (i.e. 1, 2, 3, 4, 5, 6 or more) selected from the second antigen group. Antigens from the first antigen group may be selected from the preferred subset of 16 antigens, and antigens from the second antigen group may be selected from the third antigen group or the fifth antigen group.
The invention also provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigens selected from the group consisting of: (1) a clfA antigen; (2) a clfB antigen; (3) a sdrE2 antigen; (4) a sdrC antigen; (5) a
SasF antigen; (6) a emp antigen; (7) a sdrD antigen; (8) a spa antigen; (9) a esaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) a sta006 antigen; (13) a isdC antigen; (14) a hla antigen; (15) a staOl 1 antigen; (16) isdA antigen; (17) a isdB antigen; (18) a sasF antigen. This group of 18 antigens is sometimes referred to herein as the 'sixth antigen group'. The invention also provides an immunogenic composition comprising a combination of antigens, said combination comprising two or more (i.e. 2, 3, 4 or 5) antigens selected from the group consisting of: (1) a esxA antigen; (2) a esxB antigen; (3) a staOOό antigen; (4) a hla antigen; and/or (5) a staOl 1 antigen. The composition may also include an adjuvant e.g. an aluminium hydroxide adjuvant.
Advantageous combinations of the invention are those in which two or more antigens act synergistically. Thus the protection against S. aureus disease achieved by their combined administration exceeds that expected by mere addition of their individual protective efficacy.
Specific combinations of interest include, but are not limited to: (1) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen and a isdC antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an SdrDE hybrid with an sdrE2 antigen downstream of a sdrD antigen.
(2) An immunogenic composition comprising a sasD antigen, a clfB antigen and a sdrC antigen.
(3) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a clfA antigen.
(4) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a staOl 1 antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
(5) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a staOOό antigen.
(6) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a hla antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen. The Hla antigen may be a detoxified mutant e.g. including a H35L mutation. (7) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a esxA antigen.
(8) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen and a hla antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
(9) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a esxA antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen. (10) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen and a staOl l antigen. The esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
(11) An immunogenic composition comprising a esxA antigen, a esxB antigen and a staOl l antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
(12) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a spa antigen.
(13) An immunogenic composition comprising a esxA antigen, a esxB antigen, a isdA antigen, a staOOό antigen, a staOl 1 antigen and a spa antigen. The esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. The spa antigen may be a fragment of a full- length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
(14) An immunogenic composition comprising a esxA antigen, a esxB antigen, a HIa antigen, a staOOό antigen and a staOl l antigen. The esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
(15) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a sdrE2 antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.
(16) An immunogenic composition comprising a esxA antigen, a esxB antigen and a hla antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation. (17) An immunogenic composition comprising a hla antigen, a isdA antigen, a staOOό antigen and a staOl 1 antigen. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. The Hla antigen may be a detoxified mutant e.g. including a H35L mutation.
(18) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen and a isdA antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
(19) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen and a hla antigen. The Hla antigen may be a detoxified mutant e.g. including a H35L mutation. (20) An immunogenic composition comprising a HIa antigen, a staOOό antigen and a staOl l antigen. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
(21) An immunogenic composition comprising a esxA antigen and a esxB antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid with an esxB antigen downstream of an esxA antigen.
(22) An immunogenic composition comprising a esxA antigen, a esxB antigen and a staOOό antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
(23) An immunogenic composition comprising a spa antigen, a staOOό antigen and a staOl l antigen. The spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
(24) An immunogenic composition comprising a esxA antigen, a esxB antigen, a isdA antigen, a staOOό antigen and a staOl l antigen. The esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
(25) An immunogenic composition comprising a staOOό antigen and a staOl 1 antigen.
(26) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen, a isdA antigen and a clfB antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. The clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163.
(27) An immunogenic composition comprising a staOOό antigen, a staOl l antigen and a staO19 antigen.
(28) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen, a hla antigen and a clfB antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
(29) An immunogenic composition comprising a staOOό antigen, a staOl l antigen, a staO19 antigen, and a hla antigen. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation.
(30) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen, a staOl 1 antigen and a clfB antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The clfB antigen may be a fragment of a full-length clfB antigen e.g. SEQ ID NO: 163.
(31) An immunogenic composition comprising a spa antigen, a esxA antigen, a esxB antigen, a staOOό antigen and a staOl 1 antigen. The spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc. The esxA and esxB antigens may be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid.
(32) An immunogenic composition comprising a sdrD antigen, a sdrE2 antigen, a isdC antigen and a esxB antigen. The sdrD and sdrE2 antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen. (33) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen, a staOl l antigen and a staO19 antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.
(34) An immunogenic composition comprising a esxA antigen, a esxB antigen, a staOOό antigen, a isdA antigen and a sdrD antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. The sdrD antigen may be a fragment of a full-length sdrD antigen e.g. SEQ ID NO: 156.
(35) An immunogenic composition comprising a esxA antigen, a esxB antigen, and a isdA antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen. The isdA antigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157.
(36) An immunogenic composition comprising a sasD antigen, a clfB antigen, a sdrC antigen, a esxA antigen and a esxB antigen. The esxA and esxB antigens can usefully be combined as a hybrid polypeptide, as discussed below, e.g. an EsxAB hybrid with an esxB antigen downstream of an esxA antigen.
(37) An immunogenic composition comprising a HIa antigen, a spa antigen, a staOOό antigen and a staOl 1 antigen. The HIa antigen may be a detoxified mutant e.g. including a H35L mutation. The spa antigen may be a fragment of a full-length spa antigen, such as a Spa(D) domain mutated to disrupt or decrease binding to IgG Fc.
In some embodiments, any of these 37 compositions may include additional staphylococcal antigens, and these further antigens can be polypeptides and/or saccharides. For example, they can usefully also include one or more S.αureus capsular saccharide coηjugate(s) e.g. against a serotype 5 and/or a serotype 8 strain. The inclusion of one or both such conjugates is particularly useful for combinations (8), (10), (20), (23), (25), (31) and (37). In other embodiments, these 37 compositions include no additional staphylococcal polypeptide antigens, hi other embodiments, these 37 compositions include no additional staphylococcal antigens, hi other embodiments, these 37 compositions include no additional antigens.
The invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 151; and/or (b) comprising a fragment of at least 'n' consecutive amino acids from amino acids 1-97 of SEQ ID NO: 151 and at least 'n' consecutive amino acids from amino acids 104- 207 of SEQ ID NO: 151, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). The invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 152; and/or (b) comprising a fragment of at least 'n' consecutive amino acids from amino acids 1-104 of SEQ ID NO: 152 and at least 'n' consecutive amino acids from amino acids 111-207 of SEQ ID NO: 152, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These polypeptides can elicit antibodies (e.g. when administered to a human) which recognise both the wild-type staphylococcal protein comprising SEQ ID NO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO: 11. Thus the immune response will recognise both of antigens esxA and esxB. Preferred fragments of (b) provide an epitope from SEQ ID NO: 10 and an epitope from SEQ ID NO: 11. The invention also provides an immunogenic composition comprising a combination of such a protein and an adjuvant, such as an aluminium hydroxide adjuvant.
The invention also provides a polypeptide comprising amino acid sequence (a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 241; and/or (b) comprising both a fragment of at least 'n' consecutive amino acids from amino acids 1-96 of SEQ ID NO: 241 and a fragment of at least 'n' consecutive amino acids from amino acids 103-205 of SEQ ID NO: 241, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These polypeptides (e.g. SEQ ID NO: 250) can elicit antibodies (e.g. when administered to a human) which recognise both the wild-type staphylococcal protein comprising SEQ ID NO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO: 11. Thus the immune response will recognise both of antigens esxA and esxB. Preferred fragments of (b) provide an epitope from SEQ ID NO: 10 and an epitope from SEQ ID NO: 11. The invention also provides an immunogenic composition comprising a combination of such a protein and an adjuvant, such as an aluminium hydroxide adjuvant.
The invention also provides a polypeptide comprising a staphylococcal hemolysin sequence, wherein the sequence does not include a sequence having at least 90% identity to SEQ ID NO: 217 but can elicit antibodies which can kill staphylococci. The polypeptide may have a first sequence having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 218 and a second sequence having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 219, wherein the first and second sequences are either directly joined or are joined by an intervening amino acid sequence having fewer than 40 amino acids (e.g. <35 amino acids, <30 amino acids, <25 amino acids, <20 amino acids, <15 amino acids, <10 amino acids, <5 amino acids). SEQ ID NOs: 189 and 216 are examples of such polypeptides, in which the first and second sequences are joined by a tetrapeptide PSGS sequence (SEQ ID NO: 225).
The invention also provides an immunogenic composition comprising a StaOl l antigen and a Ca+* ion. The antigen and Ca^ ion may form a complex e.g. atoms in the antigen may coordinate the Ca+* ion. The immunogenic composition may also include an adjuvant. The invention also provides a oligomer of a StaOl l antigen, and also immunogenic compositions comprising such oligomers. The oligomer can be a dimer, trimer, tetramer, pentamer, hexamer, heptamer, octamer or higher. An oligomer may comprise a Ca+4^ ion, and a composition comprising StaOl 1 oligomers may comprise 5-50OmM Ca+* ions.
Further polypeptide antigens In additions to antigens from the various antigen groups of the invention, immunogenic compositions may include one or more of the following S. aureus antigens (or antigens comprising immunogenic fragment(s) thereof) to enhance the efficacy against S.aureus of an immune response elicited by the composition [e.g. see references 3-10]:
• AhpC • AhpF
• Autolysin amidase
• Autolysin glucosaminidase
• Collagen binding protein CAN
• EbhB • GehD lipase
• Heparin binding protein HBP (17kDa)
• Laminin receptor
• MAP
• MntC (also known as SitC) • MRPII
• Npase
• ORF0594
• ORF0657n
• ORF0826 • PBP4
• RAP (RNA III activating protein) • Sai-1
• SasK
• SBI
• SdrG • SdrH
• SSP-I
• SSP-2
• Vitronectin-binding protein
Combinations with saccharides The individual antigens identified in the antigen groups of the invention may be used in combination with conjugated saccharide antigens. Thus the invention provides an immunogenic composition comprising a combination of:
(1) one or more antigen(s) selected from the first, second, third or fourth antigen groups (as defined above); and (2) one or more conjugates of a S.aureus exopolysaccharide and a carrier protein.
A conjugate used in component (2) of this combination includes a saccharide moiety and a carrier moiety. The saccharide moiety is from the exopolysaccharide of S.aureus, which is a poly-N- acetylglucosamine (PNAG). The saccharide may be a polysaccharide having the size that arises during purification of the exopolysaccharide from bacteria, or it may be an oligosaccharide achieved by fragmentation of such a polysaccharide e.g. size can vary from over 40OkDa to between 75 and 40OkDa, or between 10 and 75kDa, or up to 30 repeat units. The saccharide moiety can have various degrees of N-acetylation and, as described in reference 11, the PNAG may be less than 40% N-acetylated (e.g. less than 35, 30, 20, 15, 10 or 5% N-acetylated; deacetylated PNAG is also known as dPNAG). Deacetylated epitopes of PNAG can elicit antibodies that are capable of mediating opsonic killing. The PNAG may or may not be O-succinylated e.g. it may be O-succinylated on fewer less than 25, 20, 15, 10, 5, 2, 1 or 0.1% of residues.
The invention also provides an immunogenic composition comprising a combination of:
(1) one or more antigen(s) selected from the first, second, third or fourth antigen groups; and
(2) one or more conjugates of a S.aureus capsular saccharide and a carrier protein. A conjugate used in component (2) of this combination includes a saccharide moiety and a carrier moiety. The saccharide moiety is from the capsular saccharide of a S.aureus. The saccharide may be a polysaccharide having the size that arises during purification of capsular polysaccharide from bacteria, or it may be an oligosaccharide achieved by fragmentation of such a polysaccharide. Capsular saccharides may be obtained from any suitable strain of S.aureus (or any bacterium having a similar or identical saccharide), such as from a type 5 and/or a type 8 S.aureus strain and/or a type
336 S.aureus strain. Most strains of infectious S.aureus contain either Type 5 or Type 8 capsular saccharides. Both have FucNAcp in their repeat unit as well as ManNAcA which can be used to introduce a sulfhydryl group for linkage. The repeating unit of the Type 5 saccharide is →4)-β-D- Man NAcA-(I →4)-α-L-FucNAc(3OAc)-(l — »θ)-β-D-FucNAc-(l→, whereas the repeating unit of the Type 8 saccharide is →3)-β-D-ManNAcA(4OAc)-(l→3)-α-L-FucNAc(l→3)-α-D-FucNAc(l→. The type 336 saccharide is a β-linked hexosamine with no O-acetylation [12,13] and is cross-reactive with antibodies raised against the 336 strain (ATCC 55804). A combination of a type 5 and a type 8 saccharide is typical, and a type 336 saccharide may be added to this pairing [14].
The invention also provides an immunogenic composition comprising a combination of:
(1) one or more antigen(s) selected from the first, second, third or fourth antigen groups; (2) one or more conjugates of a S.aureus exopolysaccharide and a carrier protein; and
(3) one or more conjugates of a S.aureus capsular saccharide and a carrier protein.
The carrier moiety in these conjugates will usually be a protein, but usually not one of the antigens of (1). Typical carrier proteins are bacterial toxins, such as diphtheria or tetanus toxins, or toxoids or mutants or fragments thereof. The CRM 197 diphtheria toxin mutant [15] is useful. Other suitable carrier proteins include the N. meningitidis outer membrane protein complex [16], synthetic peptides [17,18], heat shock proteins [19,20], pertussis proteins [21,22], cytokines [23], lymphokines [23], hormones [23], growth factors [23], artificial proteins comprising multiple human CD4+ T cell epitopes from various pathogen-derived antigens [24] such as N19 [25], protein D from H.influenzae [26-28], pneumolysin [29] or its non-toxic derivatives [30], pneumococcal surface protein PspA [31], iron-uptake proteins [32], toxin A or B from C.difficile [33], recombinant P.aeruginosa exoprotein A (rEPA) [34], etc. In some embodiments the carrier protein is a S.aureus protein, such as an antigen selected from the first, second, third or fourth antigen groups.
Where a composition includes more than one conjugate, each conjugate may use the same carrier protein or a different carrier protein. Conjugates may have excess carrier (w/w) or excess saccharide (w/w). In some embodiments, a conjugate may include substantially equal weights of each.
The carrier molecule may be covalently conjugated to the carrier directly or via a linker. Direct linkages to the protein may be achieved by, for instance, reductive animation between the saccharide and the carrier, as described in, for example, references 35 and 36. The saccharide may first need to be activated e.g. by oxidation. Linkages via a linker group may be made using any known procedure, for example, the procedures described in references 37 and 38. A preferred type of linkage is an adipic acid linker, which may be formed by coupling a free -NH2 group {e.g. introduced to a glucan by amination) with adipic acid (using, for example, diimide activation), and then coupling a protein to the resulting saccharide-adipic acid intermediate [39,40]. Another preferred type of linkage is a carbonyl linker, which may be formed by reaction of a free hydroxyl group of a saccharide CDI [41, 42] followed by reaction with a protein to form a carbamate linkage. Other linkers include β- propionamido [43], nitrophenyl-ethylamine [44], haloacyl halides [45], glycosidic linkages [46], 6- aminocaproic acid [47], ADH [48], C4 to Ci2 moieties [49], etc. Carbodiimide condensation can also be used [50].
PNAG conjugates may be prepared in various ways e.g. by a process comprising: a) activating the PNAG by adding a linker comprising a maleimide group to form an activated PNAG; b) activating the carrier protein by adding a linker comprising a sulphydryl group to form an activated carrier protein; and c) reacting the activated PNAG and the activated carrier protein to form a PNAG-carrier protein conjugate; or by a process comprising a) activating the PNAG by adding a linker comprising a sulphydryl group to form an activated PNAG; b) activating the carrier protein by adding a linker comprising a maleimide group to form an activated carrier protein; and c) reacting the activated PNAG and the activated carrier protein to form a PNAG-carrier protein conjugate; or by a process comprising a) activating the PNAG by adding a linker comprising a sulphydryl group to form an activated PNAG; b) activating the carrier protein by adding a linker comprising a sulphydryl group to form an activated carrier protein; and c) reacting the activated PNAG and the activated carrier protein to form a PNAG-carrier protein conjugate.
The individual antigens identified in the antigen groups of the invention may be used as carrier proteins for exopolysaccharides, to form a covalent conjugate. Thus the invention provides an immunogenic composition comprising a conjugate of (1) an antigen selected from the first, second, third and fourth antigen groups and (2) a S. aureus exopolysaccharide. The invention also provides an immunogenic composition comprising a conjugate of (1) an antigen selected from the first, second, third and fourth antigen groups and (2) a S.aureus capsular saccharide. Further characteristics of such conjugates are described above. These conjugates may be combined with any of the antigens disclosed herein.
Combinations with non-staphylococcal antigens The individual antigens identified in the antigen groups of the invention may be used in combination with non-staphylococcal antigens, and in particular with antigens from bacteria associated with nosocomial infections. Thus the invention provides an immunogenic composition comprising a combination of:
(1) one or more antigen(s) selected from the first, second, third and fourth antigen groups (as defined above); and
(2) one or more antigen(s) selected from the group consisting of: Clostridium difficile; Pseudomonas aeruginosa; Candida albicans; and extraintestinal pathogenic Escherichia coli.
Further suitable antigens for use in combination with staphylococcal antigens of the invention are listed on pages 33-46 of reference 51. First antigen group clfA
The 'clfA' antigen is annotated as 'clumping factor A'. In the NCTC 8325 strain clfA is SAOUHSC 00812 and has amino acid sequence SEQ ID NO: 1 (GI: 88194572). In the Newman strain it is nwmn_0756 (GI: 151220968).
Useful clfA antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 1 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 1; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 1, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These clfA proteins include variants of SEQ ID NO: 1. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 1. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 1 while retaining at least one epitope of SEQ ID NO: 1. The final 368 C-terminal amino acids of SEQ ID NO: 1 can usefully be omitted. The first 39 N-terminal amino acids of SEQ ID NO: 1 can usefully be omitted. Other fragments omit one or more protein domains.
SEQ ID NO: 224 is a useful fragment of SEQ ID NO: 1 ('CIfA40-559')- This fragments omits the long repetitive region towards the C-terminal of SEQ ID NO: 1. clβ
The 'clfB' antigen is annotated as 'clumping factor B'. hi the NCTC 8325 strain clfB is SAOUHSC 02963 and has amino acid sequence SEQ ID NO: 2 (GI:88196585). In the Newman strain it is nwmn_2529 (GI: 151222741).
Useful clfB antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 2 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%,
65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 2; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
SEQ ID NO: 2, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
90, 100, 150, 200, 250 or more). These clfB proteins include variants of SEQ ID NO: 2. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 2. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ
ID NO: 2 while retaining at least one epitope of SEQ ID NO: 2. The final 40 C-terminal amino acids of SEQ ID NO: 2 can usefully be omitted. The first 44 N-terminal amino acids of SEQ ID NO: 2 can usefully be omitted. Other fragments omit one or more protein domains. ClfB is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc. SEQ ID NO: 163 is a useful fragment of SEQ ID NO: 2 ('CIfB45-552'). This fragment includes the most exposed domain of CIfB and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. Other useful fragments, based on a 3 -domain model of CIfB, include: CIfB45-360 (also known as CLfβ-N12; SEQ ID NO: 196); CIfB2I2-542 (also known as CLfB-N23; SEQ ID NO: 197); and CIfB360-542 (also known as CLfB-N3; SEQ ID NO: 198). coA
The 'coA' antigen is annotated as 'coagulase Coa'. In the NCTC 8325 strain coA is SAOUHSC 00192 and has amino acid sequence SEQ ID NO: 3 (GI:88194002). In the Newman strain it is nwmn_0166 (GI: 151220378). Useful coA antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 3 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 3; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 3, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These coA proteins include variants of SEQ ID NO: 3. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 3. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 3 while retaining at least one epitope of SEQ ID NO: 3. The first 14 N-terminal amino acids of SEQ ED NO: 3 can usefully be omitted. Other fragments omit one or more protein domains. eap
The 'eap' antigen is annotated as 'MHC class II analog protein', hi the NCTC 8325 strain eap is SAOUHSC 02161 and has amino acid sequence SEQ ID NO: 4 (GI: 88195840). hi the Newman strain it is nwmn_1872 (GI: 151222084). Useful eap antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 4 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 4; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 4, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These eap proteins include variants of SEQ ID NO: 4. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 4. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 4 while retaining at least one epitope of SEQ ID NO: 4. The first 17 N-terminal amino acids of SEQ ID NO: 4 can usefully be omitted. Other fragments omit one or more protein domains. ebhA
The 'ebhA' antigen is annotated as 'EbhA'. In the NCTC 8325 strain ebhA is SAOUHSC O 1447 and has amino acid sequence SEQ ID NO: 5 (GI: 88195168).
Useful ebhA antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 5 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 5; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 5, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These ebhA proteins include variants of SEQ ID NO: 5. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 5. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 5 while retaining at least one epitope of SEQ ID NO: 5. The first 39 N- terminal amino acids of SEQ ID NO: 5 can usefully be omitted. Other fragments omit one or more protein domains. ebpS
The 'ebpS' antigen is annotated as 'elastin binding protein EbpS'. In the NCTC 8325 strain ebpS is SAOUHSC 01501 and has amino acid sequence SEQ ID NO: 6 (GI: 88195217). In the Newman strain it is nwmn_1389 (GI:151221601). Useful ebpS antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 6 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 6; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 6, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These ebpS proteins include variants of SEQ ID NO: 6. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 6. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 6 while retaining at least one epitope of SEQ ID NO: 6. Other fragments omit one or more protein domains.
SEQ ID NO: 165 is a useful fragment of SEQ ID NO: 6 (ΕbpSi.i98'). This fragment includes the most exposed domain of EbpS and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. efb
The 'efb' antigen is annotated as 'fϊbrinogen-binding protein truncated'. In the NCTC 8325 strain efb is SAOUHSC Ol 114 and has amino acid sequence SEQ ID NO: 7 (GI: 88194860). In the Newman strain it is nwmn l 069 (GI: 151221281 ). Useful efb antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 7 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 7; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 7, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These efb proteins include variants of SEQ ID NO: 7. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 7. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 7 while retaining at least one epitope of SEQ ID NO: 7. The first 14 N-terminal amino acids of SEQ BD NO: 7 can usefully be omitted. Other fragments omit one or more protein domains. emp
The 'emp' antigen is annotated as 'extracellular matrix and plasma binding protein'. In the NCTC 8325 strain emp is SAOUHSC 00816 and has amino acid sequence SEQ ID NO: 8 (GI: 88194575). In the Newman strain it is nwmn_0758 (GI: 151220970). Useful emp antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 8 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 8; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 8, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These emp proteins include variants of SEQ ID NO: 8. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 8. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 8 while retaining at least one epitope of SEQ ID NO: 8. The first 26 N-terminal amino acids of SEQ ID NO: 8 can usefully be omitted. Other fragments omit one or more protein domains.
SEQ ID NOs: 190, 191, 192 and 193 are useful fragments of SEQ ID NO: 8 (Εmp35-34o\ Εmp27-334\ 'Emp35-334' and 'Emp27-147', respectively). esaC
The 'esaC antigen is annotated as 'esaC. In the NCTC 8325 strain esaC is SAOUHSC 00264 and has amino acid sequence SEQ ID NO: 9 (GI:88194069). Useful esaC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 9 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 9; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 9, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These esaC proteins include variants of SEQ ID NO: 9. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 9. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 9 while retaining at least one epitope of SEQ ID NO: 9. Other fragments omit one or more protein domains. esxA
The 'esxA' antigen is annotated as 'protein'. In the NCTC 8325 strain esxA is SAOUHSC 00257 and has amino acid sequence SEQ ID NO: 10 (GI:88194063). Useful esxA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 10 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 10; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 10, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90 or more). These esxA proteins include variants of SEQ ID NO: 10. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 10. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 10 while retaining at least one epitope of SEQ ID NO: 10. Other fragments omit one or more protein domains. esxB
The 'esxB' antigen is annotated as 'esxB'. In the NCTC 8325 strain esxB is SAOUHSC 00265 and has amino acid sequence SEQ ID NO: 11 (GI:88194070).
Useful esxB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 11 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%,
65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 11; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
SEQ ID NO: 11, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
90, 100 or more). These esxB proteins include variants of SEQ ID NO: 11. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 11. Other preferred fragments lack one or more amino acids
(e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 11 while retaining at least one epitope of SEQ ID NO: 11. Other fragments omit one or more protein domains.
FnBA
The 1FnBA' antigen is annotated as 'fibronectin-binding protein A precursor FnBPA'. In the NCTC 8325 strain FnBA is SAOUHSC 02803 and has amino acid sequence SEQ ID NO: 12 (GL88196438). In the Newman strain it is nwmn_2399 (GI: 151222611). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
Useful FnBA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 12 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 12; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 12, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These FnBA proteins include variants of SEQ ID NO: 12. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 12. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 12 while retaining at least one epitope of SEQ ID NO: 12. The final 37 C- terminal amino acids of SEQ ID NO: 12 can usefully be omitted. Other fragments omit one or more protein domains. FnBA is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc.
SEQ ID NOs: 166 ('FnBAj-511') and 167 ('FnBA5I2-953') are useful fragments of SEQ ID NO: 12. These fragments are more easily used at an industrial scale.
FnBB
The 'FnBB' antigen is annotated as 'fibronectin binding protein B FnBPB'. In the NCTC 8325 strain FnBB is SAOUHSC 02802 and has amino acid sequence SEQ ID NO: 13 (GI:88196437). In the Newman strain it is nwmn_2397 (GI: 151222609).
Useful FnBB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 13 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 13; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 13, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These FnBB proteins include variants of SEQ ID NO: 13. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 13. Other preferred fragments lactone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 13 while retaining at least one epitope of SEQ ID NO: 13. The final 37 C- terminal amino acids of SEQ ID NO: 13 can usefully be omitted. Other fragments omit one or more protein domains.
HIa
The 'HIa' antigen is the 'alpha-hemolysin precursor' also known as 'alpha toxin' or simply 'hemolysin'. hi the NCTC 8325 strain HIa is SAOUHSC Ol 121 and has amino acid sequence SEQ ID NO: 14 (GI: 88194865). In the Newman strain it is nwmn_1073 (GI:151221285). HIa is an important virulence determinant produced by most strains of S.aureus, having pore-forming and haemolytic activity. Anti-Hla antibodies can neutralise the detrimental effects of the toxin in animal models, and HIa is particularly useful for protecting against pneumonia. Useful HIa antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 14 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 14; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 14, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These HIa proteins include variants of SEQ ID NO: 14. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 14. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 14 while retaining at least one epitope of SEQ ID NO: 14. The first 26 N-terminal amino acids of SEQ ID NO: 14 can usefully be omitted (e.g. to give SEQ ID NO: 231). Truncation at the C- terminus can also be used e.g. leaving only 50 amino acids (residues 27-76 of SEQ ID NO: 14) [52]. Other fragments omit one or more protein domains.
HIa' s toxicity can be avoided in compositions of the invention by chemical inactivation (e.g. using formaldehyde, glutaraldehyde or other cross-linking reagents). Instead, however, it is preferred to use mutant forms of HIa which remove its toxic activity while retaining its immunogenicity. Such detoxified mutants are already known in the art. One useful HIa antigen has a mutation at residue 61 of SEQ ID NO: 14, which is residue 35 of the mature antigen (i.e. after omitting the first 26 N-terminal amino acids = residue 35 of SEQ ID NO: 231). Thus residue 61 may not be histidine, and may instead be e.g. He, VaI or preferably Leu. A His-Arg mutation at this position can also be used. For example, SEQ ID NO: 150 is the mature mutant Hla-H35L sequence (i.e. SEQ ID NO: 231 with a H35L mutation) and a useful HIa antigen comprises SEQ ID NO: 150. Another useful mutation replaces a long loop with a short sequence e.g. to replace the 39mer at residues 136-174 of SEQ ID NO: 14 with a tetramer such as PSGS (SEQ ID NO: 225), as in SEQ ID NO: 189 (which also includes the H35L mutation) and SEQ ID NO: 216 (which does not include the H35L mutation). Another useful mutation replaces residue YlOl e.g. with a leucine (SEQ ID NO: 242). Another useful mutation replaces residue D 152 e.g. with a leucine (SEQ ID NO: 243). Another useful mutant replaces residues H35 and YlOl e.g. with a leucine (SEQ ID NO: 244). Another useful mutant replaces residues H35 and D152 e.g. with a leucine (SEQ ID NO: 245).
Further useful HIa antigens are disclosed in references 53 and 54.
SEQ ID NOs: 160, 161 & 194 are three useful fragments of SEQ ID NO: 14 ('HIa27-76', 'HIa27-89' and 'HIa27-79', respectively). SEQ ID NOs: 158, 159 and 195 are the corresponding fragments from SEQ ID NO: 150.
One useful HIa sequence is SEQ ID NO: 232, which was used in the examples. It has a N-terminal Met, then an Ala-Ser dipeptide from the expression vector, then SEQ ID NO: 150 (from NCTC8325 strain). It is encoded by SEQ ID NO: 233. MgB
The 'hlgB' antigen is annotated as 'leukocidin f subunit precursor HIgB'. In the NCTC 8325 strain hlgB is SAOUHSC 02710 and has amino acid sequence SEQ ID NO: 15 (GI:88196350).
Useful hlgB antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 15 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 15; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 15, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These hlgB proteins include variants of SEQ ID NO: 15. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 15. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 15 while retaining at least one epitope of SEQ ID NO: 15. The first 26 N-terminal amino acids of SEQ ID NO: 15 can usefully be omitted. Other fragments omit one or more protein domains.
MgC The 'hlgC antigen is annotated as 'leukocidin s subunit precursor HIgC. In the NCTC 8325 strain hlgC is SAOUHSC_02709 and has amino acid sequence SEQ ID NO: 16 (GI: 88196349).
Useful hlgC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 16 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 16; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 16, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These hlgC proteins include variants of SEQ ID NO: 16. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 16. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 16 while retaining at least one epitope of SEQ ID NO: 16. The first 29 N-terminal amino acids of SEQ ID NO: 16 can usefully be omitted. Other fragments omit one or more protein domains. isdA
The 'isdA' antigen is annotated as 'IsdA protein'. In the NCTC 8325 strain isdA is SAOUHSC O 1081 and has amino acid sequence SEQ ID NO: 17 (GI: 88194829). In the Newman strain it is nwmn_1041 (GI: 151221253).
Useful isdA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 17 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 17; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 17, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These isdA proteins include variants of SEQ ID NO: 17. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 17. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 17 while retaining at least one epitope of SEQ ID NO: 17. The final 38 C-terminal amino acids of SEQ ID NO: 17 can usefully be omitted. The first 46 N-terminal amino acids of SEQ ID NO: 17 can usefully be omitted. Truncation to exclude the C-terminal 38mer of SEQ ID NO: 17 (beginning with the LPKTG motif) is also useful. Other fragments omit one or more protein domains.
SEQ ID NO: 157 is a useful fragment of SEQ ID NO: 17 (amino acids 40-184 of SEQ ID NO: 17; 'ISdA4O-I84') which includes the natural protein's heme binding site and includes the antigen's most exposed domain. It also reduces the antigen's similarity with human proteins. Other useful fragments are disclosed in references 55 and 56. IsdA does not adsorb well to aluminium hydroxide adjuvants, so IsdA present in a composition may me unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate.
Anti-IsdA antibodies protect mice against S. aureus abscess formation and lethal challenge [57]. isdB
The 'isdB' antigen is annotated as 'neurofilament protein isdB'. In the NCTC 8325 strain isdB is SAOUHSC 01079 and has amino acid sequence SEQ ID NO: 18 (GL88194828). IsdB has been proposed for use as a vaccine antigen on its own [2], but this may not prevent pneumonia.
Useful isdB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
ID NO: 18 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%,
65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 18; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 18, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These isdB proteins include variants of SEQ ID NO: 18. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 18. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 18 while retaining at least one epitope of SEQ ID NO: 18. The final 36 C-terminal amino acids of SEQ ID NO: 18 can usefully be omitted. The first 40 N-terminal amino acids of SEQ ID NO: 18 can usefully be omitted. Other fragments omit one or more protein domains. Useful fragments of IsdB are disclosed in references 56 and 58 e.g. lacking 37 internal amino acids of SEQ ID 18.
Anti-IsdB antibodies protect mice against S.aureus abscess formation and lethal challenge [57]. In some embodiments, compositions of the invention do not include an isdB antigen. isdC
The 'isdC antigen is annotated as 'protein'. In the NCTC 8325 strain isdC is SAOUHSC O 1082 and has amino acid sequence SEQ ID NO: 19 (GI: 88194830).
Useful isdC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 19 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 19; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 19, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These isdC proteins include variants of SEQ ID NO: 19. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 19. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 19 while retaining at least one epitope of SEQ ID NO: 19. The final 39 C-terminal amino acids of SEQ ID NO: 19 can usefully be omitted. The first 28 N-terminal amino acids of SEQ ID NO: 19 can usefully be omitted. Other fragments omit one or more protein domains. Useful fragments of IsdB are disclosed in reference 56.
Reference 59 discloses antigens which usefully include epitopes from both IsdB and IsdH. isdG
The 'isdG' antigen is annotated as 'heme-degrading monooxygenase IsdG'. In the NCTC 8325 strain isdG is SAOUHSC 01089 and has amino acid sequence SEQ ID NO: 20 (GI: 88194836).
Useful isdG antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
ID NO: 20 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 20; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 20, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These isdG proteins include variants of SEQ ID NO: 20. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 20. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 20 while retaining at least one epitope of SEQ ID NO: 20. Other fragments omit one or more protein domains. isdH The 'isdH' antigen is annotated as 'isdH'. In the NCTC 8325 strain isdH is SAOUHSC Ol 843 and has amino acid sequence SEQ ID NO: 21 (GI:88195542). In the Newman strain it is nwmn_1624 (GI: 151221836). It has also been known as HarA.
Useful isdH antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 21 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 21; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 21, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These isdH proteins include variants of SEQ ID NO: 21. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 21. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 21 while retaining at least one epitope of SEQ ID NO: 21. The final 35 C-terminal amino acids of SEQ ID NO: 21 can usefully be omitted. The first 40 N-terminal amino acids of SEQ ID NO: 21 can usefully be omitted. Other fragments omit one or more protein domains. Reference 59 discloses antigens which usefully include epitopes from both IsdB and IsdH. isdl
The 'isdl' antigen is annotated as 'heme-degrading monooxygenase Isdl'. In the NCTC 8325 strain isdl is SAOUHSC OO 130 and has amino acid sequence SEQ ID NO: 22 (GI: 88193943).
Useful isdl antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 22 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%,
65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 22; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of
SEQ ID NO: 22, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80,
90, 100 or more). These isdl proteins include variants of SEQ ID NO: 22. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 22. Other preferred fragments lack one or more amino acids
(e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 22 while retaining at least one epitope of SEQ ID NO: 22. Other fragments omit one or more protein domains. lukD The 'lukD' antigen is annotated as 'leukotoxin LukD'. In the NCTC 8325 strain lukD is SAOUHSC 01954 and has amino acid sequence SEQ ID NO: 23 (GI:88195647). hi the Newman strain it is nwmn l 718 (GI: 151221930).
Useful lukD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 23 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 23; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 23, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These lukD proteins include variants of SEQ ID NO: 23. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 23. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 23 while retaining at least one epitope of SEQ ID NO: 23. The final 43 C-terminal amino acids of SEQ ID NO: 23 can usefully be omitted. The first 26 N-terminal amino acids of SEQ ID NO: 23 can usefully be omitted. Other fragments omit one or more protein domains. lukE
The 'lukE' antigen is annotated as 'leukotoxin LukE'. In the NCTC 8325 strain lukE is SAOUHSCJ) 1955 and has amino acid sequence SEQ ID NO: 24 (GI: 88195648).
Useful lukE antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 24 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 24; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 24, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These lukE proteins include variants of SEQ ID NO: 24. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 24. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 24 while retaining at least one epitope of SEQ ID NO: 24. Other fragments omit one or more protein domains. lukF The 'lukF' antigen is annotated as 'Leukocidin/Hemolysin toxin family LukF'. In the NCTC 8325 strain lukF is SAOUHSC 02241 and has amino acid sequence SEQ ID NO: 25 (GI:88195914). Useful lukF antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 25 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 25; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 25, wherein 'n? is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These lukF proteins include variants of SEQ ID NO: 25. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 25. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 25 while retaining at least one epitope of SEQ ID NO: 25. Other fragments omit one or more protein domains. lukS
The 'lukS' antigen is annotated as 'probable leukocidin S subunit LukS'. In the NCTC 8325 strain lukS is SAOUHSC_02243 and has amino acid sequence SEQ ID NO: 26 (GL88195915). In the Newman strain it is nwmn_1928 (GI: 151222140).
Useful lukS antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 26 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 26; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 26, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These lukS proteins include variants of SEQ ID NO: 26. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 26. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 26 while retaining at least one epitope of SEQ ID NO: 26. The first 22 N-terminal amino acids of SEQ ID NO: 26 can usefully be omitted. Other fragments omit one or more protein domains. nuc
The 'nuc' antigen is annotated as 'thermonuclease precursor'. In the NCTC 8325 strain nuc is
SAOUHSC 01316 and has amino acid sequence SEQ ID NO: 27 (GI:88195046). Useful nuc antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 27 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 27; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 27, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These nuc proteins include variants of SEQ ID NO: 27. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 27. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 27 while retaining at least one epitope of SEQ ID NO: 27. The final 39 C-terminal amino acids of SEQ ID NO: 27 can usefully be omitted. The first 19 N-terminal amino acids of SEQ ID NO: 27 can usefully be omitted. Other fragments omit one or more protein domains. sasA
The 'sasA' antigen is annotated as 'SasA'. In the NCTC 8325 strain sasA is SAOUHSC 02990 and has amino acid sequence SEQ ID NO: 28 (GI:88196609).
Useful sasA antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 28 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 28; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 28, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sasA proteins include variants of SEQ ID NO: 28. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 28. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 28 while retaining at least one epitope of SEQ ID NO: 28. The final 43 C-terminal amino acids of SEQ ID NO: 28 can usefully be omitted. The first 90 N-terminal amino acids of SEQ ID NO: 28 can usefully be omitted. Other fragments omit one or more protein domains. sasB
The 'sasB' antigen is annotated as 'frntB protein; SasB'. hi the NCTC 8325 strain sasB is SAOUHSC 02404 and has amino acid sequence SEQ ID NO: 29 (GI: 88196065).
Useful sasB antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 29 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 29; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 29, wherein W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sasB proteins include variants of SEQ ID NO: 29. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 29. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 29 while retaining at least one epitope of SEQ ID NO: 29. The final 39 C-terminal amino acids of SEQ ID NO: 29 can usefully be omitted. The first 38 N-terminal amino acids of SEQ ID NO: 29 can usefully be omitted. Other fragments omit one or more protein domains. sasC
The 'sasC antigen is annotated as 'Mrp protein; SasC. In the NCTC 8325 strain sasC is SAOUHSC 01873 and has amino acid sequence SEQ ID NO: 30 (GI:88195570).
Useful sasC antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 30 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 30; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 30, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sasC proteins include variants of SEQ ID NO: 30. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 30. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 30 while retaining at least one epitope of SEQ ID NO: 30. The final 36 C-terminal amino acids of SEQ ID NO: 30 can usefully be omitted. The first 37 N-terminal amino acids of SEQ ID NO: 30 can usefully be omitted. Other fragments omit one or more protein domains. sasD
The 'sasD' antigen is annotated as 'SasD protein'. In the NCTC 8325 strain sasD is
SAOUHSC 00094 and has amino acid sequence SEQ ID NO: 31 (GL88193909).
Useful sasD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 31 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 31; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 31, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These sasD proteins include variants of SEQ ID NO: 31. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 31. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 31 while retaining at least one epitope of SEQ ID NO: 31. The first 28 N-terminal amino acids of SEQ ID NO: 31 can usefully be omitted. Other fragments omit one or more protein domains. sasF
The 'sasF antigen is annotated as 'sasF protein'. In the NCTC 8325 strain sasF is SAOUHSC 02982 and has amino acid sequence SEQ ID NO: 32 (GI:88196601).
Useful sasF antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ
ED NO: 32 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 32; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 32, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sasF proteins include variants of SEQ ID NO: 32. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 32. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 32 while retaining at least one epitope of SEQ ID NO: 32. The final 39 C-terminal amino acids of SEQ ID NO: 32 can usefully be omitted. The first 37 N-terminal amino acids of SEQ ID NO: 32 can usefully be omitted. Other fragments omit one or more protein domains. sdrC The 'sdrC antigen is annotated as 'sdrC protein1, hi the NCTC 8325 strain sdrC is SAOUHSC 00544 and has amino acid sequence SEQ ID NO: 33 (GI: 88194324).
Useful sdrC antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 33 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 33; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 33, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sdrC proteins include variants of SEQ ID NO: 33. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 33. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 33 while retaining at least one epitope of SEQ ID NO: 33. The final 38 C-terminal amino acids of SEQ ID NO: 33 can usefully be omitted. The first 50 N-terminal amino acids of SEQ ID NO: 33 can usefully be omitted. Other fragments omit one or more protein domains. SdrC is naturally a long protein and so the use of fragments is helpful e.g. for purification, handling, fusion, expression, etc.
SEQ ID NO: 164 is a useful fragment of SEQ ID NO: 33 ('SdrC5i-5i8')- This fragment includes the most exposed domain of SdrC and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. sdrD The 'sdrD' antigen is annotated as 'sdrD protein'. In the NCTC 8325 strain sdrD is SAOUHSC 00545 and has amino acid sequence SEQ ID NO: 34 (GI: 88194325).
Useful sdrD antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 34 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 34; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 34, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sdrD proteins include variants of SEQ ID NO: 34. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 34. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 34 while retaining at least one epitope of SEQ ID NO: 34. The final 38 C-terminal amino acids of SEQ ID NO: 34 can usefully be omitted. The first 52 N-terminal amino acids of SEQ ID NO: 34 can usefully be omitted. Other fragments omit one or more protein domains. SdrD is naturally a long protein and so the use of fragments is very helpful e.g. for purification, handling, fusion, expression, etc. SEQ ID NO: 156 is a useful fragment of SEQ ID NO: 34 ('SdTD53-592'). This fragment includes the most exposed domain of SdrD and is more easily used at an industrial scale. It also reduces the antigen's similarity with human proteins. Another useful fragment, with the same C-terminus residue, is SdTD394-592 (also known as SdrD-N3; SEQ ID NO: 199). Another useful fragment is SEQ ID NO: 236 (amino acids 593-1123 of SEQ ID NO: 34), referred to herein as 'SdrDCnaB'. sdrE2
The 'sdrE2' antigen is annotated as 'Ser-Asp rich fibrinogen/bone sialoprotein-binding protein SdrE'. In the Newman strain sdrE2 is NWMN 0525 and has amino acid sequence SEQ ID NO: 35 (GL151220737).
Useful sdrE2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 35 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 35; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 35, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These sdrE2 proteins include variants of SEQ ID NO: 35. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 35. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 35 while retaining at least one epitope of SEQ YD NO: 35. The final 38 C- terminal amino acids of SEQ ID NO: 35 can usefully be omitted. The first 52 N-terminal amino acids of SEQ ID NO: 35 can usefully be omitted. Other fragments omit one or more protein domains.
SdrE2 is naturally a long protein and so the use of fragments is very helpful e.g. for purification, handling, fusion, expression, etc.
SEQ ID NO: 155 is a useful fragment of SEQ ID NO: 35 ('SdrE53-632'). This fragment includes the most exposed domain of SdrE2 and is more easily used at an industrial scale. It also reduces the antigen' s similarity with human proteins. spa
The 'spa' antigen is annotated as 'protein A' or 'SpA'. In the NCTC 8325 strain spa is SAOUHSC 00069 and has amino acid sequence SEQ ID NO: 36 (GI:88193885). In the Newman strain it is nwmn_0055 (GI: 151220267). All S.aureus strains express the structural gene for spa, a well characterized virulence factor whose cell wall-anchored surface protein product has five highly homologous immunoglobulin binding domains designated E, D, A, B, and C [60]. These domains display -80% identity at the amino acid level, are 56 to 61 residues in length, and are organized as tandem repeats [61]. SpA is synthesized as a precursor protein with an N-terminal signal peptide and a C-terminal sorting signal [62,63]. Cell wall-anchored spa is displayed in great abundance on the staphylococcal surface [64,65]. Each of its immunoglobulin binding domains is composed of anti- parallel α-helices that assemble into a three helix bundle and can bind the Fc domain of immunoglobulin G (IgG) [66,67], the VH3 heavy chain (Fab) of IgM {i.e. the B cell receptor) [68], the von Willebrand factor at its Al domain [69] and/or the TNF-α receptor I (TNFRI) [70], which is displayed on surfaces of airway epithelia. Useful spa antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 36 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 36; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 36, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These spa proteins include variants of SEQ ID NO: 36. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 36. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 36 while retaining at least one epitope of SEQ ID NO: 36. The final 35 C-terminal amino acids of SEQ ID NO: 36 can usefully be omitted. The first 36 N-terminal amino acids of SEQ ID NO: 36 can usefully be omitted. Other fragments omit one or more protein domains. Reference 71 suggests that individual IgG-binding domains might be useful immunogens, alone or in combination.
SEQ ID NO: 162 is a useful fragment of SEQ ID NO: 36 ('Spa37-325'). This fragment contains all the five SpA Ig-binding domains (which are naturally arranged from N- to C-terminus in the order E, D, A, B, C) and includes the most exposed domain of SpA. It also reduces the antigen's similarity with human proteins. Other useful fragments may omit 1, 2, 3 or 4 of the natural A, B, C, D and/or E domains to prevent the excessive B cell expansion and then apoptosis which might occur if spa functions as a B cell superantigen. As reported in reference 71, other useful fragments may include only 1, 2, 3 or 4 of the natural A, B, C, D and/or E domains e.g. comprise only the SpA(A) domain but not B to E, or comprise only the SpA(D) domain but not A, B, C or E, etc. Thus a spa antigen useful with the invention may include 1, 2, 3, 4 or 5 IgG-binding domains, but ideally has 4 or fewer If an antigen includes only one type of spa domain (e.g. only the Spa(A) or SpA(D) domain), it may include more than one copy of this domain e.g. multiple SpA(D) domains in a single polypeptide chain.
An individual domain within the antigen may be mutated at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids relative to SEQ ID NO: 36 (e.g. see ref. 71, disclosing mutations at residues 3 and/or 24 of domain D, at residue 46 and/or 53 of domain A, etc.). Such mutants should not remove the antigen's ability to elicit an antibody that recognises SEQ ID NO: 36, but may remove the antigen's binding to IgG and/or other human proteins (such as human blood proteins).
In certain aspects a spa antigen includes a substitution at (a) one or more amino acid substitution in an IgG Fc binding sub-domain of SpA domain A, B, C, D and/or E that disrupts or decreases binding to IgG Fc, and (b) one or more amino acid substitution in a VH3 binding sub-domain of SpA domain A, B, C, D, and/or E that disrupts or decreases binding to VH3. In certain embodiments, a variant SpA comprises at least or at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more variant SpA domain D peptides.
Second antigen group staOOl
The 'staOOl1 antigen is annotated as '5'-nucleotidase family protein', hi the NCTC 8325 strain staOOl is SAOUHSC 00025 and has amino acid sequence SEQ ID NO: 37 (GI:88193846). hi the Newman strain it is nwmn_0022 (GI: 151220234). It has also been referred to as AdsA and SasH and SA0024.
Useful staOOl antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 37 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 37; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 37, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staOOl proteins include variants of SEQ ID NO: 37. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 37. Other preferred fragments lactone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 37 while retaining at least one epitope of SEQ ID NO: 37. The final 34 C- teπninal amino acids of SEQ ID NO: 37 can usefully be omitted. The first 38 N-terminal amino acids of SEQ ID NO: 37 can usefully be omitted. Other fragments omit one or more protein domains. sta002
The (sta002' antigen is annotated as 'lipoprotein', hi the NCTC 8325 strain sta002 is SAOUHSC_00356 and has amino acid sequence SEQ ID NO: 38 (GL88194155). hi the Newman strain it is nwmn_0364 (GI: 151220576). Useful sta002 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 38 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 38; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 38, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These sta002 proteins include variants of SEQ ID NO: 38. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 38. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 38 while retaining at least one epitope of SEQ ID NO: 38. The first 18 N-terminal amino acids of SEQ ID NO: 38 can usefully be omitted. Other fragments omit one or more protein domains. SEQ ID NOs: 153 ('sta002i9-i87') and 154 ('sta002i9-124') are two useful fragments of SEQ ID NO: 38 which reduce the antigen's similarity with human proteins. sta003
The 'staOO3' antigen is annotated as 'surface protein', hi the NCTC 8325 strain staOO3 is SAOUHSC 00400 and has amino acid sequence SEQ ID NO: 39 (GI: 88194195). In the Newman strain it is nwmn_0401 (GI: 151220613).
Useful staOO3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 39 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 39; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 39, wherein W is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta003 proteins include variants of SEQ ID NO: 39. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 39. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 39 while retaining at least one epitope of SEQ ID NO: 39. The first 32 N- terminal amino acids of SEQ ID NO: 39 can usefully be omitted. Other fragments omit one or more protein domains. sta004
The 'staOO4' antigen is annotated as 'Siderophore binding protein FatB'. In the NCTC 8325 strain staOO4 is SAOUHSC_00749 and has amino acid sequence SEQ ID NO: 40 (GI: 88194514). In the Newman strain it is nwmn_0705 (GI: 151220917).
Useful staOO4 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 40 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 40; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 40, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta004 proteins include variants of SEQ ID NO: 40. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 40. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 40 while retaining at least one epitope of SEQ ID NO: 40. The first 18 N- terminal amino acids of SEQ ID NO: 40 can usefully be omitted. Other fragments omit one or more protein domains. sta005
The 'staOO5' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staOO5 is SAOUHSC Ol 127 and has amino acid sequence SEQ ID NO: 41 (GI: 88194870). In the Newman strain it is nwmn_1077 (GI:151221289).
Useful sta005 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 41 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 41; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 41, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staOO5 proteins include variants of SEQ ID NO: 41. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 41. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 41 while retaining at least one epitope of SEQ ID NO: 41. The first 18 N-terminal amino acids of SEQ ID NO: 41 can usefully be omitted. Other fragments omit one or more protein domains. staOOό
The 'staOOό' antigen is annotated as 'ferrichrome-binding protein1, and has also been referred to as 'FhuD2' in the literature [72]. In the NCTC 8325 strain staOOό is SAOUHSC 02554 and has amino acid sequence SEQ ID NO: 42 (GL88196199). In the Newman strain it is nwmn_2185 (GI: 151222397).
Useful staOOό antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 42 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 42; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 42, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staOOό proteins include variants of SEQ ID NO: 42. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 42. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 42 while retaining at least one epitope of SEQ ID NO: 42. The first 17 N-terminal amino acids of SEQ ID NO: 42 can usefully be omitted (to provide SEQ ID NO: 246). Other fragments omit one or more protein domains. Mutant forms of staOOό are reported in reference 73. A staOOό antigen may be lipidated e.g. with an acylated N-terminus cysteine. One useful staOOό sequence is SEQ ID NO: 248, which has a Met-Ala-Ser- sequence at the N-terminus. sta007
The 'sta007' antigen is annotated as 'secretory antigen precursor', hi the NCTC 8325 strain sta007 is SAOUHSC 02571 and has amino acid sequence SEQ ID NO: 43 (GI:88196215). hi the Newman strain it is nwmn_2199 (GI: 151222411). Proteomic analysis has revealed that this protein is secreted or surface-exposed. Useful sta007 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 43 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 43; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 43, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta007 proteins include variants of SEQ ID NO: 43. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 43. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ED NO: 43 while retaining at least one epitope of SEQ DD NO: 43. The first 27 N- terminal amino acids of SEQ DD NO: 43 can usefully be omitted. Other fragments omit one or more protein domains. staOO8
The 'staOO8' antigen is annotated as 'lipoprotein', hi the NCTC 8325 strain staOO8 is SAOUHSC 02650 and has amino acid sequence SEQ ID NO: 44 (GI:88196290). hi the Newman strain it is nwmn_2270 (GI: 151222482).
Useful staOO8 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 44 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 44; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 44, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staOO8 proteins include variants of SEQ ID NO: 44. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 44. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ DD NO: 44 while retaining at least one epitope of SEQ ID NO: 44. The first 17 N-terminal amino acids of SEQ ID NO: 44 can usefully be omitted. Other fragments omit one or more protein domains. sta.009
The 'sta009' antigen is annotated as 'immunoglobulin G-binding protein Sbi'. In the NCTC 8325 strain sta009 is SAOUHSC 02706 and has amino acid sequence SEQ ID NO: 45 (GI:88196346). In the Newman strain it is nwmn_2317 (GI: 151222529). Useful sta009 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 45 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 45; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 45, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta009 proteins include variants of SEQ ID NO: 45. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 45. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 45 while retaining at least one epitope of SEQ ID NO: 45. The first 29 N- terminal amino acids of SEQ ID NO: 45 can usefully be omitted. Other fragments omit one or more protein domains. staOlO
The 'staOlO' antigen is annotated as 'immunodominant antigen A'. In the NCTC 8325 strain staOlO is SAOUHSC_02887 and has amino acid sequence SEQ ID NO: 46 (GI: 88196515). In the Newman strain it is nwmn_2469 (GI: 151222681). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
Useful staOlO antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 46 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 46; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 46, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staOlO proteins include variants of SEQ ID NO: 46. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 46. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 46 while retaining at least one epitope of SEQ ID NO: 46. The first 29 N-terminal amino acids of SEQ ED NO: 46 can usefully be omitted. Other fragments omit one or more protein domains. staOll
The 'staOl l' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staOl l is SAOUHSC_00052 and has amino acid sequence SEQ ID NO: 47 (GL88193872). Useful staOl l antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 47 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 47; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 47, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staOl l proteins include variants of SEQ ID NO: 47. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 47. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 47 while retaining at least one epitope of SEQ ID NO: 47. The first 23 N-terminal amino acids of SEQ ID NO: 47 can usefully be omitted (to provide SEQ ID NO: 247). Other fragments omit one or more protein domains. A staOl l antigen may be lipidated e.g. with an acylated N-terminus cysteine. One useful staOl l sequence is SEQ ID NO: 249, which has a N-terminus methionine. Variant forms of SEQ ID NO: 47 which may be used as or for preparing staOl 1 antigens include, but are not limited to, SEQ ID NOs: 213, 214 and 215 with various Ile/Val/Leu substitutions.
StaOl l can exist as a monomer or an oligomer, with Ca+* ions favouring oligomerisation. The invention can use monomers and/or oligomers of StaOl 1. staO12 The 'staO12' antigen is annotated as 'protein with leader'. In the NCTC 8325 strain staO12 is SAOUHSC_00106 and has amino acid sequence SEQ ID NO: 48 (GI:88193919).
Useful staOl 2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 48 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 48; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 48, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO12 proteins include variants of SEQ ID NO: 48. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 48. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 48 while retaining at least one epitope of SEQ ID NO: 48. The first 21 N- terminal amino acids of SEQ ID NO: 48 can usefully be omitted. Other fragments omit one or more protein domains. staO13
The ?sta013' antigen is annotated as 'poly-gamnia-glutamate capsule biosynthesis protein', hi the NCTC 8325 strain staO13 is SAOUHSC 00107 and has amino acid sequence SEQ ID NO: 49 (GI: 88193920). Useful staO13 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 49 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 49; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 49, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO13 proteins include variants of SEQ ID NO: 49. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 49. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 49 while retaining at least one epitope of SEQ ID NO: 49. Other fragments omit one or more protein domains. staO14
The 'staO14' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO14 is
SAOUHSC_00137 and has amino acid sequence SEQ ED NO: 50 (GI:88193950).
Useful staO14 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 50 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 50; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 50, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These staO14 proteins include variants of SEQ ID NO: 50. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 50. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 50 while retaining at least one epitope of SEQ ID NO: 50. The first 17 N- terminal amino acids of SEQ ID NO: 50 can usefully be omitted. Other fragments omit one or more protein domains. staO15
The 'staO15' antigen is annotated as 'extracellular solute-binding protein; RGD containing lipoprotein'. In the NCTC 8325 strain staO15 is SAOUHSC OO 170 and has amino acid sequence SEQ ID NO: 51 (GI: 88193980). Useful staO15 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 51 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 51; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 51, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO15 proteins include variants of SEQ ID NO: 51. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 51. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 51 while retaining at least one epitope of SEQ ID NO: 51. The first 18 N- terminal amino acids of SEQ ID NO: 51 can usefully be omitted. Other fragments omit one or more protein domains. staOlό
The 'staOlό' antigen is annotated as 'gamma-glutamyltranspeptidase'. In the NCTC 8325 strain staOlό is SAOUHSC 00171 and has amino acid sequence SEQ ID NO: 52 (GI:88193981).
Useful staOlό antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 52 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 52; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 52, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staOlό proteins include variants of SEQ ID NO: 52. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 52. Other preferred fragments lact¬ one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 52 while retaining at least one epitope of SEQ ID NO: 52. Other fragments omit one or more protein domains. sta017
The 'staO17' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO17 is SAOUHSC OO 186 and has amino acid sequence SEQ ID NO: 53 (GI: 88193996).
Useful staO17 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 53 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 53; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 53, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO17 proteins include variants of SEQ ID NO: 53. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 53. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 53 while retaining at least one epitope of SEQ ID NO: 53. The first 17 N- terminal amino acids of SEQ ID NO: 53 can usefully be omitted. Other fragments omit one or more protein domains. staO18
The 'staO18' antigen is annotated as 'extracellular solute-binding protein'. In the NCTC 8325 strain staO18 is SAOUHSC 00201 and has amino acid sequence SEQ DD NO: 54 (GL88194011).
Useful staO18 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 54 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 54; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 54, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO18 proteins include variants of SEQ ID NO: 54. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 54. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 54 while retaining at least one epitope of SEQ ID NO: 54. Other fragments omit one or more protein domains. staO19
The 'staO19' antigen is annotated as 'peptidoglycan hydrolase'. In the NCTC 8325 strain staO19 is SAOUHSC_00248 and has amino acid sequence SEQ ID NO: 55 (GI:88194055). In the Newman strain it is nwmn_0210 (GI: 151220422).
Useful staO19 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 55 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 55; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 55, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO19 proteins include variants of SEQ ID NO: 55. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 55. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 55 while retaining at least one epitope of SEQ ID NO: 55. The first 25 N- terminal amino acids of SEQ DD NO: 55 can usefully be omitted. Other fragments omit one or more protein domains. Useful fragments are SEQ DD NOs: 228 and 229.
StaO19 does not adsorb well to aluminium hydroxide adjuvants, so StaO19 present in a composition may me unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate. staO2O
The 'sta020' antigen is annotated as 'exported protein'. In the NCTC 8325 strain sta020 is SAOUHSC_00253 and has amino acid sequence SEQ ID NO: 56 (GI:88194059).
Useful sta020 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 56 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 56; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 56, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta020 proteins include variants of SEQ ID NO: 56. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 56. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 56 while retaining at least one epitope of SEQ ID NO: 56. The first 30 N- terminal amino acids of SEQ ID NO: 56 can usefully be omitted. Other fragments omit one or more protein domains. staO21
The 'staO21' antigen is annotated as 'secretory antigen SsaA-like protein1. In the NCTC 8325 strain staO21 is SAOUHSC 00256 and has amino acid sequence SEQ ID NO: 57 (GI: 88194062).
Useful staO21 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 57 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 57; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 57, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These staO21 proteins include variants of SEQ ID NO: 57. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 57. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 57 while retaining at least one epitope of SEQ ID NO: 57. The first 24 N- terminal amino acids of SEQ ID NO: 57 can usefully be omitted. Other fragments omit one or more protein domains. staO22
The 'staO22' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO22 is
SAOUHSC_00279 and has amino acid sequence SEQ ID NO: 58 (GI:88194083).
Useful staO22 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 58 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 58; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 58, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These staO22 proteins include variants of SEQ ID NO: 58. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 58. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 58 while retaining at least one epitope of SEQ ID NO: 58. The first 17 N-terminal amino acids of SEQ ID NO: 58 can usefully be omitted. Other fragments omit one or more protein domains. staO23 The 'staO23' antigen is annotated as ^'-nucleotidase; lipoprotein e(P4) family1. In the NCTC 8325 strain staO23 is SAOUHSC 00284 and has amino acid sequence SEQ ID NO: 59 (GI:88194087).
Useful staO23 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 59 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 59; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 59, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO23 proteins include variants of SEQ ID NO: 59. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 59. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 59 while retaining at least one epitope of SEQ ID NO: 59. The first 31 N- terminal amino acids of SEQ ID NO: 59 can usefully be omitted. Other fragments omit one or more protein domains. staO24 The 'staO24' antigen is annotated as 'lipase precursor'. In the NCTC 8325 strain staO24 is SAOUHSC 00300 and has amino acid sequence SEQ ID NO: 60 (GI:88194101).
Useful staO24 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 60 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 60; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 60, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO24 proteins include variants of SEQ ID NO: 60. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 60. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 60 while retaining at least one epitope of SEQ ID NO: 60. The first 37 N- terminal amino acids of SEQ ID NO: 60 can usefully be omitted. Other fragments omit one or more protein domains. staO25
The 'staO25' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO25 is SAOUHSC_00362 and has amino acid sequence SEQ ID NO: 61 (GI: 88194160).
Useful staO25 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ED NO: 61 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 61; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 61, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO25 proteins include variants of SEQ ID NO: 61. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 61. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 61 while retaining at least one epitope of SEQ ED NO: 61. The first 19 N-terminal amino acids of SEQ ID NO: 61 can usefully be omitted. Other fragments omit one or more protein domains. staO26
The 'staO26' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO26 is
SAOUHSC_00404 and has amino acid sequence SEQ ID NO: 62 (GI:88194198). Useful staO26 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 62 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 62; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 62, wherein ?n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO26 proteins include variants of SEQ ID NO: 62. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 62. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 62 while retaining at least one epitope of SEQ ID NO: 62. The first 22 N- terminal amino acids of SEQ DD NO: 62 can usefully be omitted. Other fragments omit one or more protein domains. staO27
The 'staO27' antigen is annotated as 'probable lipase'. In the NCTC 8325 strain staO27 is
SAOUHSC_00661 and has amino acid sequence SEQ ID NO: 63 (GL88194426). Useful staO27 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 63 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 63; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 63, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO27 proteins include variants of SEQ ID NO: 63. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 63. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 63 while retaining at least one epitope of SEQ ID NO: 63. The first 23 N- terminal amino acids of SEQ ID NO: 63 can usefully be omitted. Other fragments omit one or more protein domains. staO28
The 'staO28' antigen is annotated as 'secretory antigen SsaA-like protein'. In the NCTC 8325 strain staO28 is SAOUHSC 00671 and has amino acid sequence SEQ ID NO: 64 (GI:88194436). In the Newman strain it is nwmn_0634 (GI: 151220846). Useful staO28 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 64 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 64; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 64, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO28 proteins include variants of SEQ ID NO: 64. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 64. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 64 while retaining at least one epitope of SEQ ID NO: 64. The first 25 N- terminal amino acids of SEQ ID NO: 64 can usefully be omitted. Other fragments omit one or more protein domains. staO29
The 'staO29' antigen is annotated as 'ferrichrome binding protein'. In the NCTC 8325 strain staO29 is
SAOUHSC 00754 and has amino acid sequence SEQ ID NO: 65 (GI: 88194518). Useful staO29 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 65 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 65; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 65, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO29 proteins include variants of SEQ ID NO: 65. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 65. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 65 while retaining at least one epitope of SEQ ID NO: 65. The final 25 C- terminal amino acids of SEQ ID NO: 65 can usefully be omitted. The first 19 N-terminal amino acids of SEQ DD NO: 65 can usefully be omitted. Other fragments omit one or more protein domains. sta030
The 'staO3O' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO3O is SAOUHSC_00808 and has amino acid sequence SEQ ID NO: 66 (GI: 88194568).
Useful staO3O antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 66 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 66; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 66, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO3O proteins include variants of SEQ ID NO: 66. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 66. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 66 while retaining at least one epitope of SEQ ID NO: 66. The first 17 N-terminal amino acids of SEQ ID NO: 66 can usefully be omitted. Other fragments omit one or more protein domains. staO31 The 'staO31' antigen is annotated as '5 -nucleotidase family protein'. In the NCTC 8325 strain staO31 is SAOUHSC 00860 and has amino acid sequence SEQ ID NO: 67 (GI:88194617).
Useful staO31 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 67 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 67; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 67, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO31 proteins include variants of SEQ ID NO: 67. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 67. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 67 while retaining at least one epitope of SEQ ID NO: 67. Other fragments omit one or more protein domains. staO32
The 'staO32' antigen is annotated as 'serine protease HtrA1. In the NCTC 8325 strain staO32 is SAOUHSC_00958 and has amino acid sequence SEQ ID NO: 68 (GI:88194715). Useful staO32 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 68 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 68; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 68, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO32 proteins include variants of SEQ ID NO: 68. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 68. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 68 while retaining at least one epitope of SEQ ID NO: 68. Other fragments omit one or more protein domains. staO33
The 'staO33' antigen is annotated as 'cysteine protease precursor'. In the NCTC 8325 strain staO33 is
SAOUHSC_00987 and has amino acid sequence SEQ ID NO: 69 (GI: 88194744). Useful staO33 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 69 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 69; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 69, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO33 proteins include variants of SEQ ID NO: 69. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 69. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 69 while retaining at least one epitope of SEQ ID NO: 69. The first 29 N- terminal amino acids of SEQ ID NO: 69 can usefully be omitted. Other fragments omit one or more protein domains. staO34
The 'staO34' antigen is annotated as 'glutamyl endopeptidase precursor'. In the NCTC 8325 strain staO34 is SAOUHSC 00988 and has amino acid sequence SEQ ID NO: 70 (GI:88194745). Useful staO34 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 70 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 70; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 70, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO34 proteins include variants of SEQ ID NO: 70. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 70. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 70 while retaining at least one epitope of SEQ ID NO: 70. The first 29 N- terminal amino acids of SEQ ID NO: 70 can usefully be omitted. Other fragments omit one or more protein domains. staO35
The 'staO35' antigen is annotated as 'frnt protein'. In the NCTC 8325 strain staO35 is SAOUHSC_00998 and has amino acid sequence SEQ ID NO: 71 (GI: 88194754).
Useful staO35 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 71 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 71; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 71, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO35 proteins include variants of SEQ ID NO: 71. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 71. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 71 while retaining at least one epitope of SEQ ID NO: 71. The first 25 N- terminal amino acids of SEQ ID NO: 71 can usefully be omitted. Other fragments omit one or more protein domains. staO36
The 'staO36' antigen is annotated as 'iron-regulated protein with leader'. In the NCTC 8325 strain staO36 is SAOUHSC_01084 and has amino acid sequence SEQ ID NO: 72 (GI:88194831).
Useful staO36 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 72 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 72; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 72, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO36 proteins include variants of SEQ ID NO: 72. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 72. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 72 while retaining at least one epitope of SEQ ID NO: 72. The final 27 C- terminal amino acids of SEQ ID NO: 72 can usefully be omitted. The first 32 N-terminal amino acids of SEQ ID NO: 72 can usefully be omitted. Other fragments omit one or more protein domains. staO37
The 'staO37' antigen is annotated as 'iron ABC transporter; iron -binding protein IsdE'. In the NCTC 8325 strain staO37 is SAOUHSC_01085 and has amino acid sequence SEQ ID NO: 73 (GI: 88194832). Useful staO37 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 73 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 73; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 73, wherein V is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO37 proteins include variants of SEQ ID NO: 73. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 73. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 73 while retaining at least one epitope of SEQ ID NO: 73. The first 9 N- terminal amino acids of SEQ ED NO: 73 can usefully be omitted. Other fragments omit one or more protein domains. staO38
The 'staO38' antigen is annotated as 1NPQTN specific sortase B'. hi the NCTC 8325 strain staO38 is
SAOUHSC 01088 and has amino acid sequence SEQ ID NO: 74 (GI:88194835). Useful staO38 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 74 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 74; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 74, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO38 proteins include variants of SEQ ID NO: 74. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 74. Other preferred fragments lack one or more amino acids,(e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 74 while retaining at least one epitope of SEQ ID NO: 74. The first 21 N-terminal amino acids of SEQ ID NO: 74 can usefully be omitted. Other fragments omit one or more protein domains. staO39
The 'staO39' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staO39 is
SAOUHSC Ol 124 and has amino acid sequence SEQ DD NO: 75 (GL88194868).
Useful staO39 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 75 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 75; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 75, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO39 proteins include variants of SEQ ID NO: 75. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 75. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 75 while retaining at least one epitope of SEQ ID NO: 75. The first 22 N-terminal amino acids of SEQ ID NO: 75 can usefully be omitted. Other fragments omit one or more protein domains. sta040 The 'staO4O' antigen is annotated as 'superantigen-like protein1. In the NCTC 8325 strain staO4O is SAOUHSCJ)1125 and has amino acid sequence SEQ ID NO: 76 (GI: 88194869). In the Newman strain it is nwmn_1076 (GI: 151221288).
Useful sta040 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 76 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 76; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 76, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These sta040 proteins include variants of SEQ ID NO: 76. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 76. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 76 while retaining at least one epitope of SEQ ID NO: 76. The first 21 N-terminal amino acids of SEQ ED NO: 76 can usefully be omitted. Other fragments omit one or more protein domains. staO41 The ?sta041' antigen is annotated as 'fibronectin-binding protein A-related'. In the NCTC 8325 strain staO41 is SAOUHSC Ol 175 and has amino acid sequence SEQ ID NO: 77 (GI:88194914).
Useful staO41 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ED NO: 77 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 77; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 77, wherein ?n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO41 proteins include variants of SEQ ID NO: 77. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 77. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 77 while retaining at least one epitope of SEQ ID NO: 77. Other fragments omit one or more protein domains. staO42
The 'staO42' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO42 is
SAOUHSC Ol 180 and has amino acid sequence SEQ ID NO: 78 (GI: 88194919).
Useful staO42 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 78 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 78; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 78, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO42 proteins include variants of SEQ ID NO: 78. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 78. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 78 while retaining at least one epitope of SEQ ID NO: 78. The first 18 N- terminal amino acids of SEQ ID NO: 78 can usefully be omitted. Other fragments omit one or more protein domains. staO43
The 'staO43' antigen is annotated as 'cell wall hydrolase', hi the NCTC 8325 strain staO43 is
SAOUHSC_01219 and has amino acid sequence SEQ ID NO: 79 (GI:88194955).
Useful staO43 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ED NO: 79 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 79; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 79, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These staO43 proteins include variants of SEQ ID NO: 79. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 79. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 79 while retaining at least one epitope of SEQ ID NO: 79. The first 38 N- terminal amino acids of SEQ ID NO: 79 can usefully be omitted. Other fragments omit one or more protein domains. staO44
The 'staO44' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO44 is
SAOUHSC_01508 and has amino acid sequence SEQ ID NO: 80 (GI:88195223).
Useful staO44 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 80 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 80; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 80, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO44 proteins include variants of SEQ ID NO: 80. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 80. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 80 while retaining at least one epitope of SEQ ID NO: 80. The first 17 N- terminal amino acids of SEQ ID NO: 80 can usefully be omitted. Other fragments omit one or more protein domains. staO45
The 'staO45' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO45 is SAOUHSC_01627 and has amino acid sequence SEQ ID NO: 81 (GL88195337).
Useful staO45 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 81 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 81; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 81, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These staO45 proteins include variants of SEQ ID NO: 81. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 81. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ED NO: 81 while retaining at least one epitope of SEQ ID NO: 81. The first 16 N-terminal amino acids of SEQ ID NO: 81 can usefully be omitted. Other fragments omit one or more protein domains. staO46 The 'staO46' antigen is annotated as 'Excalibur protein'. In the NCTC 8325 strain staO46 is SAOUHSC_01918 and has amino acid sequence SEQ ID NO: 82 (GI:88195613).
Useful staO46 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 82 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 82; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 82, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO46 proteins include variants of SEQ ID NO: 82. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 82. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 82 while retaining at least one epitope of SEQ ID NO: 82. The first 53 N-terminal amino acids of SEQ ID NO: 82 can usefully be omitted. Other fragments omit one or more protein domains. staO47
The 'staO47' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO47 is SAOUHSC_01920 and has amino acid sequence SEQ ID NO: 83 (GL88195615).
Useful staO47 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 83 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 83; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 83, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO47 proteins include variants of SEQ ID NO: 83. Preferred fragments of (b) comprise an epitope from SEQ ED NO: 83. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 83 while retaining at least one epitope of SEQ ID NO: 83. The first 18 N-terminal amino acids of SEQ ED NO: 83 can usefully be omitted. Other fragments omit one or more protein domains. staO48
The 'staO48' antigen is annotated as 'intracellular serine protease'. En the NCTC 8325 strain staO48 is SAOUHSC_01949 and has amino acid sequence SEQ ID NO: 84 (GI:88195642).
Useful staO48 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 84 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 84; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 84, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO48 proteins include variants of SEQ ID NO: 84. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 84. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 84 while retaining at least one epitope of SEQ ED NO: 84. The first 27 N- terminal amino acids of SEQ ID NO: 84 can usefully be omitted. Other fragments omit one or more protein domains. staO49
The 'staO49' antigen is annotated as 'protein export protein PrsA'. In the NCTC 8325 strain staO49 is SAOUHSCJ) 1972 and has amino acid sequence SEQ ID NO: 85 (GI: 88195663). In the Newman strain it is nwmn_1733 (GL151221945).
Useful staO49 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 85 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 85; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 85, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO49 proteins include variants of SEQ ID NO: 85. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 85. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 85 while retaining at least one epitope of SEQ ID NO: 85. The first 25 N- terminal amino acids of SEQ ID NO: 85 can usefully be omitted. Other fragments omit one or more protein domains. sta050
The 'staO5O' antigen is annotated as 'staphopain thiol proteinase'. In the NCTC 8325 strain sta050 is SAOUHSC 02127 and has amino acid sequence SEQ ID NO: 86 (GI:88195808).
Useful staO5O antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 86 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 86; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 86, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO5O proteins include variants of SEQ ID NO: 86. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 86. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 86 while retaining at least one epitope of SEQ ID NO: 86. The first 25 N- terminal amino acids of SEQ ID NO: 86 can usefully be omitted. Other fragments omit one or more protein domains. staOSl
The 'staO51' antigen is annotated as 'protein with leader'. In the NCTC 8325 strain staO51 is SAOUHSC 02147 and has amino acid sequence SEQ ID NO: 87 (GI: 88195827).
Useful staO51 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 87 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 87; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 87, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO51 proteins include variants of SEQ ID NO: 87. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 87. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 87 while retaining at least one epitope of SEQ ID NO: 87. The first 24 N- terminal amino acids of SEQ ID NO: 87 can usefully be omitted. Other fragments omit one or more protein domains. staO52
The 'staO52' antigen is annotated as 'ferric hydroxamate receptor 1'. In the NCTC 8325 strain staO52 is SAOUHSC 02246 and has amino acid sequence SEQ ID NO: 88 (GL88195918).
Useful staO52 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 88 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 88; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 88, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO52 proteins include variants of SEQ ID NO: 88. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 88. Other preferred fragments lactone or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 88 while retaining at least one epitope of SEQ ID NO: 88. The first 17 N- terminal amino acids of SEQ ED NO: 88 can usefully be omitted. Other fragments omit one or more protein domains. staO53
The ?sta053' antigen is annotated as 'srdH family protein1. In the NCTC 8325 strain staO53 is SAOUHSC_02257 and has amino acid sequence SEQ ID NO: 89 (GI: 88195928).
Useful staO53 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 89 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 89; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 89, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO53 proteins include variants of SEQ ID NO: 89. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 89. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 89 while retaining at least one epitope of SEQ ID NO: 89. The first 26 N- terminal amino acids of SEQ ID NO: 89 can usefully be omitted. Other fragments omit one or more protein domains. staO54
The 'staO54' antigen is annotated as 'Probable transglycosylase isaA precursor'. In the NCTC 8325 strain staO54 is SAOUHSC 02333 and has amino acid sequence SEQ ID NO: 90 (GI: 88195999). Useful staO54 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 90 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 90; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 90, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO54 proteins include variants of SEQ ID NO: 90. Preferred fragments of (b) comprise an epitope from SEQ ED NO: 90. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 90 while retaining at least one epitope of SEQ ID NO: 90. The first 27 N-terminal amino acids of SEQ ID NO: 90 can usefully be omitted. Other fragments omit one or more protein domains. staO55
The 'staO55' antigen is annotated as 'surface hydrolase'. In the NCTC 8325 strain staO55 is
SAOUHSC_02448 and has amino acid sequence SEQ LD NO: 91 (GL88196100). Useful staO55 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 91 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 91; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 91, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO55 proteins include variants of SEQ ID NO: 91. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 91. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 91 while retaining at least one epitope of SEQ ID NO: 91. The first 31 N- terminal amino acids of SEQ ID NO: 91 can usefully be omitted. Other fragments omit one or more protein domains. staO56
The ?sta056' antigen is annotated as 'hyaluronate lyase'. In the NCTC 8325 strain staO56 is
SAOUHSC_02463 and has amino acid sequence SEQ ID NO: 92 (GI:88196115). Useful staO56 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 92 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 92; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 92, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO56 proteins include variants of SEQ ID NO: 92. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 92. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 92 while retaining at least one epitope of SEQ ID NO: 92. The first 24 N- terminal amino acids of SEQ ID NO: 92 can usefully be omitted. Other fragments omit one or more protein domains. staO57
The 'staO57' antigen is annotated as 'secretory antigen precursor SsaA'. In the NCTC 8325 strain staO57 is SAOUHSC 02576 and has amino acid sequence SEQ ID NO: 93 (GI: 88196220). In the Newman strain it is nwmn_2203 (GI:151222415).
Useful staO57 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 93 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 93; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 93, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These staO57 proteins include variants of SEQ ID NO: 93. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 93. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 93 while retaining at least one epitope of SEQ ID NO: 93. The first 27 N-terminal amino acids of SEQ ID NO: 93 can usefully be omitted. Other fragments omit one or more protein domains. staO58
The 'staO58' antigen is annotated as 'Zn-binding lipoprotein adcA-like1. In the NCTC 8325 strain staO58 is SAOUHSC 02690 and has amino acid sequence SEQ ID NO: 94 (GL88196330).
Useful staO58 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 94 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 94; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 94, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO58 proteins include variants of SEQ ID NO: 94. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 94. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 94 while retaining at least one epitope of SEQ ID NO: 94. The first 20 N- terminal amino acids of SEQ ID NO: 94 can usefully be omitted. Other fragments omit one or more protein domains. staO59
The 'staO59' antigen is annotated as 'gamma-hemolysin h-gamma-ii subunit'. In the NCTC 8325 strain staO59 is SAOUHSC 02708 and has amino acid sequence SEQ ID NO: 95 (GI: 88196348).
Useful staO59 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 95 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 95; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 95, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO59 proteins include variants of SEQ ID NO: 95. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 95. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 95 while retaining at least one epitope of SEQ ID NO: 95. The first 20 N- terminal amino acids of SEQ ID NO: 95 can usefully be omitted. Other fragments omit one or more protein domains. sta060
The 'sta060' antigen is annotated as 'peptide ABC transporter; peptide-binding protein'. In the NCTC
8325 strain sta060 is SAOUHSC_02767 and has amino acid sequence SEQ ID NO: 96
(GI:88196403). Useful sta060 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 96 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 96; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 96, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta060 proteins include variants of SEQ ID NO: 96. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 96. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 96 while retaining at least one epitope of SEQ ID NO: 96. The first 20 N- terminal amino acids of SEQ ID NO: 96 can usefully be omitted. Other fragments omit one or more protein domains. staOόl
The 'staO61' antigen is annotated as 'protein with leader'. In the NCTC 8325 strain staOόl is
SAOUHSC_02783 and has amino acid sequence SEQ ID NO: 97 (GL88196419). Useful staOόl antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 97 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 97; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 97, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO61 proteins include variants of SEQ ID NO: 97. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 97. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 97 while retaining at least one epitope of SEQ ID NO: 97. The first 21 N- terminal amino acids of SEQ ID NO: 97 can usefully be omitted. Other fragments omit one or more protein domains. staO62
The 'staO62' antigen is annotated as 'protein with leader'. In the NCTC 8325 strain staO62 is
SAOUHSC 02788 and has amino acid sequence SEQ ID NO: 98 (GI: 88196424).
Useful staO62 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 98 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 98; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 98, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These staO62 proteins include variants of SEQ ID NO: 98. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 98. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 98 while retaining at least one epitope of SEQ ID NO: 98. The first 22 N- terminal amino acids of SEQ ID NO: 98 can usefully be omitted. Other fragments omit one or more protein domains. staO63
The 'staO63' antigen is annotated as 'aureolysin'. In the NCTC 8325 strain staO63 is
SAOUHSC_02971 and has amino acid sequence SEQ ID NO: 99 (GL88196592).
Useful staO63 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 99 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 99; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 99, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70,
80, 90, 100, 150, 200, 250 or more). These staO63 proteins include variants of SEQ ID NO: 99. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 99. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N- terminus of SEQ ID NO: 99 while retaining at least one epitope of SEQ ID NO: 99. The first 16 N- terminal amino acids of SEQ ED NO: 99 can usefully be omitted. Other fragments omit one or more protein domains. staO64 The 'staO64' antigen is annotated as 'lipase', hi the NCTC 8325 strain staO64 is SAOUHSC 03006 and has amino acid sequence SEQ ID NO: 100 (GI:88196625). hi the Newman strain it is nwmn_2569 (GI: 151222781).
Useful staO64 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 100 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 100; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 100, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO64 proteins include variants of SEQ ID NO: 100. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 100. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 100 while retaining at least one epitope of SEQ ID NO: 100. The first 34 N-terminal amino acids of SEQ ID NO: 100 can usefully be omitted. Other fragments omit one or more protein domains. staO65
The 'staO65' antigen is annotated as '1-phosphatidylinositol phosphodiesterase precursor'. In the NCTC 8325 strain staO65 is SAOUHSC 00051 and has amino acid sequence SEQ ED NO: 101 (GI:88193871).
Useful staO65 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 101 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 101; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ED NO: 101, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO65 proteins include variants of SEQ ED NO: 101. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 101. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 101 while retaining at least one epitope of SEQ ID NO: 101. The first
26 N-terminal amino acids of SEQ ID NO: 101 can usefully be omitted. Other fragments omit one or more protein domains. staO66
The 'staO66' antigen is annotated as 'protein'. In the NCTC 8325 strain staO66 is SAOUHSC 00172 and has amino acid sequence SEQ ID NO: 102 (GI:88193982).
Useful staO66 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 102 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 102; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 102, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO66 proteins include variants of SEQ ID NO: 102. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 102. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 102 while retaining at least one epitope of SEQ ID NO: 102. The first 21 N-terminal amino acids of SEQ ID NO: 102 can usefully be omitted. Other fragments omit one or more protein domains. staO67
The 'staO67' antigen is annotated as 'bacterial extracellular solute-binding protein'. In the NCTC 8325 strain staO67 is SAOUHSC 00176 and has amino acid sequence SEQ ID NO: 103 (GL88193986).
Useful staO67 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 103 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 103; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 103, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO67 proteins include variants of SEQ ID NO: 103. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 103. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 103 while retaining at least one epitope of SEQ ID NO: 103. The first
20 N-terminal amino acids of SEQ ID NO: 103 can usefully be omitted. Other fragments omit one or more protein domains. staO68
The 'staO68' antigen is annotated as 'iron permease FTRl'. In the NCTC 8325 strain staO68 is
SAOUHSC_00327 and has amino acid sequence SEQ ID NO: 104 (GL88194127).
Useful staO68 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 104 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 104; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 104, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO68 proteins include variants of SEQ ID NO: 104. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 104. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-teπninus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 104 while retaining at least one epitope of SEQ ID NO: 104. The final 20 C-terminal amino acids of SEQ ID NO: 104 can usefully be omitted. The first 14 N-terminal amino acids of SEQ ID NO: 104 can usefully be omitted. Other fragments omit one or more protein domains. sta069
The 'staO69' antigen is annotated as 'autolysin precursor'. In the NCTC 8325 strain staO69 is
SAOUHSC_00427 and has amino acid sequence SEQ ID NO: 105 (GL88194219).
Useful staO69 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 105 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 105; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 105, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO69 proteins include variants of SEQ ID NO: 105. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 105. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 105 while retaining at least one epitope of SEQ ID NO: 105. The first
25 N-terminal amino acids of SEQ ID NO: 105 can usefully be omitted. Other fragments omit one or more protein domains. sta070
The 'sta070' antigen is annotated as 'immunogenic secreted precursor-like protein (truncated)'. In the
NCTC 8325 strain sta070 is SAOUHSC 00773 and has amino acid sequence SEQ ID NO: 106
(GL88194535). Useful sta070 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 106 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 106; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 106, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta070 proteins include variants of SEQ ID NO: 106. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 106. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 106 while retaining at least one epitope of SEQ ID NO: 106. The first 24 N-terminal amino acids of SEQ ID NO: 106 can usefully be omitted. Other fragments omit one or more protein domains. staO71
The 'staO71' antigen is annotated as 'hemolysin'. In the NCTC 8325 strain staO71 is SAOUHSC_00854 and has amino acid sequence SEQ ID NO: 107 (GL88194612).
Useful staO71 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 107 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 107; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 107, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO71 proteins include variants of SEQ ID NO: 107. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 107. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 107 while retaining at least one epitope of SEQ ID NO: 107. The first 24 N-terminal amino acids of SEQ ID NO: 107 can usefully be omitted. Other fragments omit one or more protein domains. staO72
The ?sta072' antigen is annotated as 'extramembranal protein'. In the NCTC 8325 strain staO72 is SAOUHSC 00872 and has amino acid sequence SEQ ID NO: 108 (GI: 88194629).
Useful staO72 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 108 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 108; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 108, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO72 proteins include variants of SEQ ID NO: 108. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 108. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 108 while retaining at least one epitope of SEQ ID NO: 108. The first 24 N-terminal amino acids of SEQ ID NO: 108 can usefully be omitted. Other fragments omit one or more protein domains. staO73
The ?sta073' antigen is annotated as 'bifunctional autolysin precursor'. In the NCTC 8325 strain staO73 is SAOUHSC 00994 and has amino acid sequence SEQ ID NO: 109 (GI: 88194750). In the Newman strain it is nwmn_0922 (GI: 151221134). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
Useful staO73 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 109 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ED NO: 109; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 109, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO73 proteins include variants of SEQ ID NO: 109. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 109. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 109 while retaining at least one epitope of SEQ ID NO: 109. The first 24 N-terminal amino acids of SEQ ID NO: 109 can usefully be omitted. Other fragments omit one or more protein domains.
A StaO73 antigen can usefully be included in a composition in combination with a Stal 12 [74].
StaO73 does not adsorb well to aluminium hydroxide adjuvants, so StaO73 present in a composition may be unadsorbed or may be adsorbed to an alternative adjuvant e.g. to an aluminium phosphate. staO74
The 'staO74' antigen is annotated as 'factor essential for methicillin resistance'. In the NCTC 8325 strain staO74 is SAOUHSC 01220 and has amino acid sequence SEQ ID NO: 110 (GL88194956).
Useful staO74 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 110 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 110; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 110, wherein V is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO74 proteins include variants of SEQ ID NO: 110. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 110. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 110 while retaining at least one epitope of SEQ ID NO: 110. Other fragments omit one or more protein domains. staO75
The 'staO75' antigen is annotated as 'insulysin; peptidase family Ml 6'. In the NCTC 8325 strain staO75 is SAOUHSC 01256 and has amino acid sequence SEQ ID NO: 111 (GI: 88194989).
Useful staO75 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 111 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 111; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 111, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO75 proteins include variants of SEQ ID NO: 111. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 111. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 111 while retaining at least one epitope of SEQ ID NO: 111. Other fragments omit one or more protein domains. staO76
The 'staO76' antigen is annotated as 'hydrolase'. In the NCTC 8325 strain staO76 is SAOUHSC 01263 and has amino acid sequence SEQ ID NO: 112 (GI:88194996).
Useful staO76 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 112 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 112; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 112, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO76 proteins include variants of SEQ ID NO: 112. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 112. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 112 while retaining at least one epitope of SEQ ID NO: 112. The first 24 N-terminal amino acids of SEQ ID NO: 112 can usefully be omitted. Other fragments omit one or more protein domains. staO77
The 'staO77' antigen is annotated as 'protein'. In the NCTC 8325 strain staO77 is SAOUHSC 01317 and has amino acid sequence SEQ ID NO: 113 (GI:88195047). Proteomic analysis has revealed that this protein is secreted or surface-exposed.
Useful staO77 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 113 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 113; and/or (b) comprising a fragment, of at least 'n' consecutive amino acids of SEQ ID NO: 113, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO77 proteins include variants of SEQ ID NO: 113. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 113. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 113 while retaining at least one epitope of SEQ ID NO: 113. The first 20 N-terminal amino acids of SEQ ID NO: 113 can usefully be omitted. Other fragments omit one or more protein domains. staO78
The 'staO78' antigen is annotated as 'FtsK/SpoIIIE family protein'. In the NCTC 8325 strain staO78 is SAOUHSC 01857 and has amino acid sequence SEQ ID NO: 114 (GI:88195555).
Useful staO78 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 114 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 114; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 114, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO78 proteins include variants of SEQ ID NO: 114. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 114. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 114 while retaining at least one epitope of SEQ ID NO: 114. Other fragments omit one or more protein domains. staO79 The 'staO79' antigen is annotated as 'serine protease SpIF'. In the NCTC 8325 strain staO79 is SAOUHSC_01935 and has amino acid sequence SEQ ID NO: 115 (GI:88195630).
Useful staO79 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 115 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 115; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 115, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO79 proteins include variants of SEQ ID NO: 115. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 115. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 115 while retaining at least one epitope of SEQ ID NO: 115. The first 36 N-terminal amino acids of SEQ ID NO: 115 can usefully be omitted. Other fragments omit one or more protein domains. sta.080
The 'staO8O' antigen is annotated as 'serine protease SpIE'. In the NCTC 8325 strain staO8O is SAOUHSC 01936 and has amino acid sequence SEQ ID NO: 116 (GI:88195631).
Useful staO8O antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 116 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 116; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 116, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO8O proteins include variants of SEQ ID NO: 116. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 116. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 116 while retaining at least one epitope of SEQ ID NO: 116. The first 36 N-terminal amino acids of SEQ ID NO: 116 can usefully be omitted. Other fragments omit one or more protein domains. staO81
The 'staO81' antigen is annotated as 'serine protease SpID (EC:3.4.21.19)'. In the NCTC 8325 strain staO81 is SAOUHSC 01938 and has amino acid sequence SEQ ID NO: 170 (GI:88195633).
Useful staO81 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 170 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 170; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 170, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO81 proteins include variants of SEQ ID NO: 170. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 170. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 170 while retaining at least one epitope of SEQ ID NO: 170. The first 36 N-terminal amino acids of SEQ ID NO: 170 can usefully be omitted. Other fragments omit one or more protein domains. staO82
The 'staO82' antigen is annotated as 'serine protease SpIC. In the NCTC 8325 strain staO82 is SAOUHSC 01939 and has amino acid sequence SEQ ID NO: 117 (GI: 88195634). Useful staO82 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 117 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 117; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 117, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO82 proteins include variants of SEQ ID NO:
117. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 117. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 117 while retaining at least one epitope of SEQ ID NO: 117. The first 36 N-terminal amino acids of SEQ ID NO: 117 can usefully be omitted. Other fragments omit one or more protein domains. staO83
The 'staO83' antigen is annotated as 'serine protease SpIB'. In the NCTC 8325 strain staO83 is SAOUHSC 01941 and has amino acid sequence SEQ ID NO: 118 (GI:88195635).
Useful staO83 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 118 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 118; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 118, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO83 proteins include variants of SEQ ID NO:
118. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 118. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 118 while retaining at least one epitope of SEQ ID NO: 118. The first 36 N-terminal amino acids of SEQ ID NO: 118 can usefully be omitted. Other fragments omit one or more protein domains. staO84
The 'staO84' antigen is annotated as 'serine protease SpIA'. In the NCTC 8325 strain staO84 is SAOUHSC_01942 and has amino acid sequence SEQ ID NO: 119 (GI:88195636).
Useful staO84 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 119 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 119; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 119, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO84 proteins include variants of SEQ ID NO:
119. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 119. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 119 while retaining at least one epitope of SEQ ID NO: 119. The first 35 N-terminal amino acids of SEQ ID NO: 119 can usefully be omitted. Other fragments omit one or more protein domains. staO85
The ?sta085' antigen is annotated as 'staphylokinase precursor'. In the NCTC 8325 strain staO85 is
SAOUHSC_02171 and has amino acid sequence SEQ ID NO: 120 (GI:88195848).
Useful staO85 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 120 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 120; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 120, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150 or more). These staO85 proteins include variants of SEQ ID NO: 120. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 120. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
N-terminus of SEQ ID NO: 120 while retaining at least one epitope of SEQ ID NO: 120. The first 27
N-terminal amino acids of SEQ ID NO: 120 can usefully be omitted. Other fragments omit one or more protein domains. staO86
The ?sta086' antigen is annotated as 'OxaA-like protein'. In the NCTC 8325 strain staO86 is
SAOUHSC 02327 and has amino acid sequence SEQ ID NO: 121 (GI:88195993).
Useful staO86 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 121 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 121; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 121, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO86 proteins include variants of SEQ ID NO: 121. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 121. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 121 while retaining at least one epitope of SEQ ID NO: 121. The first
19 N-terminal amino acids of SEQ ID NO: 121 can usefully be omitted. Other fragments omit one or more protein domains. staO87
The 'staO87' antigen is annotated as 'teicoplanin resistance protein TcaA'. In the NCTC 8325 strain staO87 is SAOUHSC 02635 and has amino acid sequence SEQ ID NO: 122 (GI: 88196276).
Useful staO87 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 122 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 122; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 122, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO87 proteins include variants of SEQ ID NO: 122. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 122. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 122 while retaining at least one epitope of SEQ ID NO: 122. Other fragments omit one or more protein domains. staO88
The 'staO88' antigen is annotated as 'esterase', hi the NCTC 8325 strain staO88 is SAOUHSC 02844 and has amino acid sequence SEQ ID NO: 123 (GI:88196477).
Useful staO88 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 123 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 123; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 123, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO88 proteins include variants of SEQ ID NO: 123. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 123. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 123 while retaining at least one epitope of SEQ ID NO: 123. The first 18 N-terminal amino acids of SEQ ID NO: 123 can usefully be omitted. Other fragments omit one or more protein domains. staO89
The 'staO89' antigen is annotated as 'LysM domain protein'. In the NCTC 8325 strain staO89 is SAOUHSC 02855 and has amino acid sequence SEQ ID NO: 124 (GI:88196486).
Useful staO89 antigens can elicit an antibody (e.g. when administered to a human) that recognises
SEQ ID NO: 124 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 124; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 124, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These staO89 proteins include variants of SEQ ID NO: 124. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 124. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 124 while retaining at least one epitope of SEQ ID NO: 124. The first 20 N-terminal amino acids of SEQ ID NO: 124 can usefully be omitted. Other fragments omit one or more protein domains. sta090 The 'sta090' antigen is annotated as 'LysM domain protein'. In the NCTC 8325 strain sta090 is SAOUHSC_02883 and has amino acid sequence SEQ ID NO: 125 (GI:88196512).
Useful sta090 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 125 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 125; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 125, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These sta090 proteins include variants of SEQ ID NO: 125. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 125. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 125 while retaining at least one epitope of SEQ ID NO: 125. The first 26 N-terminal amino acids of SEQ ID NO: 125 can usefully be omitted. Other fragments omit one or more protein domains. staO91 The 'staO91' antigen is annotated as 'lipoprotein'. In the NCTC 8325 strain staO91 is SAOUHSC 00685 and has amino acid sequence SEQ ID NO: 126 (GI:88194450).
Useful staO91 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 126 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 126; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 126, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These staO91 proteins include variants of SEQ ID NO: 126. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 126. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 126 while retaining at least one epitope of SEQ ID NO: 126. The first 15 N-terminal amino acids of SEQ ID NO: 126 can usefully be omitted. Other fragments omit one or more protein domains. staO92
The 'staO92' antigen is annotated as 'M23/M37 peptidase domain protein'. In the NCTC 8325 strain staO92 is SAOUHSC 00174 and has amino acid sequence SEQ ID NO: 127 (GI:88193984).
Useful staO92 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 127 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 127; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 127, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These staO92 proteins include variants of SEQ ID NO: 127. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 127. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 127 while retaining at least one epitope of SEQ ID NO: 127. The first 25 N-terminal amino acids of SEQ ID NO: 127 can usefully be omitted. Other fragments omit one or more protein domains. staO93
The 'staO93' antigen is annotated as 'protein'. In the NCTC 8325 strain staO93 is SAOUHSC O 1854 and has amino acid sequence SEQ ID NO: 128 (GI: 88195552).
Useful staO93 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 128 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 128; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ED NO: 128, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO93 proteins include variants of SEQ ID NO: 128. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 128. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 128 while retaining at least one epitope of SEQ ED NO: 128. Other fragments omit one or more protein domains. staO94
The 'staO94' antigen is annotated as 'protein'. In the NCTC 8325 strain staO94 is SAOUHSC Ol 512 and has amino acid sequence SEQ ID NO: 129 (GI: 88195226). Useful staO94 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 129 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 129; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 129, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO94 proteins include variants of SEQ ID NO: 129. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 129. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 129 while retaining at least one epitope of SEQ ID NO: 129. The first 17 N-terminal amino acids of SEQ ID NO: 129 can usefully be omitted. Other fragments omit one or more protein domains. staO95
The 'staO95' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staO95 is SAOUHSC 00383 and has amino acid sequence SEQ ID NO: 130 (GI:88194180). In the Newman strain it is nwmn_0388 (GI: 151220600). Useful staO95 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 130 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 130; and/or (b) comprising a fragment of at least W consecutive amino acids of SEQ ID NO: 130, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO95 proteins include variants of SEQ ID NO:
130. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 130. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 130 while retaining at least one epitope of SEQ ID NO: 130. The first 32 N-terminal amino acids of SEQ ID NO: 130 can usefully be omitted. Other fragments omit one or more protein domains. staO96
The 'staO96' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staO96 is
SAOUHSC 00384 and has amino acid sequence SEQ ID NO: 131 (GI:88194181). Useful staO96 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 131 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 131; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 131, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO96 proteins include variants of SEQ ID NO:
131. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 131. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 131 while retaining at least one epitope of SEQ ID NO: 131. The first 30 N-terminal amino acids of SEQ ID NO: 131 can usefully be omitted. Other fragments omit one or more protein domains. staO97
The 'staO97' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staO97 is SAOUHSC_00386 and has amino acid sequence SEQ ID NO: 132 (GI:88194182).
Useful staO97 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 132 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 132; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 132, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO97 proteins include variants of SEQ ID NO: 132. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 132. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 132 while retaining at least one epitope of SEQ ID NO: 132. The first 30 N-terminal amino acids of SEQ ID NO: 132 can usefully be omitted. Other fragments omit one or more protein domains. staO98
The 'staO98' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain staO98 is SAOUHSC_00389 and has amino acid sequence SEQ ID NO: 133 (GL88194184). In the Newman strain it is nwmn_0391 (GI: 151220603).
Useful staO98 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 133 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 133; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 133, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These staO98 proteins include variants of SEQ ID NO: 133. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 133. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 133 while retaining at least one epitope of SEQ ID NO: 133. The first
30 N-terminal amino acids of SEQ ID NO: 133 can usefully be omitted. Other fragments omit one or more protein domains. staO99
The ?sta099' antigen is annotated as 'superantigen-like protein 5'. In the NCTC 8325 strain staO99 is SAOUHSC_00390 and has amino acid sequence SEQ ID NO: 134 (GL88194185).
Useful staO99 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 134 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 134; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 134, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These staO99 proteins include variants of SEQ ID NO: 134. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 134. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 134 while retaining at least one epitope of SEQ ID NO: 134. The first 30 N-terminal amino acids of SEQ ID NO: 134 can usefully be omitted. Other fragments omit one or more protein domains. stalOO
The 'stalOO' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain stalOO is
SAOUHSC_00391 and has amino acid sequence SEQ ID NO: 135 (GI:88194186).
Useful stalOO antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 135 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 135; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 135, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalOO proteins include variants of SEQ ID NO: 135. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 135. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 135 while retaining at least one epitope of SEQ ID NO: 135. The first
30 N-terminal amino acids of SEQ ID NO: 135 can usefully be omitted. Other fragments omit one or more protein domains. stalOl
The 'stalOl' antigen is annotated as 'superantigen-like protein 7'. In the NCTC 8325 strain stalOl is SAOUHSC_00392 and has amino acid sequence SEQ ID NO: 136 (GL88194187). hi the Newman strain it is nwmn_0394 (GI: 151220606). Useful stalOl antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 136 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 136; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 136, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalOl proteins include variants of SEQ ID NO: 136. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 136. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 136 while retaining at least one epitope of SEQ ID NO: 136. The first 30 N-terminal amino acids of SEQ ID NO: 136 can usefully be omitted. Other fragments omit one or more protein domains. stalO2
The 'stalO2' antigen is annotated as 'superantigen-like protein', hi the NCTC 8325 strain stalO2 is
SAOUHSC_00393 and has amino acid sequence SEQ ID NO: 137 (GI:88194188).
Useful stalO2 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 137 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 137; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 137, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalO2 proteins include variants of SEQ ID NO: 137. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 137. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 137 while retaining at least one epitope of SEQ ID NO: 137. The first
17 N-terminal amino acids of SEQ ID NO: 137 can usefully be omitted. Other fragments omit one or more protein domains. stalO3
The 'stalO3' antigen is annotated as 'superantigen-like protein', hi the NCTC 8325 strain stalO3 is
SAOUHSC_00394 and has amino acid sequence SEQ ID NO: 138 (GI:88194189).
Useful stalO3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 138 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 138; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 138, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalO3 proteins include variants of SEQ ID NO: 138. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 138. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 138 while retaining at least one epitope of SEQ ID NO: 138. The first 23 N-terminal amino acids of SEQ ID NO: 138 can usefully be omitted. Other fragments omit one or more protein domains. stalO4 The 'stalO4' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain stalO4 is SAOUHSC 00395 and has amino acid sequence SEQ ID NO: 139 (GI:88194190).
Useful stalO4 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 139 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 139; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 139, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalO4 proteins include variants of SEQ ID NO:
139. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 139. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 139 while retaining at least one epitope of SEQ ID NO: 139. Other fragments omit one or more protein domains. stalO5
The 'stalO5' antigen is annotated as 'superantigen-like protein'. In the NCTC 8325 strain stalO5 is SAOUHSC 00399 and has amino acid sequence SEQ ID NO: 140 (GI:88194194). In the Newman strain it is nwmn_0400 (GI: 151220612).
Useful stalO5 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 140 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 140; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 140, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalO5 proteins include variants of SEQ ID NO:
140. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 140. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 140 while retaining at least one epitope of SEQ ID NO: 140. The first 30 N-terminal amino acids of SEQ ID NO: 140 can usefully be omitted. Other fragments omit one or more protein domains. stalOό The 'stalOό' antigen is annotated as 'hypothetical protein'. In the NCTC 8325 strain stalOό is SAOUHSC Ol 115 and has amino acid sequence SEQ ID NO: 141 (GI: 88194861). Useful stalOό antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 141 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 141; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 141, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These stalOό proteins include variants of SEQ ID NO: 141. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 141. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 141 while retaining at least one epitope of SEQ ID NO: 141. The first 16 N-terminal amino acids of SEQ ID NO: 141 can usefully be omitted. Other fragments omit one or more protein domains. stalO7
The 'stalO7' antigen is annotated as 'hypothetical protein1. In the NCTC 8325 strain stalO7 is SAOUHSC_00354 and has amino acid sequence SEQ ID NO: 177 (GI:88194153).
Useful stalO7 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 177 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 177; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 177, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stalO7 proteins include variants of SEQ ID NO: 177. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 177. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 177 while retaining at least one epitope of SEQ ID NO: 177. The first 35 N-terminal amino acids of SEQ ID NO: 177 can usefully be omitted. Other fragments omit one or more protein domains. stalO8
The 'stalO8' antigen is annotated as 'hypothetical protein', hi the NCTC 8325 strain stalO8 is SAOUHSC_00717 and has amino acid sequence SEQ ID NO: 178 (GI:88194482).
Useful stalO8 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 178 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 178; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 178, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These stalO8 proteins include variants of SEQ ID NO: 178. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 178. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 178 while retaining at least one epitope of SEQ ID NO: 178. The first 20 N-terminal amino acids of SEQ ID NO: 178 can usefully be omitted. Other fragments omit one or more protein domains. stalO9
The ?stal09' antigen is annotated as "N-acetylmuramoyl-L-alanine amidase '. In the NCTC 8325 strain stalO9 is SAOUHSC_02979 and has amino acid sequence SEQ ID NO: 179 (GI: 88196599).
Useful stalO9 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 179 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 179; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 179, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These stalO9 proteins include variants of SEQ ID NO: 179. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 179. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 179 while retaining at least one epitope of SEQ ID NO: 179. The first
27 N-terminal amino acids of SEQ ID NO: 179 can usefully be omitted. Other fragments omit one or more protein domains. stallO
The 'stal lO' antigen is annotated as 'hypothetical protein'. In the NCTC 8325 strain stal lO is
SAOUHSC_01039 and has amino acid sequence SEQ ID NO: 180 (GI:88194791).
Useful stal lO antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 180 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 180; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 180, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These stal 10 proteins include variants of SEQ ID NO: 180. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 180. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 180 while retaining at least one epitope of SEQ ID NO: 180. The first
19 N-terminal amino acids of SEQ ID NO: 180 can usefully be omitted. Other fragments omit one or more protein domains. stalll
The 'stal l I' antigen is annotated as 'hypothetical protein', hi the NCTC 8325 strain stal l l is SAOUHSC 01005 and has amino acid sequence SEQ ID NO: 181 (GI: 88194760).
Useful stal l l antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 181 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 181; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 181, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These stal l l proteins include variants of SEQ ID NO: 181. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 181. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 181 while retaining at least one epitope of SEQ ID NO: 181. The first 20 N-terminal amino acids of SEQ ID NO: 181 can usefully be omitted. Other fragments omit one or more protein domains. stain
The 'stal l2' antigen is annotated as a putative 'ABC transporter, substrate-binding protein'. In the NCTC 8325 strain stal l2 is SAOUHSC 00634 and has amino acid sequence SEQ ID NO: 182 (GI: 88194402). Useful stal l2 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 182 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 182; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 182, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These stal 12 proteins include variants of SEQ ID NO: 182. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 182. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 182 while retaining at least one epitope of SEQ ID NO: 182. The first 17 N-terminal amino acids of SEQ ID NO: 182 can usefully be omitted. Other fragments omit one or more protein domains.
A Stal 12 antigen can usefully be included in a composition in combination with a StaO73 [74]. stalB
The 'stal 13' antigen is annotated as 'hypothetical protein'. In the NCTC 8325 strain stal 13 is SAOUHSC_00728 and has amino acid sequence SEQ ID NO: 183 (GL88194493). Useful stal l3 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 183 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 183; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 183, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These stal 13 proteins include variants of SEQ ID NO: 183. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 183. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 183 while retaining at least one epitope of SEQ ID NO: 183. The first 173 N-terminal amino acids of SEQ ID NO: 183 can usefully be omitted. Other fragments omit one or more protein domains. stalH
The 'stal 14' antigen is annotated as 'hypothetical protein'. In the NCTC 8325 strain stal 14 is SAOUHSC_00810 and has amino acid sequence SEQ ID NO: 184 (GL88194570).
Useful stal 14 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 184 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 184; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 184, wherein V is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These stal 14 proteins include variants of SEQ ID NO: 184. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 184. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 184 while retaining at least one epitope of SEQ ID NO: 184. Other fragments omit one or more protein domains. stalls
The 'stal 15' antigen is annotated as 'hypothetical protein'. In the NCTC 8325 strain stal 15 is
SAOUHSC_00817 and has amino acid sequence SEQ ID NO: 185 (GI:88194576). Useful stal 15 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 185 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 185; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 185, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These stall5 proteins include variants of SEQ ID NO: 185. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 185. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 185 while retaining at least one epitope of SEQ ID NO: 185. The first 18 N-terminal amino acids of SEQ ID NO: 185 can usefully be omitted. Other fragments omit one or more protein domains. stallό
The 'stal 16' antigen is annotated as 'formyl peptide receptor-like 1 inhibitory protein '. In the NCTC 8325 strain stal lό is SAOUHSCJ)1112 and has amino acid sequence SEQ ID NO: 186 (GI:88194858).
Useful stal lό antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 186 and/or may comprise an amino acid sequence: (a) having 50% or more identity
(e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 186; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 186, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100 or more). These stal lό proteins include variants of SEQ ID NO: 186. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 186. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
N-terminus of SEQ ID NO: 186 while retaining at least one epitope of SEQ ID NO: 186. The first 20
N-terminal amino acids of SEQ ID NO: 186 can usefully be omitted. Other fragments omit one or more protein domains. stall7
The 'stal 17' antigen is annotated as 'truncated beta-hemolysin'. In the NCTC 8325 strain stal 17 is
SAOUHSC_02240 and has amino acid sequence SEQ ID NO: 187 (GI:88195913).
Useful stal 17 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 187 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 187; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 187, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These stal 17 proteins include variants of SEQ ID NO: 187. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 187. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 187 while retaining at least one epitope of SEQ ID NO: 187. Other fragments omit one or more protein domains. stall8
The 'stal lδ' antigen is annotated as 'cell division protein FtsZ'. In the NCTC 8325 strain stal lδ is SAOUHSCJ)1150 and has amino acid sequence SEQ ID NO: 188 (GI: 88194892).
Useful stal lδ antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 188 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 188; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 188, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These stal 18 proteins include variants of SEQ ID NO: 188. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 188. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 188 while retaining at least one epitope of SEQ ID NO: 188. Other fragments omit one or more protein domains. stal 19
The 'stal 19' antigen is annotated as 'thioredoxin'. In the NCTC 8325 strain stal 19 is SAOUHSCJ)1100 and has amino acid sequence SEQ ID NO: 200 (GI: 88194846).
Useful stal 19 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 200 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 200; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 200, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These stal 19 proteins include variants of SEQ ID NO: 200. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 200. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 200 while retaining at least one epitope of SEQ ID NO: 200. Other fragments omit one or more protein domains. stal20 The 'stal 20' antigen is annotated as ' alkyl hydroperoxide reductase subunit C. In the NCTC 8325 strain stal20 is SAOUHSC 00365 and has amino acid sequence SEQ ID NO: 201 (GI:88194163).
Useful stal 20 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 201 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 201; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 201, wherein V is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These stal20 proteins include variants of SEQ ID NO: 201. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 201. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 201 while retaining at least one epitope of SEQ ID NO: 201. Other fragments omit one or more protein domains.
NW_6
The 1NW o' antigen is annotated as 'secreted von Willebrand factor-binding protein precursor'. In the Newman strain NW 6 is NWMN 0757 and has amino acid sequence SEQ ID NO: 142 (GI: 151220969).
Useful NW 6 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 142 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 142; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 142, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These NW 6 proteins include variants of SEQ ID NO: 142. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 142. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 142 while retaining at least one epitope of SEQ ID NO: 142. The first 13 N-terminal amino acids of SEQ ID NO: 142 can usefully be omitted. Other fragments omit one or more protein domains.
NW_9
The "NW 9' antigen is annotated as 'lipoprotein'. In the Newman strain NW 9 is NWMN 0958 and has amino acid sequence SEQ ID NO: 143 (GI: 151221170).
Useful NW 9 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 143 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 143; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 143, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200 or more). These NW 9 proteins include variants of SEQ ID NO: 143. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 143. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 143 while retaining at least one epitope of SEQ ID NO: 143. The first 19 N-terminal amino acids of SEQ ID NO: 143 can usefully be omitted. Other fragments omit one or more protein domains. NWJO
The 1NW lO' antigen is annotated as 'fibrinogen binding-related protein'. In the Newman strain
NW_10 is NWMN_1066 and has amino acid sequence SEQ ID NO: 144 (GL151221278).
Useful NW lO antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 144 and/or may comprise an amino acid sequence: (a) having 50% or more identity {e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 144; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 144, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These NW lO proteins include variants of SEQ ID NO: 144. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 144. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 144 while retaining at least one epitope of SEQ ID NO: 144. The first 20 N-terminal amino acids of SEQ ID NO: 144 can usefully be omitted. Other fragments omit one or more protein domains.
NWJ
The rNW_7' antigen is annotated as 'staphylococcal complement inhibitor SCIN'. hi the Newman strain NW 7 is NWMN 1876 and has amino acid sequence SEQ ID NO: 145 (GI: 151222088).
Useful NW 7 antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 145 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,
99.5% or more) to SEQ ID NO: 145; and/or (b) comprising a fragment of at least V consecutive amino acids of SEQ ID NO: 145, wherein 'n' is 7 or more {e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,
40, 50, 60, 70, 80, 90, 100 or more). These NW 7 proteins include variants of SEQ ID NO: 145. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 145. Other preferred fragments lack one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the
N-terminus of SEQ ID NO: 145 while retaining at least one epitope of SEQ ID NO: 145. The first 17
N-terminal amino acids of SEQ ID NO: 145 can usefully be omitted. Other fragments omit one or more protein domains.
NW_8
The 1NWJ' antigen is annotated as 'chemotaxis-inhibiting protein CHIPS'. In the Newman strain
NWJ is NWMN_1877 and has amino acid sequence SEQ ID NO: 146 (GI: 151222089).
Useful NWJ antigens can elicit an antibody {e.g. when administered to a human) that recognises SEQ ID NO: 146 and/or may comprise an amino acid sequence: (a) having 50% or more identity
{e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 146; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 146, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more). These NWJ proteins include variants of SEQ ID NO: 146. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 146. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 146 while retaining at least one epitope of SEQ ID NO: 146. The first 19 N-terminal amino acids of SEQ ID NO: 146 can usefully be omitted. Other fragments omit one or more protein domains. NWJ
The 1NWJ' antigen is annotated as 'enterotoxin type A precursor'. In the Newman strain NWJ is NWMNJ 883 and has amino acid sequence SEQ ID NO: 147 (GI: 151222095).
Useful NWJ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 147 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 147; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 147, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These NWJ proteins include variants of SEQ ID NO: 147. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 147. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 147 while retaining at least one epitope of SEQ ID NO: 147. The first 16 N-terminal amino acids of SEQ ID NO: 147 can usefully be omitted. Other fragments omit one or more protein domains. NWJ
The TsTWJ' antigen is annotated as 'lipoprotein'. In the Newman strain NWJ is NWMN 1924 and has amino acid sequence SEQ ID NO: 148 (GI: 151222136).
Useful NWJ antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 148 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 148; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 148, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150 or more). These NWJ proteins include variants of SEQ ID NO: 148. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 148. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C- terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 148 while retaining at least one epitope of SEQ ID NO: 148. The first 17 N-terminal amino acids of SEQ ID NO: 148 can usefully be omitted. Other fragments omit one or more protein domains.
NWJ
The 'NW_5' antigen is annotated as 'cell wall surface anchor family protein'. In the Newman strain NWJ is NWMN 2392 and has amino acid sequence SEQ ID NO: 149 (GI: 151222604).
Useful NW 5 antigens can elicit an antibody (e.g. when administered to a human) that recognises SEQ ID NO: 149 and/or may comprise an amino acid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 149; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 149, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These NWJ proteins include variants of SEQ ID NO: 149. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 149. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 149 while retaining at least one epitope of SEQ ID NO: 149. The first 52 N-terminal amino acids of SEQ ID NO: 149 can usefully be omitted. Other fragments omit one or more protein domains.
Hybrid polypeptides
Antigens used in the invention may be present in the composition as individual separate polypeptides. Where more than one antigen is used, however, they do not have to be present as separate polypeptides. Instead, at least two (e.g. 2, 3, 4, 5, or more) antigens can be expressed as a single polypeptide chain (a 'hybrid' polypeptide). Hybrid polypeptides offer two main advantages: first, a polypeptide that may be unstable or poorly expressed on its own can be assisted by adding a suitable hybrid partner that overcomes the problem; second, commercial manufacture is simplified as only one expression and purification need be employed in order to produce two polypeptides which are both antigenically useful.
The hybrid polypeptide may comprise two or more polypeptide sequences from the first antigen group. The hybrid polypeptide may comprise one or more polypeptide sequences from the first antigen group and one or more polypeptide sequences from the second antigen group. Moreover, the hybrid polypeptide may comprise two or more polypeptide sequences from each of the antigens listed above, or two or more variants of the same antigen in the cases in which the sequence has partial variability across strains.
Hybrids consisting of amino acid sequences from two, three, four, five, six, seven, eight, nine, or ten antigens are useful, hi particular, hybrids consisting of amino acid sequences from two, three, four, or five antigens are preferred, such as two or three antigens. Different hybrid polypeptides may be mixed together in a single formulation. Hybrids may be combined with non-hybrid antigens selected from the first, second or third antigen groups. Within such combinations, an antigen may be present in more than one hybrid polypeptide and/or as a non- hybrid polypeptide. It is preferred, however, that an antigen is present either as a hybrid or as a non-hybrid, but not as both.
The hybrid polypeptides can also be combined with conjugates or non-S.aureus antigens as described above.
Hybrid polypeptides can be represented by the formula NH2-A- (-X-L-Jn-B-COOH, wherein: X is an amino acid sequence of a S.aureus antigen, as described above; L is an optional linker amino acid sequence; A is an optional N-terminal amino acid sequence; B is an optional C-terminal amino acid sequence; n is an integer of 2 or more {e.g. 2, 3, 4, 5, 6, etc.). Usually n is 2 or 3.
If a -X- moiety has a leader peptide sequence in its wild-type form, this may be included or omitted in the hybrid protein. In some embodiments, the leader peptides will be deleted except for that of the -X- moiety located at the N-terminus of the hybrid protein i.e. the leader peptide of Xi will be retained, but the leader peptides of X2 ... Xn will be omitted. This is equivalent to deleting all leader peptides and using the leader peptide of Xi as moiety -A-.
For each n instances of {-X-L-}, linker amino acid sequence -L- may be present or absent. For instance, when «=2 the hybrid may be NH2-X1-Li-X2-L2-COOH, NH2-X1-X2-COOH, NH2-Xi-L1-X2- COOH, NH2-X1-X2-L2-COOH, etc. Linker amino acid sequence(s) -L- will typically be short {e.g. 20 or fewer amino acids i.e. 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples comprise short peptide sequences which facilitate cloning, poly-glycine linkers {i.e. comprising GIyn where n = 2, 3, 4, 5, 6, 7, 8, 9, 10 or more), and histidine tags {i.e. Hisn where n = 3, 4, 5, 6, 7, 8, 9, 10 or more). Other suitable linker amino acid sequences will be apparent to those skilled in the art. A useful linker is GSGGGG (SEQ ID NO: 171) or GSGSGGGG (SEQ ID NO: 172), with the Gly-Ser dipeptide being formed from a BamΗl restriction site (or two of them, to form the SEQ ID NO: 230 tetrapeptide), thus aiding cloning and manipulation, and the (GIy)4 tetrapeptide (SEQ ID NO: 227) being a typical poly-glycine linker. Other suitable linkers, particularly for use as the final Ln are ASGGGS (SEQ ID NO: 173 e.g. encoded by SEQ ID NO: 174) or a Leu-Glu dipeptide.
-A- is an optional N-terminal amino acid sequence. This will typically be short {e.g. 40 or fewer amino acids i.e. 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19,
18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples include leader sequences to direct protein trafficking, or short peptide sequences which facilitate cloning or purification {e.g. histidine tags i.e. HiSn where n = 3, 4, 5, 6, 7, 8, 9, 10 or more). Other suitable N-terminal amino acid sequences will be apparent to those skilled in the art. IfX] lacks its own N-terminus methionine, -A- is preferably an oligopeptide {e.g. with 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) which provides a
N-terminus methionine e.g. Met-Ala-Ser, or a single Met residue. -B- is an optional C-terminal amino acid sequence. This will typically be short (e.g. 40 or fewer amino acids i.e. 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples include sequences to direct protein trafficking, short peptide sequences which facilitate cloning or purification (e.g. comprising histidine tags i.e. Hisπ where n = 3, 4, 5, 6, 7, 8, 9, 10 or more, such as SEQ ID NO: 226), or sequences which enhance protein stability. Other suitable C-terminal amino acid sequences will be apparent to those skilled in the art.
One hybrid polypeptide of the invention may include both EsxA and EsxB antigens. These may be in either order, N- to C- terminus. SEQ ID NOs: 151 ('EsxAB'; encoded by SEQ ID NO: 169) and 152 ('EsxBA') are examples of such hybrids, both having hexapeptide linkers ASGGGS (SEQ ID NO: 173). Another 'EsxAB' hybrid comprises SEQ ID NO: 241, which may be provided with a N-terminus methionine (e.g. SEQ ID NO: 250).
Another hybrid polypeptide of the invention may include both SdrD and SdrE antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 168 ('SdrED') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include both CIfB and SdrD antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 202 ('ClfB-SdrD') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 203 ('SdrD-ClfB') is another example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 211 ('ClfB-N3-sdrD-N3') is another example of such a hybrid, where the N3 fragments of CIfB and SdrD are joined by hexapeptide linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include both IsdA and EsxA antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 204 ('IsdA-EsxA') is an example of such a hybrid, having a hexapeptide linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 209 ('isdA40-184-esxA') is another example of such a hybrid, in which ISdA4O-I84 is joined to EsxA via linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include both IsdA and sta006 antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 221 ('isdA40-184-sta006') is an example of such a hybrid, in which ISdA40-I84 is joined to Sta006 via hexapeptide linker ASGGGS (SEQ ID NO: 173). Another hybrid polypeptide of the invention may include both HIa and sta006 antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 222 (ΗlaH35L-staOO6') is an example of such a hybrid, in which a H35L mutant of HIa is joined to Sta006 via hexapeptide linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include both HIa and Emp antigens. These may be in either order, N- to C- terminus. SEQ ID NO: 205 ('HlaH35L-Emp') is an example of such a hybrid, in which a H35L mutant HIa is joined to Emp via linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 206 ('Hla27-76-Emp') is another example of such a hybrid, in which a HIa fragment is joined to Emp via linker ASGGGS (SEQ ID NO: 173); SEQ ID NO: 207 is a H35L mutant of SEQ ID NO: 206. SEQ ID NO: 208 ('HlaPSGS-Emp') is another example of such a hybrid, in which a HIa mutant is joined to Emp via linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include IsdA and EsxA and EsxB antigens. These may be in any order, N- to C- terminus. SEQ ID NO: 210 ('isdA40-184-esxAB') is an example of such a triple hybrid, in which ISdA40-I84 is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173). The EsxAB already includes the same linker, so SEQ ID NO: 210 includes two of these linkers. SEQ ID NO: 212 ('IsdA-esxAB') is another example of such a triple hybrid, in which IsdA is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).
Another hybrid polypeptide of the invention may include HIa and EsxA and EsxB antigens. These may be in any order, N- to C- terminus. SEQ ID NO: 220 ('HlaH35L-esxAB') is an example of such a triple hybrid, in which a H35L mutant of HIa is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173). The EsxAB already includes the same linker, so SEQ ID NO: 220 includes two of these linkers. Another example of a hybrid polypeptide including HIa and EsxA and EsxB antigens is SEQ ID NO: 237 ('HlaH35L-esxAB' as used in the examples), in which a H35L mutant of HIa is joined to EsxA via linker APTARG (SEQ ID NO: 239) to replace its N-terminus, then to EsxB via linker ASGGGS (SEQ ID NO: 173) to replace its N-terminus. This hybrid can be provided with a suitable N-terminal sequence such as SEQ ID NO: 240.
Another hybrid polypeptide of the invention may include sta006 and EsxA and EsxB antigens. These may be in any order, N- to C- terminus. SEQ ID NO: 223 ('sta006-esxAB') is an example of such a triple hybrid, in which sta006 is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173). The EsxAB already includes the same linker, so SEQ ID NO: 223 includes two of these linkers. Another example of a hybrid polypeptide including sta006 and EsxA and EsxB antigens is SEQ ID NO: 238 ('staOOό- esxAB' as used in the examples), in which a staOOό is joined to EsxA via linker APTARG (SEQ ID NO: 239) to replace its N-terminus, then to EsxB via linker ASGGGS (SEQ ID NO: 173) to replace its N-terminus. This hybrid can be provided with a suitable N-terminal sequence such as SEQ ID NO: 240. Usefully, these hybrid polypeptides can elicit an antibody (e.g. when administered to a human) that recognise each of the wild-type staphylococcal proteins (e.g. as shown in the sequence listing) represented in the hybrid e.g. which recognise both wild-type EsxA and wild-type EsxB, or which recognise both wild-type SdrD and wild-type SdrE, or which recognise both wild-type SdrD and wild-type CIfB, or which recognise both wild-type IsdA and wild-type EsxA, or which recognise both wild-type IsdA and wild-type staOOό, or which recognise both wild-type HIa and wild-type staOOό, or which recognise both wild-type HIa and wild-type Emp, or which recognise wild-type IsdA and wild-type EsxA and wild-type EsxB, or which recognise wild-type HIa and wild-type EsxA and wild-type EsxB, or which recognise wild-type sta006 and wild-type EsxA and wild-type EsxB.
Polypeptides used with the invention
Polypeptides used with the invention can take various forms (e.g. native, fusions, glycosylated, non-glycosylated, lipidated, non-lipidated, phosphorylated, non-phosphorylated, myristoylated, non-myristoylated, monomelic, multimeric, particulate, denatured, etc.).
Polypeptides used with the invention can be prepared by various means (e.g. recombinant expression, purification from cell culture, chemical synthesis, etc.). Recombinantly-expressed proteins are preferred, particularly for hybrid polypeptides. Polypeptides used with the invention are preferably provided in purified or substantially purified form i.e. substantially free from other polypeptides (e.g. free from naturally-occurring polypeptides), particularly from other staphylococcal or host cell polypeptides, and are generally at least about 50% pure (by weight), and usually at least about 90% pure i.e. less than about 50%, and more preferably less than about 10% (e.g. 5%) of a composition is made up of other expressed polypeptides. Thus the antigens in the compositions are separated from the whole organism with which the molecule is expressed.
Polypeptides used with the invention are preferably staphylococcal polypeptides.
The term "polypeptide" refers to amino acid polymers of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. Polypeptides can occur as single chains or associated chains.
The invention provides polypeptides comprising a sequence -P-Q- or -Q-P-, wherein: -P- is an amino acid sequence as defined above and -Q- is not a sequence as defined above i.e. the invention provides fusion proteins. Where the N-terminus codon of -P- is not ATG, but this codon is not present at the N-terminus of a polypeptide, it will be translated as the standard amino acid for that codon rather than as a Met. Where this codon is at the N-terminus of a polypeptide, however, it will be translated as Met. Examples of -Q- moieties include, but are not limited to, histidine tags (i.e. Hisπ where n = 3, 4, 5, 6, 7, 8, 9, 10 or more), maltose-binding protein, or glutathione-S-transferase (GST).
The invention also provides a process for producing a polypeptide of the invention, comprising the step of culturing a host cell transformed with nucleic acid of the invention under conditions which induce polypeptide expression. Although expression of the polypeptides of the invention may take place in a Staphylococcus, the invention will usually use a heterologous host for expression (recombinant expression). The heterologous host may be prokaryotic (e.g. a bacterium) or eukaryotic. It may be E.coli, but other suitable hosts include Bacillus subtilis, Vibrio cholerae, Salmonella typhi, Salmonella typhimurium, Neisseria lactamica, Neisseria cinerea, Mycobacteria (e.g. M.tuberculosis), yeasts, etc. Compared to the wild-type S. aureus genes encoding polypeptides of the invention, it is helpful to change codons to optimise expression efficiency in such hosts without affecting the encoded amino acids.
The invention provides a process for producing a polypeptide of the invention, comprising the step of synthesising at least part of the polypeptide by chemical means. Nucleic acids
The invention also provides nucleic acid encoding polypeptides and hybrid polypeptides of the invention. It also provides nucleic acid comprising a nucleotide sequence that encodes one or more polypeptides or hybrid polypeptides of the invention.
The invention also provides nucleic acid comprising nucleotide sequences having sequence identity to such nucleotide sequences. Identity between sequences is preferably determined by the Smith- Waterman homology search algorithm as described above. Such nucleic acids include those using alternative codons to encode the same amino acid.
The invention also provides nucleic acid which can hybridize to these nucleic acids. Hybridization reactions can be performed under conditions of different "stringency". Conditions that increase stringency of a hybridization reaction of widely known and published in the art (e.g. page 7.52 of reference 276). Examples of relevant conditions include (in order of increasing stringency): incubation temperatures of 25°C, 37°C, 50°C, 55°C and 68°C; buffer concentrations of 10 x SSC, 6 x SSC, 1 x SSC, 0.1 x SSC (where SSC is 0.15 M NaCl and 15 mM citrate buffer) and their equivalents using other buffer systems; formamide concentrations of 0%, 25%, 50%, and 75%; incubation times from 5 minutes to 24 hours; 1, 2, or more washing steps; wash incubation times of 1, 2, or 15 minutes; and wash solutions of 6 x SSC, 1 x SSC, 0.1 x SSC, or de-ionized water. Hybridization techniques and their optimization are well known in the art (e.g. see refs 75, 76, 276, 278, etc.].
In some embodiments, nucleic acid of the invention hybridizes to a target under low stringency conditions; in other embodiments it hybridizes under intermediate stringency conditions; in preferred embodiments, it hybridizes under high stringency conditions. An exemplary set of low stringency hybridization conditions is 50°C and 10 x SSC. An exemplary set of intermediate stringency hybridization conditions is 55°C and 1 x SSC. An exemplary set of high stringency hybridization conditions is 68°C and 0.1 x SSC. The invention includes nucleic acid comprising sequences complementary to these sequences (e.g. for antisense or probing, or for use as primers). Nucleic acids of the invention can be used in hybridisation reactions (e.g. Northern or Southern blots, or in nucleic acid microarrays or 'gene chips') and amplification reactions (e.g. PCR, SDA, SSSR, LCR, TMA, NASBA, etc.) and other nucleic acid techniques.
Nucleic acid according to the invention can take various forms (e.g. single-stranded, double-stranded, vectors, primers, probes, labelled etc.). Nucleic acids of the invention may be circular or branched, but will generally be linear. Unless otherwise specified or required, any embodiment of the invention that utilizes a nucleic acid may utilize both the double-stranded form and each of two complementary single-stranded forms which make up the double-stranded form. Primers and probes are generally single-stranded, as are antisense nucleic acids. Nucleic acids of the invention are preferably provided in purified or substantially purified form i.e. substantially free from other nucleic acids (e.g. free from naturally-occurring nucleic acids), particularly from other staphylococcal or host cell nucleic acids, generally being at least about 50% pure (by weight), and usually at least about 90% pure. Nucleic acids of the invention are preferably staphylococcal nucleic acids. Nucleic acids of the invention may be prepared in many ways e.g. by chemical synthesis (e.g. phosphoramidite synthesis of DNA) in whole or in part, by digesting longer nucleic acids using nucleases (e.g. restriction enzymes), by joining shorter nucleic acids or nucleotides (e.g. using ligases or polymerases), from genomic or cDNA libraries, etc.
Nucleic acid of the invention may be attached to a solid support (e.g. a bead, plate, filter, film, slide, microarray support, resin, etc.). Nucleic acid of the invention may be labelled e.g. with a radioactive or fluorescent label, or a biotin label. This is particularly useful where the nucleic acid is to be used in detection techniques e.g. where the nucleic acid is a primer or as a probe.
The term "nucleic acid" includes in general means a polymeric form of nucleotides of any length, which contain deoxyribonucleotides, ribonucleotides, and/or their analogs. It includes DNA, RNA, DNA/RNA hybrids. It also includes DNA or RNA analogs, such as those containing modified backbones (e.g. peptide nucleic acids (PNAs) or phosphorothioates) or modified bases. Thus the invention includes mRNA, tRNA, rRNA, ribozymes, DNA, cDNA, recombinant nucleic acids, branched nucleic acids, plasmids, vectors, probes, primers, etc. Where nucleic acid of the invention takes the form of RNA, it may or may not have a 5' cap. Nucleic acids of the invention may be part of a vector i.e. part of a nucleic acid construct designed for transduction/transfection of one or more cell types. Vectors may be, for example, "cloning vectors" which are designed for isolation, propagation and replication of inserted nucleotides, "expression vectors" which are designed for expression of a nucleotide sequence in a host cell, "viral vectors" which is designed to result in the production of a recombinant virus or virus-like particle, or "shuttle vectors", which comprise the attributes of more than one type of vector. Preferred vectors are plasmids. A "host cell" includes an individual cell or cell culture which can be or has been a recipient of exogenous nucleic acid. Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in total DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation and/or change. Host cells include cells transfected or infected in vivo or in vitro with nucleic acid of the invention. Where a nucleic acid is DNA, it will be appreciated that "U" in a RNA sequence will be replaced by "T" in the DNA. Similarly, where a nucleic acid is RNA, it will be appreciated that "T" in a DNA sequence will be replaced by "U" in the RNA.
The term "complement" or "complementary" when used in relation to nucleic acids refers to Watson- Crick base pairing. Thus the complement of C is G, the complement of G is C, the complement of A is T (or U), and the complement of T (or U) is A. It is also possible to use bases such as I (the purine inosine) e.g. to complement pyrimidines (C or T).
Nucleic acids of the invention can be used, for example: to produce polypeptides; as hybridization probes for the detection of nucleic acid in biological samples; to generate additional copies of the nucleic acids; to generate ribozymes or antisense oligonucleotides; as single-stranded DNA primers or probes; or as triple-strand forming oligonucleotides.
The invention provides a process for producing nucleic acid of the invention, wherein the nucleic acid is synthesised in part or in whole using chemical means.
The invention provides vectors comprising nucleotide sequences of the invention (e.g. cloning or expression vectors) and host cells transformed with such vectors. Nucleic acid amplification according to the invention may be quantitative and/or real-time.
For certain embodiments of the invention, nucleic acids are preferably at least 7 nucleotides in length (e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300 nucleotides or longer). For certain embodiments of the invention, nucleic acids are preferably at most 500 nucleotides in length (e.g. 450, 400, 350, 300, 250, 200, 150, 140, 130, 120, 110, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15 nucleotides or shorter).
Primers and probes of the invention, and other nucleic acids used for hybridization, are preferably between 10 and 30 nucleotides in length (e.g. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides).
Strains and variants
Antigens are defined above by reference to existing nomenclature (e.g. "CIfA"), to "sta" numbers or to "NW " numbers. Table 1 herein relates these three naming/numbering systems to existing SAOUHSC numbering and/or NWMN numbering. SAOUHSC numbering refers to the genome of S.aureus strain NCTC 8325 (sequenced by Oklahoma University Health Sciences Center and disclosed in GenBank as CP000253.1; GL87201381), and individual SAOUHSC numbers are given as "locus tag" entries in the genome sequence's "features" section. Similarly, NWMN numbering refers to the genome of S.aureus strain Newman (isolated in 1952 from a human infection, and having robust virulence phenotype) disclosed in GenBank as AP009351.1 (GI: 150373012) and individual NWMN numbers are given as "locus tag" entries in the genome sequence's "features" section. Functional annotations for each antigen are also given in the databases.
Table 1 also includes the GI number for each antigen of the invention. Thus an exemplary amino acid and nucleotide sequence for any of these antigens can easily be found in public sequence databases from the NCTC 8325 and/or Newman strain, but the invention is not limited to sequences from the
NCTC 8325 and Newman strains. Genome sequences of several other strains of S.aureus are available, including those of MRSA strains N315 and Mu50 [77],.MW2, N315, COL, MRSA252,
MSSA476, RF122, USA300 (very virulent), JHl and JH9. Standard search and alignment techniques can be used to identify in any of these (or other) further genome sequences the homolog of any particular sequence from the Newman or NCTC 8325 strain. Moreover, the available sequences from the Newman and NCTC 8325 strains can be used to design primers for amplification of homologous sequences from other strains. Thus the invention is not limited to these two strains, but rather encompasses such variants and homologs from other strains of S.aureus, as well as non-natural variants. In general, suitable variants of a particular SEQ ID NO include its allelic variants, its polymorphic forms, its homologs, its orthologs, its paralogs, its mutants, etc.
Thus, for instance, polypeptides used with the invention may, compared to the SEQ ID NO herein, include one or more {e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) amino acid substitutions, such as conservative substitutions (i.e. substitutions of one amino acid with another which has a related side chain). Genetically-encoded amino acids are generally divided into four families: (1) acidic i.e. aspartate, glutamate; (2) basic i.e. lysine, arginine, histidine; (3) non-polar i.e. alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar i.e. glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes classified jointly as aromatic amino acids. In general, substitution of single amino acids within these families does not have a major effect on the biological activity. The polypeptides may also include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) single amino acid deletions relative to the SEQ ID NO sequences. The polypeptides may also include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) insertions (e.g. each of 1, 2, 3, 4 or 5 amino acids) relative to the SEQ ID NO sequences.
Similarly, a polypeptide used with the invention may comprise an amino acid sequence that: is identical (i.e. 100% identical) to a sequence disclosed in the sequence listing; shares sequence identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) with a sequence disclosed in the sequence listing; has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (or more) single amino acid alterations (deletions, insertions, substitutions), which may be at separate locations or may be contiguous, as compared to the sequences of (a) or (b); and when aligned with a particular sequence from the sequence listing using a pairwise alignment algorithm, each moving window of x amino acids from N-terminus to C-terminus (such that for an alignment that extends to p amino acids, where p>x, there are p-x+1 such windows) has at least xy identical aligned amino acids, where: x is selected from 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200; y is selected from 0.50, 0.60, 0.70, 0.75, 0.80, 0.85, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99; and iϊ xy is not an integer then it is rounded up to the nearest integer. The preferred pairwise alignment algorithm is the Needleman-Wunsch global alignment algorithm [78], using default parameters {e.g. with Gap opening penalty = 10.0, and with Gap extension penalty = 0.5, using the EBLOSUM62 scoring matrix). This algorithm is conveniently implemented in the needle tool in the EMBOSS package [79].
Where hybrid polypeptides are used, the individual antigens within the hybrid {i.e. individual -X- moieties) may be from one or more strains. Where n=2, for instance, X2 may be from the same strain as Xi or from a different strain. Where
Figure imgf000096_0001
(iii) X1^X2=X3 (iv) Xi£X2£C3 or (v) Xi=X3^X2, etc.
Within group (c), deletions or substitutions may be at the N-terminus and/or C-terminus, or may be between the two termini. Thus a truncation is an example of a deletion. Truncations may involve deletion of up to 40 (or more) amino acids at the N-terminus and/or C-terminus. N-terminus truncation can remove leader peptides e.g. to facilitate recombinant expression in a heterologous host. C-terminus truncation can remove anchor sequences e.g. to facilitate recombinant expression in a heterologous host.
In general, when an antigen comprises a sequence that is not identical to a complete S. aureus sequence from the sequence listing {e.g. when it comprises a sequence listing with <100% sequence identity thereto, or when it comprises a fragment thereof) it is preferred in each individual instance that the antigen can elicit an antibody which recognises the respective complete S. aureus sequence.
Mutant bacteria
The invention also provides a S. aureus bacterium in which one or more of the antigens from the various antigen groups of the invention has/have been knocked out. Techniques for producing knockout bacteria are well known, and knockout S. aureus strains have been reported. A knockout mutation may be situated in the coding region of the gene or may lie within its transcriptional control regions {e.g. within its promoter). A knockout mutation will reduce the level of mRNA encoding the antigen to <1% of that produced by the wild-type bacterium, preferably <0.5%, more preferably <0.1%, and most preferably to 0%. The invention also provides a S. aureus in which one or more of the antigens from the various antigen groups of the invention has a mutation which inhibits its activity. The gene encoding the antigen will have a mutation that changes the encoded amino acid sequence. Mutation may involve deletion, substitution, and/or insertion, any of which may be involve one or more amino acids. The invention also provides a bacterium, such as a S.aureus bacterium, which hyper-expresses an antigen of the invention.
The invention also provides a bacterium, such as a S.aureus bacterium, that constitutively expresses an antigen of the invention. The invention also provides a meningococcus comprising a gene encoding an antigen of the invention, wherein the gene is under the control of an inducible promoter. Immunogenic compositions and medicaments
Immunogenic compositions of the invention may be useful as vaccines. Vaccines according to the invention may either be prophylactic (i.e. to prevent infection) or therapeutic (i.e. to treat infection), but will typically be prophylactic.
Compositions may thus be pharmaceutically acceptable. They will usually include components in addition to the antigens e.g. they typically include one or more pharmaceutical carrier(s) and/or excipient(s). A thorough discussion of such components is available in reference 273.
Compositions will generally be administered to a mammal in aqueous form. Prior to administration, however, the composition may have been in a non-aqueous form. For instance, although some vaccines are manufactured in aqueous form, then filled and distributed and administered also in aqueous form, other vaccines are lyophilised during manufacture and are reconstituted into an aqueous form at the time of use. Thus a composition of the invention may be dried, such as a lyophilised formulation.
The composition may include preservatives such as thiomersal or 2-phenoxyethanol. It is preferred, however, that the vaccine should be substantially free from (i.e. less than 5μg/ml) mercurial material e.g. thiomersal-free. Vaccines containing no mercury are more preferred. Preservative-free vaccines are particularly preferred.
To improve thermal stability, a composition may include a temperature protective agent. Further details of such agents are provided below.
To control tonicity, it is preferred to include a physiological salt, such as a sodium salt. Sodium chloride (NaCl) is preferred, which may be present at between 1 and 20 mg/ml e.g. about 10±2mg/ml NaCl. Other salts that may be present include potassium chloride, potassium dihydrogen phosphate, disodium phosphate dehydrate, magnesium chloride, calcium chloride, etc. Compositions will generally have an osmolality of between 200 mOsm/kg and 400 mθsm/kg, preferably between 240-360 mθsm/kg, and will more preferably fall within the range of 290-310 mOsm/kg.
Compositions may include one or more buffers. Typical buffers include: a phosphate buffer; a Tris buffer; a borate buffer; a succinate buffer; a histidine buffer (particularly with an aluminum hydroxide adjuvant); or a citrate buffer. Buffers will typically be included in the 5-2OmM range.
The pH of a composition will generally be between 5.0 and 8.1, and more typically between 6.0 and 8.0 e.g. 6.5 and 7.5, or between 7.0 and 7.8.
The composition is preferably sterile. The composition is preferably non-pyrogenic e.g. containing <1 EU (endotoxin unit, a standard measure) per dose, and preferably <0.1 EU per dose. The composition is preferably gluten free.
The composition may include material for a single immunisation, or may include material for multiple immunisations (i.e. a 'multidose' kit). The inclusion of a preservative is preferred in multidose arrangements. As an alternative (or in addition) to including a preservative in multidose compositions, the compositions may be contained in a container having an aseptic adaptor for removal of material.
Human vaccines are typically administered in a dosage volume of about 0.5ml, although a half dose (i.e. about 0.25ml) may be administered to children.
Immunogenic compositions of the invention may also comprise one or more immunoregulatory agents. Preferably, one or more of the immunoregulatory agents include one or more adjuvants. The adjuvants may include a THl adjuvant and/or a TH2 adjuvant, further discussed below.
Thus the invention provides an immunogenic composition comprising a combination of:
(1) one or more antigen(s) selected from the first, second, third and fourth antigen groups (as defined above); and (2) an adjuvant, such as an aluminium hydroxide adjuvant (for example, one or more antigens may be adsorbed to aluminium hydroxide).
For instance, the invention provides an immunogenic composition comprising a combination of a sta006 antigen and an adjuvant, such as an aluminium hydroxide adjuvant. Similarly, the invention provides an immunogenic composition comprising a combination of a staOl l antigen and an adjuvant, such as an aluminium hydroxide adjuvant. These compositions are ideally buffered e.g. with a histidine buffer.
Adjuvants which may be used in compositions of the invention include, but are not limited to: A. Mineral-containing compositions
Mineral containing compositions suitable for use as adjuvants in the invention include mineral salts, such as aluminium salts and calcium salts (or mixtures thereof). Calcium salts include calcium phosphate (e.g. the "CAP" particles disclosed in ref. 80). Aluminum salts include hydroxides, phosphates, sulfates, etc., with the salts taking any suitable form (e.g. gel, crystalline, amorphous, etc.). Adsorption to these salts is preferred (e.g. all antigens may be adsorbed). The mineral containing compositions may also be formulated as a particle of metal salt [81].
The adjuvants known as aluminum hydroxide and aluminum phosphate may be used. These names are conventional, but are used for convenience only, as neither is a precise description of the actual chemical compound which is present (e.g. see chapter 9 of reference 82)). The invention can use any of the "hydroxide" or "phosphate" adjuvants that are in general use as adjuvants. The adjuvants known as "aluminium hydroxide" are typically aluminium oxyhydroxide salts, which are usually at least partially crystalline. The adjuvants known as "aluminium phosphate" are typically aluminium hydroxyphosphates, often also containing a small amount of sulfate (i.e. aluminium hydroxyphosphate sulfate). They may be obtained by precipitation, and the reaction conditions and concentrations during precipitation influence the degree of substitution of phosphate for hydroxyl in the salt.
A fibrous morphology (e.g. as seen in transmission electron micrographs) is typical for aluminium hydroxide adjuvants. The pi of aluminium hydroxide adjuvants is typically about 11 i.e. the adjuvant itself has a positive surface charge at physiological pH. Adsorptive capacities of between 1.8-2.6 mg protein per mg Al+^ at pH 7.4 have been reported for aluminium hydroxide adjuvants.
Aluminium phosphate adjuvants generally have a PO4/ Al molar ratio between 0.3 and 1.2, preferably between 0.8 and 1.2, and more preferably 0.95+0.1. The aluminium phosphate will generally be amorphous, particularly for hydroxyphosphate salts. A typical adjuvant is amorphous aluminium hydroxyphosphate with POVAl molar ratio between 0.84 and 0.92, included at 0.6mg Al3+/ml. The aluminium phosphate will generally be particulate (e.g. plate-like morphology as seen in transmission electron micrographs). Typical diameters of the particles are in the range 0.5-20μm (e.g. about 5-10μm) after any antigen adsorption. Adsorptive capacities of between 0.7-1.5 mg protein per mg AT+* at pH 7.4 have been reported for aluminium phosphate adjuvants. The point of zero charge (PZC) of aluminium phosphate is inversely related to the degree of substitution of phosphate for hydroxyl, and this degree of substitution can vary depending on reaction conditions and concentration of reactants used for preparing the salt by precipitation. PZC is also altered by changing the concentration of free phosphate ions in solution (more phosphate = more acidic PZC) or by adding a buffer such as a histidine buffer (makes PZC more basic). Aluminium phosphates used according to the invention will generally have a PZC of between 4.0 and 7.0, more preferably between 5.0 and 6.5 e.g. about 5.7. As shown below, adsorption of S.aureus protein antigens (except IsdA, StaO19 and StaO73) to an aluminium hydroxide adjuvant is advantageous, particularly in a multi-protein combination (in which all antigens may be adsorbed). A histidine buffer can usefully be included in such adjuvanted compositions. Suspensions of aluminium salts used to prepare compositions of the invention may contain a buffer {e.g. a phosphate or a histidine or a Tris buffer), but this is not always necessary. The suspensions are preferably sterile and pyrogen-free. A suspension may include free aqueous phosphate ions e.g. present at a concentration between 1.0 and 2O mM, preferably between 5 and 15 mM, and more preferably about 10 mM. The suspensions may also comprise sodium chloride. The invention can use a mixture of both an aluminium hydroxide and an aluminium phosphate. In this case there may be more aluminium phosphate than hydroxide e.g. a weight ratio of at least 2:1 e.g. >5:l, >6:l, >7:l, >8:l, >9:l, efc.
The concentration of Al+^ in a composition for administration to a patient is preferably less than 10mg/ml e.g. <5 mg/ml, <4 mg/ml, <3 mg/ml, <2 mg/ml, <1 mg/ml, etc. A preferred range is between 0.3 and lmg/ml. A maximum of 0.85mg/dose is preferred.
B. Oil Emulsions
Oil emulsion compositions suitable for use as adjuvants in the invention include squalene-water emulsions, such as MF59 [Chapter 10 of ref. 82; see also ref. 83] (5% Squalene, 0.5% Tween 80, and 0.5% Span 85, formulated into submicron particles using a microfluidizer). Complete Freund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA) may also be used.
Various oil-in-water emulsion adjuvants are known, and they typically include at least one oil and at least one surfactant, with the oil(s) and surfactant(s) being biodegradable (metabolisable) and biocompatible. The oil droplets in the emulsion are generally less than 5μm in diameter, and ideally have a sub-micron diameter, with these small sizes being achieved with a microfiuidiser to provide stable emulsions. Droplets with a size less than 220nm are preferred as they can be subjected to filter sterilization.
The emulsion can comprise oils such as those from an animal (such as fish) or vegetable source. Sources for vegetable oils include nuts, seeds and grains. Peanut oil, soybean oil, coconut oil, and olive oil, the most commonly available, exemplify the nut oils. Jojoba oil can be used e.g. obtained from the jojoba bean. Seed oils include safflower oil, cottonseed oil, sunflower seed oil, sesame seed oil and the like. In the grain group, corn oil is the most readily available, but the oil of other cereal grains such as wheat, oats, rye, rice, teff, triticale and the like may also be used. 6-10 carbon fatty acid esters of glycerol and 1,2-propanediol, while not occurring naturally in seed oils, may be prepared by hydrolysis, separation and esterification of the appropriate materials starting from the nut and seed oils. Fats and oils from mammalian milk are metabolizable and may therefore be used in the practice of this invention. The procedures for separation, purification, saponification and other means necessary for obtaining pure oils from animal sources are well known in the art. Most fish contain metabolizable oils which may be readily recovered. For example, cod liver oil, shark liver oils, and whale oil such as spermaceti exemplify several of the fish oils which may be used herein. A number of branched chain oils are synthesized biochemically in 5-carbon isoprene units and are generally referred to as terpenoids. Shark liver oil contains a branched, unsaturated terpenoids known as squalene, 2,6,10,15, 19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, which is particularly preferred herein. Squalane, the saturated analog to squalene, is also a preferred oil. Fish oils, including squalene and squalane, are readily available from commercial sources or may be obtained by methods known in the art. Other preferred oils are the tocopherols (see below). Mixtures of oils can be used.
Surfactants can be classified by their 'HLB' (hydrophile/lipophile balance). Preferred surfactants of the invention have a HLB of at least 10, preferably at least 15, and more preferably at least 16. The invention can be used with surfactants including, but not limited to: the polyoxyethylene sorbitan esters surfactants (commonly referred to as the Tweens), especially polysorbate 20 and polysorbate 80; copolymers of ethylene oxide (EO), propylene oxide (PO), and/or butylene oxide (BO), sold under the DOWF AX™ tradename, such as linear EO/PO block copolymers; octoxynols, which can vary in the number of repeating ethoxy (oxy-l,2-ethanediyl) groups, with octoxynol-9 (Triton X-IOO, or t-octylphenoxypolyethoxyethanol) being of particular interest; (octylphenoxy)polyethoxyethanol (IGEPAL CA-630/NP-40); phospholipids such as phosphatidylcholine (lecithin); nonylphenol ethoxylates, such as the Tergitol™ NP series; polyoxyethylene fatty ethers derived from lauryl, cetyl, stearyl and oleyl alcohols (known as Brij surfactants), such as triethyleneglycol monolauryl ether (Brij 30); and sorbitan esters (commonly known as the SPANs), such as sorbitan trioleate (Span 85) and sorbitan monolaurate. Non-ionic surfactants are preferred. Preferred surfactants for including in the emulsion are Tween 80 (polyoxyethylene sorbitan monooleate), Span 85 (sorbitan trioleate), lecithin and Triton X- 100.
Mixtures of surfactants can be used e.g. Tween 80/Span 85 mixtures. A combination of a polyoxyethylene sorbitan ester such as polyoxyethylene sorbitan monooleate (Tween 80) and an octoxynol such as t-octylphenoxypolyethoxyethanol (Triton X-IOO) is also suitable. Another useful combination comprises laureth 9 plus a polyoxyethylene sorbitan ester and/or an octoxynol. Preferred amounts of surfactants (% by weight) are: polyoxyethylene sorbitan esters (such as Tween 80) 0.01 to 1%, in particular about 0.1 %; octyl- or nonylphenoxy polyoxyethanols (such as Triton X-100, or other detergents in the Triton series) 0.001 to 0.1 %, in particular 0.005 to 0.02%; polyoxyethylene ethers (such as laureth 9) 0.1 to 20 %, preferably 0.1 to 10 % and in particular 0.1 to 1 % or about 0.5%. Preferred emulsion adjuvants have an average droplets size of <1 μm e.g. <750nm, <500nm, <400nm, <300nm, <250nm, <220nm, <200nm, or smaller. These droplet sizes can conveniently be achieved by techniques such as microfluidisation. Specific oil-in-water emulsion adjuvants useful with the invention include, but are not limited to:
• A submicron emulsion of squalene, Tween 80, and Span 85. The composition of the emulsion by volume can be about 5% squalene, about 0.5% polysorbate 80 and about 0.5% Span 85. In weight terms, these ratios become 4.3% squalene, 0.5% polysorbate 80 and 0.48% Span 85. This adjuvant is known as 'MF59' [84-86], as described in more detail in Chapter 10 of ref. 87 and chapter 12 of ref. 88. The MF59 emulsion advantageously includes citrate ions e.g. 1OmM sodium citrate buffer.
• An emulsion of squalene, a tocopherol, and polysorbate 80 (Tween 80). The emulsion may include phosphate buffered saline. It may also include Span 85 (e.g. at 1%) and/or lecithin. These emulsions may have from 2 to 10% squalene, from 2 to 10% tocopherol and from 0.3 to
3% Tween 80, and the weight ratio of squalene:tocopherol is preferably <1 as this provides a more stable emulsion. Squalene and Tween 80 may be present volume ratio of about 5:2 or at a weight ratio of about 11:5. One such emulsion can be made by dissolving Tween 80 in PBS to give a 2% solution, then mixing 90ml of this solution with a mixture of (5g of DL-α-tocopherol and 5ml squalene), then microfiuidising the mixture. The resulting emulsion may have submicron oil droplets e.g. with an average diameter of between 100 and 250nm, preferably about 180nm. The emulsion may also include a 3-de-O-acylated monophosphoryl lipid A (3d-MPL). Another useful emulsion of this type may comprise, per human dose, 0.5-10 mg squalene, 0.5-11 mg tocopherol, and 0.1-4 mg polysorbate 80 [89]. • An emulsion of squalene, a tocopherol, and a Triton detergent (e.g. Triton X-100). The emulsion may also include a 3d-MPL (see below). The emulsion may contain a phosphate buffer.
• An emulsion comprising a polysorbate (e.g. polysorbate 80), a Triton detergent (e.g. Triton X-100) and a tocopherol (e.g. an α-tocopherol succinate). The emulsion may include these three components at a mass ratio of about 75:11:10 (e.g. 750μg/ml polysorbate 80, HOμg/ml
Triton X-100 and lOOμg/ml α-tocopherol succinate), and these concentrations should include any contribution of these components from antigens. The emulsion may also include squalene. The emulsion may also include a 3d-MPL (see below). The aqueous phase may contain a phosphate buffer. • An emulsion of squalane, polysorbate 80 and poloxamer 401 ("Pluronic™ L121"). The emulsion can be formulated in phosphate buffered saline, pH 7.4. This emulsion is a useful delivery vehicle for muramyl dipeptides, and has been used with threonyl-MDP in the "SAF-I" adjuvant [90] (0.05-1% Thr-MDP, 5% squalane, 2.5% Pluronic L121 and 0.2% polysorbate 80). It can also be used without the Thr-MDP, as in the "AF" adjuvant [91] (5% squalane, 1.25% Pluronic L121 and 0.2% polysorbate 80). Microfluidisation is preferred.
• An emulsion comprising squalene, an aqueous solvent, a polyoxyethylene alkyl ether hydrophilic nonionic surfactant (e.g. polyoxyethylene (12) cetostearyl ether) and a hydrophobic nonionic surfactant (e.g. a sorbitan ester or mannide ester, such as sorbitan monoleate or 'Span 80'). The emulsion is preferably thermoreversible and/or has at least 90% of the oil droplets (by volume) with a size less than 200 nm [92]. The emulsion may also include one or more of: alditol; a cryoprotective agent (e.g. a sugar, such as dodecylmaltoside and/or sucrose); and/or an alkylpolyglycoside. The emulsion may include a TLR4 agonist [93].
Such emulsions may be lyophilized.
• An emulsion of squalene, poloxamer 105 and Abil-Care [94]. The final concentration (weight) of these components in adjuvanted vaccines are 5% squalene, 4% poloxamer 105 (pluronic polyol) and 2% Abil-Care 85 (Bis-PEG/PPG-16/16 PEG/PPG-16/16 dimethicone; caprylic/capric triglyceride).
• An emulsion having from 0.5-50% of an oil, 0.1-10% of a phospholipid, and 0.05-5% of a non-ionic surfactant. As described in reference 95, preferred phospholipid components are phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, phosphatidic acid, sphingomyelin and cardiolipin. Submicron droplet sizes are advantageous.
• A submicron oil-in-water emulsion of a non-metabolisable oil (such as light mineral oil) and at least one surfactant (such as lecithin, Tween 80 or Span 80). Additives may be included, such as QuilA saponin, cholesterol, a saponin-lipophile conjugate (such as GPI-0100, described in reference 96, produced by addition of aliphatic amine to desacylsaponin via the carboxyl group of glucuronic acid), dimethyidioctadecylammonium bromide and/or N,N-dioctadecyl-N,N-bis
(2-hydroxyethyl)propanediamine.
• An emulsion in which a saponin (e.g. QuilA or QS21) and a sterol (e.g. a cholesterol) are associated as helical micelles [97].
• An emulsion comprising a mineral oil, a non-ionic lipophilic ethoxylated fatty alcohol, and a non-ionic hydrophilic surfactant (e.g. an ethoxylated fatty alcohol and/or polyoxyethylene- polyoxypropylene block copolymer) [98].
• An emulsion comprising a mineral oil, a non-ionic hydrophilic ethoxylated fatty alcohol, and a non-ionic lipophilic surfactant (e.g. an ethoxylated fatty alcohol and/or polyoxyethylene- polyoxypropylene block copolymer) [98]. In some embodiments an emulsion may be mixed with antigen extemporaneously, at the time of delivery, and thus the adjuvant and antigen may be kept separately in a packaged or distributed vaccine, ready for final formulation at the time of use. In other embodiments an emulsion is mixed with antigen during manufacture, and thus the composition is packaged in a liquid adjuvanted form,. The antigen will generally be in an aqueous form, such that the vaccine is finally prepared by mixing two liquids. The volume ratio of the two liquids for mixing can vary (e.g. between 5: 1 and 1 :5) but is generally about 1:1. Where concentrations of components are given in the above descriptions of specific emulsions, these concentrations are typically for an undiluted composition, and the concentration after mixing with an antigen solution will thus decrease.
Where a composition includes a tocopherol, any of the α, β, γ, δ, ε or ξ tocopherols can be used, but α-tocopherols are preferred. The tocopherol can take several forms e.g. different salts and/or isomers. Salts include organic salts, such as succinate, acetate, nicotinate, etc. D-α-tocopherol and
DL-α-tocopherol can both be used. Tocopherols are advantageously included in vaccines for use in elderly patients {e.g. aged 60 years or older) because vitamin E has been reported to have a positive effect on the immune response in this patient group [99]. They also have antioxidant properties that may help to stabilize the emulsions [100]. A preferred α-tocopherol is DL-α-tocopherol, and the preferred salt of this tocopherol is the succinate. The succinate salt has been found to cooperate with
TNF-related ligands in vivo.
C. Saponin formulations [chapter 22 ofref. 82]
Saponin formulations may also be used as adjuvants in the invention. Saponins are a heterogeneous group of sterol glycosides and triterpenoid glycosides that are found in the bark, leaves, stems, roots and even flowers of a wide range of plant species. Saponin from the bark of the Quillaia saponaria Molina tree have been widely studied as adjuvants. Saponin can also be commercially obtained from Smilax ornata (sarsaprilla), Gypsophilla paniculata (brides veil), and Saponaria officianalis (soap root). Saponin adjuvant formulations include purified formulations, such as QS21, as well as lipid formulations, such as ISCOMs. QS21 is marketed as Stimulon™. Saponin compositions have been purified using HPLC and RP-HPLC. Specific purified fractions using these techniques have been identified, including QS7, QS 17, QS 18, QS21, QH-A, QH-B and QH-C. Preferably, the saponin is QS21. A method of production of QS21 is disclosed in ref. 101. Saponin formulations may also comprise a sterol, such as cholesterol [102].
Combinations of saponins and cholesterols can be used to form unique particles called immunostimulating complexs (ISCOMs) [chapter 23 of ref. 82]. ISCOMs typically also include a phospholipid such as phosphatidylethanolamine or phosphatidylcholine. Any known saponin can be used in ISCOMs. Preferably, the ISCOM includes one or more of QuilA, QHA & QHC. ISCOMs are further described in refs. 102-104. Optionally, the ISCOMS may be devoid of additional detergent [105]. A review of the development of saponin based adjuvants can be found in refs. 106 & 107.
D. Virosomes and virus-like particles
Virosomes and virus-like particles (VLPs) can also be used as adjuvants in the invention. These structures generally contain one or more proteins from a virus optionally combined or formulated with a phospholipid. They are generally non-pathogenic, non-replicating and generally do not contain any of the native viral genome. The viral proteins may be recombinantly produced or isolated from whole viruses. These viral proteins suitable for use in virosomes or VLPs include proteins derived from influenza virus (such as HA or NA), Hepatitis B virus (such as core or capsid proteins), Hepatitis E virus, measles virus, Sindbis virus, Rotavirus, Foot-and-Mouth Disease virus, Retrovirus, Norwalk virus, human Papilloma virus, HIV, RNA-phages, Qβ-phage (such as coat proteins), GA- phage, fr-phage, AP205 phage, and Ty (such as retrotransposon Ty protein pi). VLPs are discussed further in refs. 108-113. Virosomes are discussed further in, for example, ref. 114
E. Bacterial or microbial derivatives
Adjuvants suitable for use in the invention include bacterial or microbial derivatives such as non-toxic derivatives of enterobacterial lipopolysaccharide (LPS), Lipid A derivatives, immunostimulatory oligonucleotides and ADP-ribosylating toxins and detoxified derivatives thereof. Non-toxic derivatives of LPS include monophosphoryl lipid A (MPL) and 3-O-deacylated MPL (3dMPL). 3dMPL is a mixture of 3 de-O-acylated monophosphoryl lipid A with 4, 5 or 6 acylated chains. A preferred "small particle" form of 3 De-O-acylated monophosphoryl lipid A is disclosed in ref. 115. Such "small particles" of 3dMPL are small enough to be sterile filtered through a 0.22μm membrane [115]. Other non-toxic LPS derivatives include monophosphoryl lipid A mimics, such as aminoalkyl glucosaminide phosphate derivatives e.g. RC-529 [ 116, 117] .
Lipid A derivatives include derivatives of lipid A from Escherichia coli such as OM- 174. OM- 174 is described for example in refs. 118 & 119.
Immunostimulatory oligonucleotides suitable for use as adjuvants in the invention include nucleotide sequences containing a CpG motif (a dinucleotide sequence containing an unmethylated cytosine linked by a phosphate bond to a guanosine). Double-stranded RNAs and oligonucleotides containing palindromic or poly(dG) sequences have also been shown to be immunostimulatory.
The CpG' s can include nucleotide modifications/analogs such as phosphorothioate modifications and can be double-stranded or single-stranded. References 120, 121 and 122 disclose possible analog substitutions e.g. replacement of guanosine with 2'-deoxy-7-deazaguanosine. The adjuvant effect of CpG oligonucleotides is further discussed in refs. 123-128.
The CpG sequence may be directed to TLR9, such as the motif GTCGTT or TTCGTT [129]. The CpG sequence may be specific for inducing a ThI immune response, such as a CpG-A ODN, or it may be more specific for inducing a B cell response, such a CpG-B ODN. CpG-A and CpG-B ODNs are discussed in refs. 130-132. Preferably, the CpG is a CpG-A ODN. Preferably, the CpG oligonucleotide is constructed so that the 5' end is accessible for receptor recognition. Optionally, two CpG oligonucleotide sequences may be attached at their 3' ends to form "immunomers". See, for example, refs. 129 & 133-135.
A useful CpG adjuvant is CpG7909, also known as ProMune™ (Coley Pharmaceutical Group, Inc.).
Another is CpGl 826. As an alternative, or in addition, to using CpG sequences, TpG sequences can be used [136], and these oligonucleotides may be free from unmethylated CpG motifs. The immunostimulatory oligonucleotide may be pyrimidine-rich. For example, it may comprise more than one consecutive thymidine nucleotide (e.g. TTTT, as disclosed in ref. 136), and/or it may have a nucleotide composition with >25% thymidine (e.g. >35%, >40%, >50%, >60%, >80%, etc.). For example, it may comprise more than one consecutive cytosine nucleotide (e.g. CCCC, as disclosed in ref. 136), and/or it may have a nucleotide composition with >25% cytosine (e.g. >35%, >40%, >50%, >60%, >80%, etc.). These oligonucleotides may be free from unmethylated CpG motifs. Immunostimulatory oligonucleotides will typically comprise at least 20 nucleotides. They may comprise fewer than 100 nucleotides.
A particularly useful adjuvant based around immunostimulatory oligonucleotides is known as IC-31™ [137]. Thus an adjuvant used with the invention may comprise a mixture of (i) an oligonucleotide (e.g. between 15-40 nucleotides) including at least one (and preferably multiple) CpI motifs (i.e. a cytosine linked to an inosine to form a dinucleotide), and (ii) a polycationic polymer, such as an oligopeptide (e.g. between 5-20 amino acids) including at least one (and preferably multiple) Lys-Arg-Lys tripeptide sequence(s). The oligonucleotide may be a deoxynucleotide comprising 26-mer sequence 5'-(IC)i3-3' (SEQ ID NO: 175). The polycationic polymer may be a peptide comprising 11-mer amino acid sequence KLKLLLLLKLK (SEQ ID NO: 176). The oligonucleotide and polymer can form complexes e.g. as disclosed in references 138 & 139.
Bacterial ADP-ribosylating toxins and detoxified derivatives thereof may be used as adjuvants in the invention. Preferably, the protein is derived from E.coli (E.coli heat labile enterotoxin "LT"), cholera ("CT"), or pertussis ("PT"). The use of detoxified ADP-ribosylating toxins as mucosal adjuvants is described in ref. 140 and as parenteral adjuvants in ref. 141. The toxin or toxoid is preferably in the form of a holotoxin, comprising both A and B subunits. Preferably, the A subunit contains a detoxifying mutation; preferably the B subunit is not mutated. Preferably, the adjuvant is a detoxified LT mutant such as LT-K63, LT-R72, and LT-Gl 92. The use of ADP-ribosylating toxins and detoxified derivatives thereof, particularly LT-K63 and LT-R72, as adjuvants can be found in refs. 142-149. A useful CT mutant is or CT-E29H [150]. Numerical reference for amino acid substitutions is preferably based on the alignments of the A and B subunits of ADP-ribosylating toxins set forth in ref. 151, specifically incorporated herein by reference in its entirety.
F. Human immunomodulators Human immunomodulators suitable for use as adjuvants in the invention include cytokines, such as interleukins (e.g. IL-I, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12 [152], etc.) [153], interferons (e.g. interferon-γ), macrophage colony stimulating factor, and tumor necrosis factor. A preferred immunomodulator is IL-12.
G. Bioadhesives and Mucoadhesives Bioadhesives and mucoadhesives may also be used as adjuvants in the invention. Suitable bioadhesives include esterified hyaluronic acid microspheres [154] or mucoadhesives such as cross-linked derivatives of poly(acrylic acid), polyvinyl alcohol, polyvinyl pyrollidone, polysaccharides and carboxymethylcellulose. Chitosan and derivatives thereof may also be used as adjuvants in the invention [155].
H. Microparticles
Microparticles may also be used as adjuvants in the invention. Microparticles (i.e. a particle of ~100nm to ~150μm in diameter, more preferably ~200nm to ~30μm in diameter, and most preferably
~500nm to ~10μm in diameter) formed from materials that are biodegradable and non-toxic (e.g. a poly(α-hydroxy acid), a polyhydroxybutyric acid, a polyorthoester, a polyanhydride, a polycaprolactone, etc.), with poly(lactide-co-glycolide) are preferred, optionally treated to have a negatively-charged surface (e.g. with SDS) or a positively-charged surface (e.g. with a cationic detergent, such as CTAB).
/. Liposomes (Chapters 13 & 14 ofref. 82)
Examples of liposome formulations suitable for use as adjuvants are described in refs. 156-158.
J. Polyoxyethylene ether and polyoxyethylene ester formulations
Adjuvants suitable for use in the invention include polyoxyethylene ethers and polyoxyethylene esters [159]. Such formulations further include polyoxyethylene sorbitan ester surfactants in combination with an octoxynol [160] as well as polyoxyethylene alkyl ethers or ester surfactants in combination with at least one additional non-ionic surfactant such as an octoxynol [161]. Preferred polyoxyethylene ethers are selected from the following group: polyoxyethylene-9-lauryl ether (laureth 9), polyoxyethylene-9-steoryl ether, polyoxytheylene-8-steoryl ether, polyoxyethylene-4- lauryl ether, polyoxyethylene-35-lauryl ether, and polyoxyethylene-23-lauryl ether.
K. Phosphazenes
A phosphazene, such as poly[di(carboxylatophenoxy)phosphazene] ("PCPP") as described, for example, in references 162 and 163, may be used.
L. Muramyl peptides Examples of muramyl peptides suitable for use as adjuvants in the invention include N-acetyl- muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP), and N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(r-2'-dipahnitoyl-5/j- glycero-3 -hydroxyphosphoryloxy)-ethylamine MTP-PE) .
M. Imidazoquinolone Compounds. Examples of imidazoquinolone compounds suitable for use adjuvants in the invention include Imiquimod ("R-837") [164,165], Resiquimod ("R-848") [166], and their analogs; and salts thereof (e.g. the hydrochloride salts). Further details about immunostimulatory imidazoquinolines can be found in references 167 to 171.
N. Substituted ureas Substituted ureas useful as adjuvants include compounds of formula I, II or III, or salts thereof: II III
Figure imgf000108_0001
as defined in reference 172, such as 'ER 803058', 'ER 803732', 'ER 804053', ER 804058', 'ER 804059', 'ER 804442', 'ER 804680', 'ER 804764', ER 803022 or 'ER 804057' e.g.:
Figure imgf000108_0002
O. Further adjuvants
Further adjuvants that may be used with the invention include:
• An aminoalkyl glucosaminide phosphate derivative, such as RC-529 [173, 174].
Cyclic diguanylate ('c-di-GMP'), which has been reported as a useful adjuvant for S.aureus vaccines [175].
A thiosemicarbazone compound, such as those disclosed in reference 176. Methods of formulating, manufacturing, and screening for active compounds are also described in reference 176. The thiosemicarbazones are particularly effective in the stimulation of human peripheral blood mononuclear cells for the production of cytokines, such as TNF-α.
A tryptanthrin compound, such as those disclosed in reference 177. Methods of formulating, manufacturing, and screening for active compounds are also described in reference 177. The thiosemicarbazones are particularly effective in the stimulation of human peripheral blood mononuclear cells for the production of cytokines, such as TNF-α.
• A nucleoside analog, such as: (a) Isatorabine (ANA-245; 7-thia-8-oxoguanosine):
Figure imgf000109_0001
and prodrugs thereof; (b) ANA975; (c) ANA-025-1; (d) ANA380; (e) the compounds disclosed in references 178 to 180Loxoribine (7-allyl-8-oxoguanosine) [181].
• Compounds disclosed in reference 182, including: Acylpiperazine compounds, Indoledione compounds, Tetrahydraisoquinoline (THIQ) compounds, Benzocyclodione compounds, Aminoazavinyl compounds, Aminobenzimidazole quinolinone (ABIQ) compounds [183,184], Hydrapthalamide compounds, Benzophenone compounds, Isoxazole compounds, Sterol compounds, Quinazilinone compounds, Pyrrole compounds [185], Anthraquinone compounds, Quinoxaline compounds, Triazine compounds, Pyrazalopyrimidine compounds, and Benzazole compounds [186].
• Compounds containing lipids linked to a phosphate-containing acyclic backbone, such as the TLR4 antagonist E5564 [187,188]: • A polyoxidonium polymer [ 189, 190] or other N-oxidized polyethylene-piperazine derivative.
• Methyl inosine 5 '-monophosphate ("MIMP") [191].
• A polyhydroxlated pyrrolizidine compound [192], such as one having formula:
Figure imgf000109_0002
where R is selected from the group comprising hydrogen, straight or branched, unsubstituted or substituted, saturated or unsaturated acyl, alkyl (e.g. cycloalkyl), alkenyl, alkynyl and aryl groups, or a pharmaceutically acceptable salt or derivative thereof. Examples include, but are not limited to: casuarine, casuarine-6-α-D-glucopyranose, 3-epj-casuarine, 7-ep/-casuarine, 3,7-diep/-casuarine, etc. • A CDId ligand, such as an α-glycosylceramide [193-200] (e.g. α-galactosylceramide), phytosphingosine-containing α-glycosylceramides, OCH, KRN7000 [(2S,3S,4R)-l-O-(α-D- galactopyranosyl)-2-(N-hexacosanoylamino)- 1 ,3,4-octadecanetriol], CRONY- 101, 3"-O- sulfo-galactosylceramide, etc.
• A gamma inulin [201] or derivative thereof, such as algammulin.
Figure imgf000110_0001
Adjuvant combinations
The invention may also comprise combinations of one or more of the adjuvants identified above. For example, the following adjuvant compositions may be used in the invention: (1) a saponin and an oil- in-water emulsion [202]; (2) a saponin (e.g. QS21) + a non-toxic LPS derivative (e.g. 3dMPL) [203]; (3) a saponin (e.g. QS21) + a non-toxic LPS derivative (e.g. 3dMPL) + a cholesterol; (4) a saponin (e.g. QS21) + 3dMPL + IL-12 (optionally + a sterol) [204]; (5) combinations of 3dMPL with, for example, QS21 and/or oil-in- water emulsions [205]; (6) SAF, containing 10% squalane, 0.4% Tween 80™, 5% pluronic-block polymer L121, and thr-MDP, either microfiuidized into a submicron emulsion or vortexed to generate a larger particle size emulsion. (7) Ribi™ adjuvant system (RAS), (Ribi Immunochem) containing 2% squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (Detox™); and (8) one or more mineral salts (such as an aluminum salt) + a non-toxic derivative of LPS (such as 3dMPL). Other substances that act as immunostimulating agents are disclosed in chapter 7 of ref. 82.
The use of an aluminium hydroxide and/or aluminium phosphate adjuvant is particularly preferred, and antigens are generally adsorbed to these salts. Calcium phosphate is another preferred adjuvant. Other preferred adjuvant combinations include combinations of ThI and Th2 adjuvants such as CpG & alum or resiquimod & alum. A combination of aluminium phosphate and 3dMPL may be used. The compositions of the invention may elicit both a cell mediated immune response as well as a humoral immune response. This immune response will preferably induce long lasting (e.g. neutralising) antibodies and a cell mediated immunity that can quickly respond upon exposure to pnuemococcus. Two types of T cells, CD4 and CD8 cells, are generally thought necessary to initiate and/or enhance cell mediated immunity and humoral immunity. CD8 T cells can express a CD8 co-receptor and are commonly referred to as Cytotoxic T lymphocytes (CTLs). CD8 T cells are able to recognized or interact with antigens displayed on MHC Class I molecules. CD4 T cells can express a CD4 co-receptor and are commonly referred to as T helper cells. CD4 T cells are able to recognize antigenic peptides bound to MHC class II molecules. Upon interaction with a MHC class II molecule, the CD4 cells can secrete factors such as cytokines. These secreted cytokines can activate B cells, cytotoxic T cells, macrophages, and other cells that participate in an immune response. Helper T cells or CD4+ cells can be further divided into two functionally distinct subsets: THl phenotype and TH2 phenotypes which differ in their cytokine and effector function.
Activated THl cells enhance cellular immunity (including an increase in antigen-specific CTL production) and are therefore of particular value in responding to intracellular infections. Activated THl cells may secrete one or more of IL-2, IFN-γ, and TNF-β. A THl immune response may result in local inflammatory reactions by activating macrophages, NK (natural killer) cells, and CD8 cytotoxic T cells (CTLs). A THl immune response may also act to expand the immune response by stimulating growth of B and T cells with IL-12. THl stimulated B cells may secrete IgG2a.
Activated TH2 cells enhance antibody production and are therefore of value in responding to extracellular infections. Activated TH2 cells may secrete one or more of IL-4, IL-5, IL-6, and IL-IO. A TH2 immune response may result in the production of IgGl, IgE, IgA and memory B cells for future protection.
An enhanced immune response may include one or more of an enhanced THl immune response and a TH2 immune response.
A THl immune response may include one or more of an increase in CTLs, an increase in one or more of the cytokines associated with a THl immune response (such as IL-2, IFN-γ, and TNF-β), an increase in activated macrophages, an increase in NK activity, or an increase in the production of
IgG2a. Preferably, the enhanced THl immune response will include an increase in IgG2a production.
A THl immune response may be elicited using a THl adjuvant. A THl adjuvant will generally elicit increased levels of IgG2a production relative to immunization of the antigen without adjuvant. THl adjuvants suitable for use in the invention may include for example saponin formulations, virosomes and virus like particles, non-toxic derivatives of enterobacterial lipopolysaccharide (LPS), immunostimulatory oligonucleotides. Immunostimulatory oligonucleotides, such as oligonucleotides containing a CpG motif, are preferred THl adjuvants for use in the invention.
A TH2 immune response may include one or more of an increase in one or more of the cytokines associated with a TH2 immune response (such as IL-4, IL-5, IL-6 and IL-IO), or an increase in the production of IgGl, IgE, IgA and memory B cells. Preferably, the enhanced TH2 immune resonse will include an increase in IgGl production.
A TH2 immune response may be elicited using a TH2 adjuvant. A TH2 adjuvant will generally elicit increased levels of IgGl production relative to immunization of the antigen without adjuvant. TH2 adjuvants suitable for use in the invention include, for example, mineral containing compositions, oil-emulsions, and ADP-ribosylating toxins and detoxified derivatives thereof. Mineral containing compositions, such as aluminium salts are preferred TH2 adjuvants for use in the invention.
Preferably, the invention includes a composition comprising a combination of a THl adjuvant and a TH2 adjuvant. Preferably, such a composition elicits an enhanced THl and an enhanced TH2 response, i.e., an increase in the production of both IgGl and IgG2a production relative to immunization without an adjuvant. Still more preferably, the composition comprising a combination of a THl and a TH2 adjuvant elicits an increased THl and/or an increased TH2 immune response relative to immunization with a single adjuvant {i.e., relative to immunization with a THl adjuvant alone or immunization with a TH2 adjuvant alone). The immune response may be one or both of a THl immune response and a TH2 response. Preferably, immune response provides for one or both of an enhanced THl response and an enhanced TH2 response.
The enhanced immune response may be one or both of a systemic and a mucosal immune response. Preferably, the immune response provides for one or both of an enhanced systemic and an enhanced mucosal immune response. Preferably the mucosal immune response is a TH2 immune response. Preferably, the mucosal immune response includes an increase in the production of IgA.
S.aureus infections can affect various areas of the body and so the compositions of the invention may be prepared in various forms. For example, the compositions may be prepared as injectables, either as liquid solutions or suspensions. Solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection can also be prepared {e.g. a lyophilised composition or a spray-freeze dried composition). The composition may be prepared for topical administration e.g. as an ointment, cream or powder. The composition may be prepared for oral administration e.g. as a tablet or capsule, as a spray, or as a syrup (optionally flavoured). The composition may be prepared for pulmonary administration e.g. as an inhaler, using a fine powder or a spray. The composition may be prepared as a suppository or pessary. The composition may be prepared for nasal, aural or ocular administration e.g. as drops. The composition may be in kit form, designed such that a combined composition is reconstituted just prior to administration to a patient. Such kits may comprise one or more antigens in liquid form and one or more lyophilised antigens.
Where a composition is to be prepared extemporaneously prior to use {e.g. where a component is presented in lyophilised form) and is presented as a kit, the kit may comprise two vials, or it may comprise one ready-filled syringe and one vial, with the contents of the syringe being used to reactivate the contents of the vial prior to injection.
Immunogenic compositions used as vaccines comprise an immunologically effective amount of antigen(s), as well as any other components, as needed. By 'immunologically effective amount', it is meant that the administration of that amount to an individual, either in a single dose or as part of a series, is effective for treatment or prevention. This amount varies depending upon the health and physical condition of the individual to be treated, age, the taxonomic group of individual to be treated (e.g. non-human primate, primate, etc.), the capacity of the individual's immune system to synthesise antibodies, the degree of protection desired, the formulation of the vaccine, the treating doctor's assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials. Where more than one antigen is included in a composition then two antigens may be present at the same dose as each other or at different doses.
As mentioned above, a composition may include a temperature protective agent, and this component may be particularly useful in adjuvanted compositions (particularly those containing a mineral adjuvant, such as an aluminium salt). As described in reference 206, a liquid temperature protective agent may be added to an aqueous vaccine composition to lower its freezing point e.g. to reduce the freezing point to below 0°C. Thus the composition can be stored below 0°C, but above its freezing point, to inhibit thermal breakdown. The temperature protective agent also permits freezing of the composition while protecting mineral salt adjuvants against agglomeration or sedimentation after freezing and thawing, and may also protect the composition at elevated temperatures e.g. above 400C. A starting aqueous vaccine and the liquid temperature protective agent may be mixed such that the liquid temperature protective agent forms from 1-80% by volume of the final mixture. Suitable temperature protective agents should be safe for human administration, readily miscible/soluble in water, and should not damage other components (e.g. antigen and adjuvant) in the composition. Examples include glycerin, propylene glycol, and/or polyethylene glycol (PEG). Suitable PEGs may have an average molecular weight ranging from 200-20,000 Da. In a preferred embodiment, the polyethylene glycol can have an average molecular weight of about 300 Da ('PEG-300').
The invention provides an immunogenic composition comprising: (i) one or more antigen(s) selected from the first, second, third or fourth antigen groups; and (ii) a temperature protective agent. This composition may be formed by mixing (i) an aqueous composition comprising one or more antigen(s) selected from the first, second, third or fourth antigen groups, with (ii) a temperature protective agent. The mixture may then be stored e.g. below 00C, from 0-20°C, from 20-35 °C, from
35-55°C, or higher. It may be stored in liquid or frozen form. The mixture may be lyophilised. The composition may alternatively be formed by mixing (i) a dried composition comprising one or more antigen(s) selected from the first, second, third or fourth antigen groups, with (ii) a liquid composition comprising the temperature protective agent. Thus component (ii) can be used to reconstitute component (i).
Methods of treatment, and administration of the vaccine
The invention also provides a method for raising an immune response in a mammal comprising the step of administering an effective amount of a composition of the invention. The immune response is preferably protective and preferably involves antibodies and/or cell-mediated immunity. The method may raise a booster response.
The invention also provides at least two antigens of the invention for combined use as a medicament e.g. for use in raising an immune response in a mammal. The invention also provides the use of at least two antigens of the invention in the manufacture of a medicament for raising an immune response in a mammal.
By raising an immune response in the mammal by these uses and methods, the mammal can be protected against S.aureus infection, including a nosocomial infection. More particularly, the mammal may be protected against a skin infection, pneumonia, meningitis, osteomyelitis endocarditis, toxic shock syndrome, and/or septicaemia.
The invention also provides a kit comprising a first component and a second component wherein neither the first component nor the second component is a composition of the invention as described above, but wherein the first component and the second component can be combined to provide a composition of the invention as described above. The kit may further include a third component comprising one or more of the following: instructions, syringe or other delivery device, adjuvant, or pharmaceutically acceptable formulating solution.
The invention also provides a delivery device pre-filled with an immunogenic composition of the invention.
The mammal is preferably a human. Where the vaccine is for prophylactic use, the human is preferably a child (e.g. a toddler or infant) or a teenager; where the vaccine is for therapeutic use, the human is preferably a teenager or an adult. A vaccine intended for children may also be administered to adults e.g. to assess safety, dosage, immunogenicity, etc. Other mammals which can usefully be immunised according to the invention are cows, dogs, horses, and pigs.
One way of checking efficacy of therapeutic treatment involves monitoring S.aureus infection after administration of the compositions of the invention. One way of checking efficacy of prophylactic treatment involves monitoring immune responses, systemically (such as monitoring the level of IgGl and IgG2a production) and/or mucosally (such as monitoring the level of IgA production), against the antigens in the compositions of the invention after administration of the composition. Typically, antigen-specific serum antibody responses are determined post-immunisation but pre-challenge whereas antigen-specific mucosal antibody responses are determined post-immunisation and post- challenge.
Another way of assessing the immunogenicity of the compositions of the present invention is to express the proteins recombinantly for screening patient sera or mucosal secretions by immunoblot and/or microarrays. A positive reaction between the protein and the patient sample indicates that the patient has mounted an immune response to the protein in question. This method may also be used to identify immunodominant antigens and/or epitopes within antigens.
The efficacy of vaccine compositions can also be determined in vivo by challenging animal models of S. aureus infection, e.g., guinea pigs or mice, with the vaccine compositions, hi particular, there are three useful animal models for the study of S. aureus infectious disease, namely: (i) the murine abscess model [207], (ii) the murine lethal infection model [207] and (iii) the murine pneumonia model [208]. The abscess model looks at abscesses in mouse kidneys after intravenous challenge. The lethal infection model looks at the number of mice which survive after being infected by a normally-lethal dose of S. aureus by the intravenous or intraperitoneal route. The pneumonia model also looks at the survival rate, but uses intranasal infection. A useful vaccine may be effective in one or more of these models. For instance, for some clinical situations it may be desirable to protect against pneumonia, without needing to prevent hematic spread or to promote opsonisation; in other situations the main desire may be to prevent hematic spread. Different antigens, and different antigen combinations, may contribute to different aspects of an effective vaccine. Compositions of the invention will generally be administered directly to a patient. Direct delivery may be accomplished by parenteral injection {e.g. subcutaneously, intraperitoneally, intravenously, intramuscularly, or to the interstitial space of a tissue), or mucosally, such as by rectal, oral (e.g. tablet, spray), vaginal, topical, transdermal or transcutaneous, intranasal, ocular, aural, pulmonary or other mucosal administration. The invention may be used to elicit systemic and/or mucosal immunity, preferably to elicit an enhanced systemic and/or mucosal immunity.
Preferably the enhanced systemic and/or mucosal immunity is reflected in an enhanced THl and/or TH2 immune response. Preferably, the enhanced immune response includes an increase in the production of IgGl and/or IgG2a and/or IgA. Dosage can be by a single dose schedule or a multiple dose schedule. Multiple doses may be used in a primary immunisation schedule and/or in a booster immunisation schedule, hi a multiple dose schedule the various doses may be given by the same or different routes e.g. a parenteral prime and mucosal boost, a mucosal prime and parenteral boost, etc. Multiple doses will typically be administered at least 1 week apart {e.g. about 2 weeks, about 3 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 16 weeks, etc.). Vaccines prepared according to the invention may be used to treat both children and adults. Thus a human patient may be less than 1 year old, 1-5 years old, 5-15 years old, 15-55 years old, or at least 55 years old. Preferred patients for receiving the vaccines are the elderly (e.g. >50 years old, >60 years old, and preferably >65 years), the young (e.g. <5 years old), hospitalised patients, healthcare workers, armed service and military personnel, pregnant women, the chronically ill, or immunodefϊcient patients. The vaccines are not suitable solely for these groups, however, and may be used more generally in a population.
Vaccines produced by the invention may be administered to patients at substantially the same time as (e.g. during the same medical consultation or visit to a healthcare professional or vaccination centre) other vaccines e.g. at substantially the same time as an influenza vaccine, a measles vaccine, a mumps vaccine, a rubella vaccine, a MMR vaccine, a varicella vaccine, a MMRV vaccine, a diphtheria vaccine, a tetanus vaccine, a pertussis vaccine, a DTP vaccine, a conjugated H.influenzae type b vaccine, an inactivated poliovirus vaccine, a hepatitis B virus vaccine, a meningococcal conjugate vaccine (such as a tetravalent A-C-Wl 35-Y vaccine), a respiratory syncytial virus vaccine, etc. Further non-staphylococcal vaccines suitable for co-administration may include one or more antigens listed on pages 33-46 of reference 51.
Nucleic acid immunisation
The immunogenic compositions described above include polypeptide antigens from S. aureus. In all cases, however, the polypeptide antigens can be replaced by nucleic acids (typically DNA) encoding those polypeptides, to give compositions, methods and uses based on nucleic acid immunisation. Nucleic acid immunisation is now a developed field (e.g. see references 209 to 216 etc.).
The nucleic acid encoding the immunogen is expressed in vivo after delivery to a patient and the expressed immunogen then stimulates the immune system. The active ingredient will typically take the form of a nucleic acid vector comprising: (i) a promoter; (ii) a sequence encoding the immunogen, operably linked to the promoter; and optionally (iii) a selectable marker. Preferred vectors may further comprise (iv) an origin of replication; and (v) a transcription terminator downstream of and operably linked to (ii). In general, (i) & (v) will be eukaryotic and (iii) & (iv) will be prokaryotic.
Preferred promoters are viral promoters e.g. from cytomegalovirus (CMV). The vector may also include transcriptional regulatory sequences (e.g. enhancers) in addition to the promoter and which interact functionally with the promoter. Preferred vectors include the immediate-early CMV enhancer/promoter, and more preferred vectors also include CMV intron A. The promoter is operably linked to a downstream sequence encoding an immunogen, such that expression of the immunogen-encoding sequence is under the promoter's control. Where a marker is used, it preferably functions in a microbial host (e.g. in a prokaryote, in a bacteria, in a yeast). The marker is preferably a prokaryotic selectable marker (e.g. transcribed under the control of a prokaryotic promoter). For convenience, typical markers are antibiotic resistance genes.
The vector of the invention is preferably an autonomously replicating episomal or extrachromosomal vector, such as a plasmid.
The vector of the invention preferably comprises an origin of replication. It is preferred that the origin of replication is active in prokaryotes but not in eukaryotes.
Preferred vectors thus include a prokaryotic marker for selection of the vector, a prokaryotic origin of replication, but a eukaryotic promoter for driving transcription of the immunogen-encoding sequence. The vectors will therefore (a) be amplified and selected in prokaryotic hosts without polypeptide expression, but (b) be expressed in eukaryotic hosts without being amplified. This arrangement is ideal for nucleic acid immunization vectors.
The vector of the invention may comprise a eukaryotic transcriptional terminator sequence downstream of the coding sequence. This can enhance transcription levels. Where the coding sequence does not have its own, the vector of the invention preferably comprises a polyadenylation sequence. A preferred polyadenylation sequence is from bovine growth hormone.
The vector of the invention may comprise a multiple cloning site
In addition to sequences encoding the immunogen and a marker, the vector may comprise a second eukaryotic coding sequence. The vector may also comprise an IRES upstream of said second sequence in order to permit translation of a second eukaryotic polypeptide from the same transcript as the immunogen. Alternatively, the immunogen-coding sequence may be downstream of an IRES.
The vector of the invention may comprise unmethylated CpG motifs e.g. unmethylated DNA sequences which have in common a cytosine preceding a guanosine, flanked by two 5' purines and two 3' pyrimidines. In their unmethylated form these DNA motifs have been demonstrated to be potent stimulators of several types of immune cell.
Vectors may be delivered in a targeted way. Receptor-mediated DNA delivery techniques are described in, for example, references 217 to 222. Therapeutic compositions containing a nucleic acid are administered in a range of about lOOng to about 200mg of DNA for local administration in a gene therapy protocol. Concentration ranges of about 500 ng to about 50 mg, about lμg to about 2 mg, about 5μg to about 500μg, and about 20μg to about lOOμg of DNA can also be used during a gene therapy protocol. Factors such as method of action (e.g. for enhancing or inhibiting levels of the encoded gene product) and efficacy of transformation and expression are considerations which will affect the dosage required for ultimate efficacy. Where greater expression is desired over a larger area of tissue, larger amounts of vector or the same amounts re-administered in a successive protocol of administrations, or several administrations to different adjacent or close tissue portions may be required to effect a positive therapeutic outcome. In all cases, routine experimentation in clinical trials will determine specific ranges for optimal therapeutic effect.
Vectors can be delivered using gene delivery vehicles. The gene delivery vehicle can be of viral or non-viral origin (see generally references 223 to 226). Viral-based vectors for delivery of a desired nucleic acid and expression in a desired cell are well known in the art. Exemplary viral-based vehicles include, but are not limited to, recombinant retroviruses (e.g. references 227 to 237), alphavirus-based vectors (e.g. Sindbis virus vectors, Semliki forest virus (ATCC VR-67; ATCC VR-1247), Ross River virus (ATCC VR-373; ATCC VR-1246) and Venezuelan equine encephalitis virus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-532); hybrids or chimeras of these viruses may also be used), poxvirus vectors (e.g. vaccinia, fowlpox, canarypox, modified vaccinia Ankara, etc.), adenovirus vectors, and adeno- associated virus (AAV) vectors (e.g. see refs. 238 to 243). Administration of DNA linked to killed adenovirus [244] can also be employed.
Non-viral delivery vehicles and methods can also be employed, including, but not limited to, polycationic condensed DNA linked or unlinked to killed adenovirus alone [e.g. 244], ligand-linked
DNA [245], eukaryotic cell delivery vehicles cells [e.g. refs. 246 to 250] and nucleic charge neutralization or fusion with cell membranes. Naked DNA can also be employed. Exemplary naked
DNA introduction methods are described in refs. 251 and 252. Liposomes (e.g. immunoliposomes) that can act as gene delivery vehicles are described in refs. 253 to 257. Additional approaches are described in references 258 & 259.
Further non- viral delivery suitable for use includes mechanical delivery systems such as the approach described in ref. 259. Moreover, the coding sequence and the product of expression of such can be delivered through deposition of photopolymerized hydrogel materials or use of ionizing radiation [e.g. refs. 260 & 261]. Other conventional methods for gene delivery that can be used for delivery of the coding sequence include, for example, use of hand-held gene transfer particle gun [262] or use of ionizing radiation for activating transferred genes [260 & 261].
Delivery DNA using PLG (poly(lactide-co-glycolide)} microparticles is a particularly preferred method e.g. by adsorption to the microparticles, which are optionally treated to have a negatively- charged surface (e.g. treated with SDS) or a positively-charged surface (e.g. treated with a cationic detergent, such as CTAB).
S. epidermidis
Although the invention focuses on S. aureus, the inventors also realise that the sta006 and staOl l antigens have homologs in S.epidermidis. For example, SEQ ID NO: 234 is the 'iron (Fe+3) ABC superfamily ATP binding cassette transporter, binding protein' from S.epidermidis strain M23864:W1, with 73% identity to SEQ ID NO: 42 (staOOό), and SEQ ID NO: 235 is the 'putative lipoprotein' from S.epidermidis strain RP62A, with 67% identity to SEQ ID NO: 47 (staOl 1). S.epidermidis is commonly present on human skin and can sometimes cause illness. Infection is usually associated with medical devices, such as catheters, and is a cause of nosocomial infections. The results disclosed herein for sta006 and staOl l against S. aureus suggest that the homologous proteins in S.epidermidis could be useful for immunising against this pathogen. The invention provides an immunogenic composition comprising:
(i) a polypeptide comprising an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 234; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 234, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more); and/or
(ii) a polypeptide comprising an amino acid sequence: (a) having 50% or more identity (e.g.
60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 235; and/or (b) comprising a fragment of at least 'n' consecutive amino acids of SEQ ID NO: 235, wherein 'n' is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50,
60, 70, 80, 90, 100, 150, 200, 250 or more).
The composition may also include an adjuvant. These compositions are particularly useful for immunising a mammal (including a human) against S.epidermidis infection.
Preferred fragments of (b) comprise an epitope from SEQ ID NO: 234 or 235, respectively. Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 234/235 while retaining at least one epitope of SEQ ID NO: 234/235.
More generally, the invention provides the use of the sta006 and/or staOl 1 homolog from any Staphylococcus species for immunising a mammal against that species.
Antibodies
Antibodies against S.aureus antigens can be used for passive immunisation. Thus the invention provides an antibody which is specific for an antigen in the first, second, third or fourth antigen groups. The invention also provides the use of such antibodies in therapy. The invention also provides the use of such antibodies in the manufacture of a medicament. The invention also provides a method for treating a mammal comprising the step of administering an effective amount of an antibody of the invention. As described above for immunogenic compositions, these methods and uses allow a mammal to be protected against S.aureus infection. The term "antibody" includes intact immunoglobulin molecules, as well as fragments thereof which are capable of binding an antigen. These include hybrid (chimeric) antibody molecules [263, 264]; F(ab')2 and F(ab) fragments and Fv molecules; non-covalent heterodimers [265, 266]; single-chain Fv molecules (sFv) [267]; dimeric and trimeric antibody fragment constructs; minibodies [268, 269]; humanized antibody molecules [270-272]; and any functional fragments obtained from such molecules, as well as antibodies obtained through non-conventional processes such as phage display. Preferably, the antibodies are monoclonal antibodies. Methods of obtaining monoclonal antibodies are well known in the art. Humanised or fully-human antibodies are preferred.
General The practice of the present invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular biology, immunology and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., references 273-280, etc.
"GI" numbering is used above. A GI number, or "Genlnfo Identifier", is a series of digits assigned consecutively to each sequence record processed by NCBI when sequences are added to its databases. The GI number bears no resemblance to the accession number of the sequence record. When a sequence is updated (e.g. for correction, or to add more annotation or information) then it receives a new GI number. Thus the sequence associated with a given GI number is never changed.
Where the invention concerns an "epitope", this epitope may be a B-cell epitope and/or a T-cell epitope. Such epitopes can be identified empirically (e.g. using PEPSCAN [281,282] or similar methods), or they can be predicted (e.g. using the Jameson- Wolf antigenic index [283], matrix-based approaches [284], MAPITOPE [285], TEPITOPE [286,287], neural networks [288], OptiMer & EpiMer [289, 290], ADEPT [291], Tsites [292], hydrophilicity [293], antigenic index [294] or the methods disclosed in references 295-299, etc.). Epitopes are the parts of an antigen that are recognised by and bind to the antigen binding sites of antibodies or T-cell receptors, and they may also be referred to as "antigenic determinants".
Where an antigen "domain" is omitted, this may involve omission of a signal peptide, of a cytoplasmic domain, of a transmembrane domain, of an extracellular domain, etc.
The term "comprising" encompasses "including" as well as "consisting" e.g. a composition "comprising" X may consist exclusively of X or may include something additional e.g. X + Y. The term "about" in relation to a numerical value JC is optional and means, for example, jc+10%.
References to a percentage sequence identity between two amino acid sequences means that, when aligned, that percentage of amino acids are the same in comparing the two sequences. This alignment and the percent homology or sequence identity can be determined using software programs known in the art, for example those described in section 7.7.18 of ref. 300. A preferred alignment is determined by the Smith- Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62. The Smith- Waterman homology search algorithm is disclosed in ref. 301.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 shows bacterial counts (Log cfu/ml) after challenge of mice previously immunised with the indicated antigens.
Figures 2 to 4 show survival (%) after challenge of mice previously immunised with various mixtures of antigens over 14 days. In Figure 2, the six groups from SA-10-a are, from top to bottom at day 14. groups (i), (iii) & (iv) together, (ii), IsdB, then the negative control, hi Figure 3, the six groups from SA-10-a are, from top to bottom at day 14. groups (i), (iii) & (iv) together, (ii), IsdB, then the negative control, hi Figure 3, the six groups from SA-10-b are, from top to bottom at day 14. groups (iii), (i), (iv), (ii) and IsdB together, then the negative control, hi Figure 4, the six groups from SA-14 are, from top to bottom at day 14. groups (iv), (ii), (i), (iii), negative control, and IsdB.
Figure 5 shows collected data on mouse survival from four different experiments after challenge of mice previously immunised with various compositions (PBS negative control; IsdB antigen; and "Combo- 1" and "Combo-2" antigen combinations of the invention). Individual symbols show the survival duration of individual mice; the horizontal bar for each group shows the median survival duration; the percentage figures are survival 14 days after challenge; and the p values at the top are t-Test comparisons of median survival durations between groups.
Figure 6 shows the number of colony forming units (cfu) in mouse kidneys after infection with 9x106 cfu of Newman strain in the abscess model. Horizontal bars are averages per group, and the figure beneath each group is the log reduction relative to the PBS control group.
Figure 7 shows bacterial count (log CFU/ml) in kidneys of mice in an abscess model experiment. Mice were challenged with the following strains: (A) MW2; (B) LAC; (C) Staphl9; or (D) MU50. Each point is an individual animal and the bar shows the median count per group. Mice had been immunised as shown on the x-axis label.
Figure 8 shows the formation of StaOl l oligomers in the presence of increasing concentrations of Ca+* ions. Numbers indicate mM concentrations, and a * indicates the presence of 5OmM EDTA.
Figure 9 shows IgG titers against (A) EsxAB (B) Sta006 (C) Hla-H35L (D) StaOl 1. Each graph has three groups, with a pair of bars per group. The right-hand bar in a pair shows pre-immune IgG and the left-hand bar shows post-immune IgG. The three groups are the compositions used for immunising and, from left to right, are: negative control of adjuvant alone; the Combo 1 combination; and the relevant antigen alone.
Figure 10 shows bacterial counts values (log CFU/ml) in mice after challenge with the indicated strains. Each point is an individual animal and the bar shows the median CFU. The P value beneath the IsdB and Combo columns is a comparison against the adjuvant-only control. Figure 11 shows the area of abscesses (mm2) in mice after challenge with Newman strain.
Figure 12 shows days of survival of mice after challenge with four different strains: Newman (o), ST-80 (D), USA300-FPR3757 (Δ) or USA300-Lac (x) strains. Each point is an individual animal, the bar shows the median survival, and the heading number shows the % of animals surviving after 15 days. Mice received aluminium hydroxide adjuvant alone, IsdB or Combo 1.
Figure 13 shows the median survival (days) of mice after challenge. The mice had been immunised with the antigens indicated on the X-axis. Each point is an individual animal and the bar shows the median survival. The heading numbers show the % of animals surviving after 15 days.
MODES FOR CARRYING OUT THE INVENTION Antigen selection
S.aureus proteins have been selected for use as vaccine components based on various criteria.
IsdA is a surface protein involved in iron uptake. It is detectable with a high molecular weight (>250kDa) in immunoblots of whole cell lysates and cell wall fractions of S. aureus. Furthermore, labelled anti-IsdA antibodies revealed extracellular structures. These structures were seen in a variety of growth and infection conditions, including iron positive conditions (in which IsdA expression is reported to be suppressed). The structures have a tail up to 4μm long, with a typical orientation parallel to the mammalian cell surface. Detached IsdA-positive structures were observed to adhere on the surface of epithelial cells, but lose cell junction localization. Epithelia/bacteria interaction may stimulate expression of the structures. In addition, the inventors have found that IsdA is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. Iron uptake is important for virulence, so the protein is likely to be available for immune attack at pathological stages of the bacterial life cycle. The inventors have found that the protein is not cytotoxic to human cells (see below). The protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
EsxA and EsxB are small acidic dimeric secreted proteins. The inventors have found that EsxA is highly conserved between different strains (present in 36/36 strains tested; see below), while EsxB is present in 25/36 strains. The proteins are involved in persisting an infection and so are likely to be available for immune attack at pathological stages of the bacterial life cycle. The inventors have found that a fusion of EsxA and EsxB ('EsxAB') is not cytotoxic to human cells (see below). It can also adsorb well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. Thus the antigens are useful for providing an immune response to prevent hematic spread of the bacterium. HIa is a pore-forming secreted toxin. This protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. It is an important virulence factor so is likely to be available for immune attack at pathological stages of the bacterial life cycle. It is not cytotoxic to human cells (see below). The protein can adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent pneumonia.
Spa is a surface protein involved in Fc binding. The inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested), thus offering protection across a broad population of circulating strains. It is important for virulence so is likely to be available for immune attack at pathological stages of the bacterial life cycle. The protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium. Sta006 (also known as FhuD2) is a surface protein involved in iron uptake. The inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. The inventors have found that the protein is not cytotoxic to human cells (see below). The protein can also adsorb well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium.
StaOl 1 is a surface lipoprotein. The inventors have found that this protein is well conserved between different strains (present in 36/36 strains tested; see below), thus offering protection across a broad population of circulating strains. The inventors have found that the protein is not cytotoxic to human cells (see below). The protein can also adsorb reasonably well to aluminium hydroxide (see below), which is useful for stable formulation for delivery to humans. It is useful for providing an immune response to prevent hematic spread of the bacterium. This protein has been shown to assemble into oligomers in the presence of Ca+4 ions, but not Mg"1"1" ions (see Figure 8). These experiments used 5μg recombinant tag-free StaOl l, incubated at 37°C for 25 minutes with increasing CaCl2 concentrations from 0.5-5OmM, then analysed by gel electrophoresis on a clear native gel. A mobility shift (indicating oligomerisation) was evident from 2mM Ca+*, and particularly >5mM. These levels compare to blood Ca+* concentrations of about 1.2mM, serum concentrations of about 1 ImM, and milk concentrations of about 32mM. EDTA reversed the shift.
Surface digestion [302] and/or analysis of secreted proteins revealed peptide fragments from CIfA, CIfB, coA, eap, ebhA, ebpS, efb, emp, FnBA, FnBB, hla, IsdA, IsdB, lsdH, ukD, lukS, sdrD, sdrE, sasB, sasD, sasF, spa, staOOl, sta002, staOO3, staOO4, sta005, staOOό, sta007, sta008, sta009, staOlO, staOl l, staO19, staO23, staO24, staO28, staO36, sta040, staO49, staO5O, staO54, staO57, staO64, staO65, staO73, staO95, staO96, staO98, stalOO, stalOl, stalO2, stalO3, stalO5, stalO7, stalO8, stalO9, stal l l, stal l2, stal l3, stal l5, stal lό, stal l7, stal l8, stal20, NW_06, NW_07, NW_08, NW_09 and NW lO e.g. SEQ ID NOs: 228 and 229 were identified as fragments of staO19.
Conjugated capsular saccharides are useful for providing opsonic immunity. Serotypes 5 and 8 cover about 85% of clinical isolates.
Strain coverage
A panel of 36 clinical isolates was used to represent circulating strains, including strains belonging to the five clonal lineages representing the vast majority of worldwide circulating CA-MRSA (community-associated methicillin-resistant S.aureus). HA-MRSA (hospital-associated MRSA) and non-MRSA strains were also included. Overall the panel included 9 HA-MRSA strains, 7 CA-MRSA strains, 2 MRSA strains, and 18 other strains.
Genes encoding IsdA, HIa, EsxA, StaOOό, StaOl l, Spa, and CIfB were present in all 36 strains. The gene for EsxB was absent from 11/36 strains, and the gene for SdrD was absent from 6/36 strains.
The encoded IsdA sequences were 95-100% identical across the panel, and the protein was expressed in iron-limited conditions in the stationary growth phase. The encoded SdrD sequences were 95- 100% identical in the 30/36 SdrD+ve panel members. The encoded EsxA sequences were 100% identical across the panel; the encoded EsxB sequences were 95-100% identical in the 25 EsxB+ve strains. The encoded CIfB sequences were 93-100% identical across the panel, and this protein was also found to be highly surface-exposed in the early exponential growth phase.
Conservation in the encoded amino acid sequences were as follows (% identity):
Figure imgf000124_0001
A larger panel of 61 strains was screened for the presence of genes encoding HIa and StaOOό, as well as for their expression. This panel covered both MRSA and MSSA strains, a variety of geographical origins, and a variety of ST and clonal complex types. 9/61 strains did not express HIa, whereas all but one strain expressed StaOOό (data for the 61st strain were inconclusive). Thus a vaccine based on HIa alone is unlikely to give adequate coverage for a universal vaccine, but this problem could be overcome by addition of StaOOό.
Cytotoxicity and cell binding studies
The analysis of the potential cellular cytotoxicity by S.aureus recombinant antigens HIa, Hla-H35L, IsdA, IsdB, staOOό, staOl l and EsxAB was conducted on HBMECs and A549 cells. Annexin V and propidium iodide staining were used to measure the percentage of early and late apoptotic cells by flow cytometry. Endothelial cells were grown in 24 well plates up to fully confluent. Cells were then incubated for 24 hours with three different concentration of recombinant antigens (lOμg/ml, lμg/ml, O.lμg/ml). The combination of TNF-α and cycloheximide (CHX), which has been reported to induce apoptosis in endothelial cells, was used as a positive control. Incubation with PBS buffer alone was a negative control. Analysis was then performed by FACS.
None of the antigens induced a cytotoxic effect on HBMECs or A549 cells. Indeed, the percentage of live cell population compared to control cells remained essentially constant up to 24 hours of incubation, hi contrast, the combination of TNF-α and CHX induced a 25% increase in the number of apoptotic cells.
HBMECs were also used as an in vitro model for testing the binding of S.aureus recombinant antigens to human endothelial cells. HBMECs were grown up to confluence at 37°C in humidified atmosphere in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum, 10% NuSerum, 2mM glutamine, 1 mM pyruvate, 1% non-essential amino acids, 1% MEM vitamins, 100 units/ml penicillin, and 100 μg/ml streptomycin. Binding of recombinant antigens to the cells was tested by indirect immunofluorescence and analyzed by FACS. The cells positive for binding were measured as net mean intensity of fluorescence respect to negative controls, identified as unspecifϊc antibody recognition. Binding experiments were performed at 4°C. Mouse polyclonal antibodies specific for each of the recombinant antigens were used as primary antibodies and binding was detected by R-Phycoerythrin-conjugated goat anti-mouse IgG secondary antibody. As negative control, HBMECs were incubated with primary polyclonal antibodies detected by fluorescence- labeled secondary antibody or fluorescence-labeled secondary antibody alone. Binding of a known surfaced-exposed GBS antigen was used as positive control. HIa and Hla-H35L were the only antigens able to strongly bind to endothelial cells. The haemolytic activity of these two antigens was also tested.
De-fϊbrinated sheep and rabbit blood were used to measure their haemolytic activity by spectrophotometric assay. The blood was incubated at 37°C for 30 minutes with serial dilution 1 :4 of the two proteins. Incubation with water, to cause osmotic lysis, and incubation with a S.pyogenes protein, were positive controls; as negative control, the blood was incubated with PBS+ BSA 0.5%.
Recombinant native HIa, but not its H35L mutant form, showed haemolytic activity on rabbit erythrocytes. The mutant was at least 150-fold less haemolytic than wild-type. Both proteins had no haemolytic activity on sheep blood.
Thus the S.aureus recombinant vaccine candidates do not show any cytotoxicity both on A549 epithelial cell line and HBMEC endothelial cell line. Importantly, HIa, a secreted toxin known to form pores into the plasma membrane of host cells, could bind A549 cells but did not induce cytotoxicity on them; it was also able to induce haemolysis of rabbit erythrocytes, hi contrast, recombinant Hla-H35L, a variant toxin with a single amino acid substitution that cannot form cytolytic pores, did not induce cellular damage in both human cell lines and rabbit erythrocytes. These findings indicate that this mutant form of HIa may be more safely used in a vaccine composition. None of the other antigens showed the capacity to bind to host cells. Adjuvant formulation
Selected S.aureus protein antigen candidates have been formulated with aluminium hydroxide, either individually or as a combination of proteins, with or without capsular polysaccharide conjugate(s). The formulations have been optimized for pH and osmolality. The antigens were EsxA-B, StaOOό, StaOl l, Hla-H35L, SdrD, IsdA, ISdA40-I84, StaO19, StaO21, StaO73, CIfB45-552, SdrD53-592, SasF, and IsdB. These are formulated as monovalent antigens at lOOμg/ml, or as combinations at 50μg/ml each. Capsular saccharide conjugates from type 5 or type 8 strains are added at 5μg/ml, lOμg/ml or 25μg/ml. Aluminium hydroxide was used at 2mg/ml, in a 1OmM histidine buffer (pH 6.5) and with 9mg/ml NaCl. All monovalent and combination formulations, with or without conjugates, could be adjusted with respect to a desired pH and osmolality. The formulations had pH in the range 6.2-7.3, and osmolality in the range 248-360 mθsm/kg. Glycerol was excluded from formulations as it had a negative impact on osmolality.
All proteins tested, in various monovalent and combination formulations, adsorbed well to the aluminium hydroxide adjuvant, except for IsdA, ISdA40-I84, StaO19, and StaO73.
The individual StaOOό, StaOl 1, EsxA-B and Hla-H35L proteins were completely adsorbed, and could be desorbed without altering their pre-adsorption electrophoretic profile.
Each antigen in a combination of StaOOό, StaOl 1, EsxA-B and Hla-H35L was completely adsorbed, with no inter-antigen competition for the adjuvant. The antigens in a combination of StaOOό, StaOl 1, EsxA-B and ISdA40-184 were also completely adsorbed, except for ISdA40-I84, which behaved in the same way as the monovalent protein. For both combinations, the antigens could be desorbed without altering their pre-adsorption electrophoretic profile.
The additional presence of type 5 and/or type 8 conjugates also did not change the adsorption or desorption characteristics of the antigens e.g. in combination with StaOOό+StaOl 1+EsxA-B. A short stability study (2 weeks at 4°C) was performed to evaluate the stability of monovalent formulations and to evaluate antigen integrity. All tested formulations were stable for their pH and osmolality. All antigens remained completely adsorbed to the adjuvant. All antigens maintained their desorption characteristics. There was no evidence of increased degradation or aggregation of antigens after desorption.
Efficacy testing
Individual antigens staOOό, staOl l, staO12, staO17, staO19, staO21 and staO28 were tested for their ability to protect against IV challenge by 1.2x107 cfu of Newman strain (type 5). Results are shown in Figure 1. All antigens reduced bacterial numbers compared with the control, and the best results were seen with staOOό, staOl 1 and staO19. Further individual antigens were tested: (i) NW lO; (ii) ISdA40-184; (iii) Sta002; (iv) Sta003; (v) Sta073; (vi) StalOl; (vii) StaO14; (viii) HIa-PSGS; (ix) SdrDCnaB- The increase in survival, compared to the negative control group, 15 days after challenge was: (i) 50%; (ii) 19%; (iii) 37%; (iv) 43%; (v) 25%; (vi) 12%; (vii) 25%; (viii) 56%; (ix) 39%. Two hybrid polypeptides were also tested: (i) HlaH35L-EsxAB; (ii) Sta006-EsxAB. The increase in survival after challenge, compared to the negative control group, was: (i) 25%; (ii) 25%.
Table 2 gives a summary of results obtained with various antigens in the abscess model.
Experiment SA-10-a tested the efficacy of antigen combinations. Six groups of twelve CD-I mice received a negative control (PBS), IsdB, or one of the following combinations, adjuvanted with aluminium hydroxide: (i) EsxAB + Hla-H35L; (ii) StaOOό + StaOl 1 + EsxAB; (iii) StaOOό + StaOl 1 + EsxAB + Hla-H35L; or (iv) StaOOό + StaOl 1 + ISdA4O-I84 + EsxAB. Two administrations were given, at days 0 and 14. At day 24 mice received 3x108 cfu of Newman strain staphylococcus and survival in each group was assessed every 24 hours for two weeks. Results are shown in Figure 2. After 14 days, 25% of animals in the positive control group had survived, but 50% of animals in group (ii) had survived, as had 58% of animals in groups (iii) & (iv), and 75% in group (i).
Experiment SA-10-b used the same methods to test: (i) CIfB45-552 + Hla-H35L + StaOOό + EsxAB; (ii) CIfB45-552 + StaOl 1 + StaOOό + EsxAB; (iii) CIfB45-552 + ISdA40-I84 + StaOOό + EsxAB; or (iv) SdrD53-592 + ISdA4O-I84 + StaOOό + EsxAB. Results are shown in Figure 3. After 14 days, 25% of animals in the positive control group and in group (ii) had survived, but 33% of animals in group (iv) had survived, 75% of animals in group (i), and 83% of animals in group (iii).
Further combinations were also used to immunise mice. The combinations were typically adjuvanted with aluminium hydroxide (see above) and were administered on days 0 and 14. The immunisations were in CDl mice, 12 per group. On day 24 the mice were challenged with a lethal dose of live bacteria and survival was then followed for 14 further days. For comparison, PBS was used as a negative control and IsdB as a positive control [2].
Experiment SA-I l tested: (i) a type 5 conjugate combined with EsxAB + StaOOό + StaOl 1; (ii) EsxAB + StaOl 9 + StaOOό + StaOl 1; (iii) a type 5 conjugate + Hla-H35L + StaOOό + StaOl 1; (iv) EsxAB + Hla-H35L + StaOOό + StaOl 1; or (v) EsxAB + ISdA40-184 + StaOOό + StaOl 1. 14 days after challenge all of the negative control animals had died, but 42% of positive control animals had survived. Survival results in the test groups were as follows: (i) 67%; (ii) 42%; (iii) 75%; (iv) 33%; and (v) 25%.
Experiment SA-12 tested: (i) Hla-H35L + ISdA40-I84 + StaOOό + StaOl 1; (ii) Hla-H35L + EsxAB + StaOOό + StaOl 1; (iii) EsxAB + ISdA4O-IS4 + StaOOό + StaOl 1; (iv) EsxAB + IsdA+ StaOOό + StaOl 1. 14 days after challenge 8% of the negative control animals and 17% of positive control animals had survived. Survival results in the test groups were as follows: (i) 50%; (ii) 50%; (iii) 25%; (iv) 33%. Experiment SA- 14 tested: (i) EsxAB + Hla-H35L + StaOOό + StaOl l; (ii) EsxAB + ISdA40-184 + StaOOό + StaOl l; (iii) StaOOό + StaOl l + StaO19 + EsxAB; (iv) StaOOό + StaOl l + StaO19 + Hla-H35L. 14 days after challenge with 5xlO8 CFU of Newman strain, 18% of the negative control animals and 9% of positive control animals had survived; survival results in the test groups were as follows: (i) 58%; (ii) 67%; (iii) 42%; (iv) 83%. Survival numbers over 14 days are shown in Figure 4, showing that all combinations performed better than the two controls on every post-challenge day.
Experiment SA- 17a tested: (i) EsxAB + StaOOό + StaOl l + serotype 5 conjugate + serotype 8 conjugate; (ii) EsxAB + StaO73 + StaOll + serotype 5 conjugate + serotype 8 conjugate; (iii) EsxAB + Hla-H35L + StaOl l + StaO73. Compared to the negative control, the increase in survival 15 days after challenge with Newman strain was: (i) 17%; (ii) 42%; (iii) 34%. The median survival in groups (ii) and (iii) was the full 15 days, and was 12 days in group (i).
Further antigen combination experiments tested: (a) serotype 5 conjugate + serotype 8 conjugate + EsxAB + StaOOό + StaOl l; (b) Sta002+Sta003+Sta021+NW-10; (c) EsxAB+ HlaH35L + StaOOό + StaO19; and (d) EsxAB + Sta006+Sta019. Compared to the negative control, the increase in survival after challenge with Newman strain was: (a) 37%; (b) 36%; (c) 13%. ; and (d) 0%.
Survival data from studies SA-10, SA-11, SA- 12 and SA- 14 were combined to assess the efficacy of two combinations when compared to PBS or IsdB. "Combo- 1" was EsxAB+Hla- H35L+Sta006+Sta011 (with polypeptides comprising SEQ ID NOs: 241, 150, 246 & 247). "Combo-2" was EsxAB+IsdA4o-i84+StaOO6+StaOl l. The median survival times for each group of 48 mice after 14 days were compared. Whereas the PBS and IsdB groups had a median survival time of 1 day, mice in the "Combo- 1" and "Combo-2" groups had a median survival time of 14 days. The differences in median survival duration were compared by a t-test: survival in the "Combo- 1" group was statistically superior to both the PBS group (p<0.0001) and the IsdB group (p<0.0001); survival in the "Combo-2" group was statistically superior to both the PBS group (p<0.0001) and the IsdB group (p=0.0049). These data are shown in Figure 5.
Figure 6 shows data with Combo- 1 and Combo-2 in the abscess model. Kidneys of mice are isolated after challenge and are then homogenised and plated. The cfu count indicates the level of abscess formation. Figure 6 shows data from a single experiment. The numbers beneath the data show the log reduction relative to the PBS group. The reduction is bigger in the two combination groups than with IsdB alone, with U-test (one tail) values of 0.0001 for Combo- 1 and 0.0005 for Combo-2. The same effect was seen in the two combination groups in a second experiment in which an IsdB group was not included.
Further experiments compared protection achieved with Combo- 1, IsdB or PBS against challenge with three different strains: Staph- 19, FPR3757(USA300) and Lac(USA300). There were 44 mice per group and results were as follows (see also Figure 12), including one-tailed p- values for the survival proportion, where: Pl compares Combo- 1 with PBS; P2 compares Combo- 1 with IsdB; and P3 compared PBS with IsdB:
Figure imgf000129_0001
Further experiments showed that immunisation with adjuvanted Combo 1 reduced CFU counts after challenge with Newman, USAlOO, CC30 and US A300 strains, when compared to immunisation with adjuvant alone (aluminium hydroxide) or IsdB. Figure 10 shows CFU values (log/ml) for the four challenge strains. The lowest count, with p<0.015 in each case, was achieved with Combo 1. The area of abscess was also assessed and was also lower in the Combo 1 -immunised mice (e.g. Figure 11).
Further experiments showed that Combo 1 is highly protective against clinically relevant strains in the sepsis model, and always achieved a higher survival % than IsdB. Figure 12 shows that the median survival in Combo 1 -immunised mice (40 per group, 3 experiments) was the full 15 days when challenged with Newman, ST-80, FPR3757 or Lac strains, and that the proportion of mice surviving was >75%. hi contrast, the median survival in IsdB-immunised mice was only 1 day with Newman and ST-80 challenge, with <65% survival for all four challenge strains. Comparison ofCombol to its individual polypeptides
Various tests were performed to compare Combo 1 to its four individual polypeptides (i.e. EsxAB, Hla-H35L, StaOOό, StaOl 1), as well as to IsdB or to an antigen-free negative control.
The opsonophagocytic activity of sera from immunised animals was tested. Sera were obtained using (i) the four individual polypeptides, (ii) all pairs of the polypeptides, (iii) all triplets, or (iv) the full Combo 1 combination. For comparison, anti-IsdB serum was used. Pre- immune and negative control sera showed no killing of Newman strain in this assay. In a first experiment: anti-IsdB serum showed 27% killing; sera against each of the four individual polypeptides showed between 26-34% killing; all multi-polypeptide combinations showed at least 34% killing; and sera raised with Combo-1 showed 39% killing, hi a second experiment sera with Combo-1 showed 43% killing but anti-IsdB serum performed slightly better; all single or multi-polypeptide sera using the Combo-1 polypeptides showed at least 26% killing.
Further experiments looked at passive protection achieved by transferring into mice (20 per group, 8 week old CDl mice) antiserum from immunised rabbits. Four groups received 200μl of sera from rabbits immunised with one of EsxAB, Hla-H35L, StaOOό, StaOl l; a fifth group received 50μl of each serum (200μl in total). Two other groups received serum from IsdB-immunised rabbits or serum from rabbits immunised with saline+adjuvant. 15 minutes later the mice were challenged intraperitoneally (108 CFU of Newman strain) and then mortality was assessed after 14 days. Results were as follows:
Figure imgf000130_0001
In further experiments the level of specific antibodies induced in CDl mice were examined to assess the immunogenicity of the four polypeptides in Combol. Compositions included either 20μg of each of the four single polypeptides, or 4x1 Oμg in the combination. The compositions included an aluminium hydroxide adjuvant. Serum levels of antigen-specific IgG were determined by Luminex 4Plex assay As shown in Figure 9, all four polypeptides were highly immunogenic in CDl mice on their own and in combination. In each case the titer against a polypeptide was higher when it was administered in the combination than when administered alone (compare middle and right pairs).
Further experiments compared protection achieved either with Combo- 1 or with its four individual polypeptides. IsdB was also included for comparison. The proportions of animals surviving (40 animals per group) 15 days after challenge with Newman strain, and the average (median) survival in days, were as follows, including a one-tailed p- value of the surviving proportion in comparison with a PBS+adjuvant negative control:
Figure imgf000130_0002
The murine abscess model was used to compare the four individual polypeptides with the Combol combination. In some experiments mice were immunised with IsdB for comparison. Antigens were adjuvanted with aluminium hydroxide, and adjuvant alone was used as a negative control. Figure 7 shows the numbers of bacteria in animals' kidneys after challenge with four different strains. The lowest average counts were seen for the Combol combination.
Challenge experiments were performed following immunisation with (i) the four individual polypeptides, (ii) all pairs, (iii) all triplets, or (iv) the full Combol combination. IsdB or buffer alone were used for comparison. Survival results from 24 mice per group (3 experiments) after challenge with 5x108 CFU of Newman strain are shown in Figure 13. The median survival for IsdB was only 2 days. The median survival for the individual Combol polypeptides ranged from 1-6 days. Pairs of the polypeptides gave median survival of 2-11 days. Triplets gave median survival of 8-15 days. The full Combo 1 combination gave a median survival of the full 15 days, with 59% of mice surviving this long (c/ only 35% with IsdB).
It will be understood that the invention has been described by way of example only and modifications may be made whilst remaining within the scope and spirit of the invention.
TABLE 1: NOMENCLATURE CROSS-REFERENCE
Figure imgf000131_0001
Figure imgf000132_0001
Figure imgf000133_0001
Figure imgf000134_0001
TABLE 2: ABSCESS MODEL RESULTS SUMMARY
Figure imgf000135_0001
Figure imgf000136_0001
Figure imgf000137_0001
Figure imgf000138_0001
* alum = aluminium hydroxide ** Log reduction in kidney CFU
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Claims

1. An immunogenic composition comprising a combination of antigens, said combination comprising two or more antigens selected from the group consisting of: (1) a staOOό antigen; (2) a staOl 1 antigen; (3) a esxA antigen; (4) a esxB antigen; (5) a hla antigen; (6) a ebpS antigen; (7) a efb antigen; (8) a emp antigen; (9) a esaC antigen; (10) a coA antigen; (11) a eap antigen; (12) a
FnBA antigen; (13) a FnBB antigen; (14) a ebhA antigen; (15) a hlgB antigen; (16) a hlgC antigen; (17) a isdA antigen; (18) a isdB antigen; (19) a isdC antigen; (20) a isdG antigen; (21) a isdH antigen; (22) a isdl antigen; (23) a lukD antigen; (24) a lukE antigen; (25) a lukF antigen; (26) a lukS antigen; (27) a nuc antigen; (28) a sasA antigen; (29) a sasB antigen; (30) a sasC antigen; (31) a sasD antigen; (32) a sasF antigen; (33) a sdrC antigen; (34) a sdrD antigen; (35) a sdrE2 antigen; (36) a spa antigen; (37) a clfA antigen; (38) a clfB antigen; (39) a staOOl antigen; (40) a sta002 antigen; (41) a staOO3 antigen; (42) a sta004 antigen; (43) a staOO5 antigen; (44) a sta007 antigen; (45) a staOO8 antigen; (46) a sta009 antigen; (47) a staOlO antigen; (48) a staO12 antigen; (49) a staO13 antigen; (50) a staO14 antigen; (51) a staO15 antigen; (52) a staO16 antigen; (53) a staO17 antigen; (54) a staO18 antigen; (55) a staO19 antigen; (56) a sta020 antigen; (57) a staO21 antigen; (58) a staO22 antigen; (59) a staO23 antigen; (60) a staO24 antigen; (61) a staO25 antigen; (62) a staO26 antigen; (63) a staO27 antigen; (64) a staO28 antigen; (65) a staO29 antigen; (66) a staO3O antigen; (67) a staO31 antigen; (68) a staO32 antigen; (69) a staO33 antigen; (70) a staO34 antigen; (71) a staO35 antigen; (72) a staO36 antigen; (73) a staO37 antigen; (74) a staO38 antigen; (75) a staO39 antigen; (76) a staO4O antigen; (77) a staO41 antigen; (78) a staO42 antigen;
(79) a staO43 antigen; (80) a staO44 antigen; (81) a staO45 antigen; (82) a staO46 antigen; (83) a staO47 antigen; (84) a staO48 antigen; (85) a staO49 antigen; (86) a staO5O antigen; (87) a staO51 antigen; (88) a staO52 antigen; (89) a staO53 antigen; (90) a staO54 antigen; (91) a staO55 antigen; (92) a staO56 antigen; (93) a staO57 antigen; (94) a staO58 antigen; (95) a staO59 antigen; (96) a sta060 antigen; (97) a staO61 antigen; (98) a staO62 antigen; (99) a staO63 antigen; (100) a staO64 antigen; (101) a staO65 antigen; (102) a staO66 antigen; (103) a staO67 antigen; (104) a staO68 antigen; (105) a staO69 antigen; (106) a sta070 antigen; (107) a staO71 antigen; (108) a staO72 antigen; (109) a staO73 antigen; (110) a staO74 antigen; (111) a staO75 antigen; (112) a staO76 antigen; (113) a staO77 antigen; (114) a staO78 antigen; (115) a staO79 antigen; (116) a staO8O antigen; (117) a staO82 antigen; (118) a staO83 antigen; (119) a staO84 antigen; (120) a staO85 antigen; (121) a staO86 antigen; (122) a staO87 antigen; (123) a staO88 antigen; (124) a staO89 antigen; (125) a sta090 antigen; (126) a staO91 antigen; (127) a staO92 antigen; (128) a staO93 antigen; (129) a staO94 antigen; (130) a staO95 antigen; (131) a staO96 antigen; (132) a staO97 antigen; (133) a staO98 antigen; (134) a staO99 antigen; (135) a stalOO antigen; (136) a stalOl antigen; (137) a stalO2 antigen; (138) a stalO3 antigen; (139) a stalO4 antigen; (140) a stalO5 antigen; (141) a stalO6 antigen; (142) a stalO7 antigen; (143) a stalO8 antigen; (144) a stalO9 antigen; (145) a stal lO antigen; (146) a stal l l antigen; (147) a stal l2 antigen; (148) a stal l3 antigen; (149) a stal l4 antigen; (150) a stal l5 antigen; (151) a stal lό antigen; (152) a stal l7 antigen; (153) a stal l8 antigen; (154) a stal l9 antigen; (155) a stal20 antigen; (156) a NW 6 antigen; (157) a NW_9 antigen; (158) a NW lO antigen; (159) a NW_7 antigen; (160) a NWJ antigen; (161) a NW 2 antigen; (162) a NW l antigen; (163) a staO81 antigen; and (164) a NW 5 antigen.
2. The composition of claim 1, comprising at least one antigen selected from numbers (3) to (38) and at least one antigen selected from numbers (1), (2) and (37) to (149).
3. The composition of claim 2, comprising: at least one antigen selected from numbers (37), (38), (8), (9), (3), (4), (5), (17), (18), (19), (31), (32), (33), (34), (35) and (36); and at least one antigen selected from (40), (1), (43), (2), (64), (96), (133) and (147).
4. The composition of claim 1, comprising two or more antigens selected from the group consisting of: (1) a clfA antigen; (2) a clfB antigen; (3) a sdrE2 antigen; (4) a sdrC antigen; (5) a SasF antigen; (6) a emp antigen; (7) a sdrD antigen; (8) a spa antigen; (9) a esaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) a sta006 antigen; (13) a isdC antigen; (14) a hla antigen; (15) a staOl 1 antigen; (16) isdA antigen; (17) a isdB antigen; (18) a sasF antigen.
5. The composition of claim 1, two or more antigens selected from the group consisting of: (1) a esxA antigen; (2) a esxB antigen; (3) a sta006 antigen; (4) a hla antigen; (5) a staOl 1 antigen.
6. The composition of any preceding claim, wherein one or more of said antigens is adsorbed to an aluminium hydroxide adjuvant, and optionally wherein the composition includes a histidine buffer.
7. The composition of any preceding claim, further comprising: one or more conjugates of (i) a S.aureus exopolysaccharide and (ii) a carrier protein.
8. The composition of any preceding claim, further comprising: one or more conjugates of (i) a S.aureus capsular polysaccharide and (ii) a carrier protein.
9. A polypeptide of formula NH2-A- {-X-L-}π-B-COOH, wherein:
X is an amino acid sequence of a staphylococcal antigen, selected from the group consisting of S.aureus antigens sta006, staOl 1, esxA, esxB, hla, clfA, clfB, coA, eap, ebhA, ebpS, efb, emp, esaC, FnBA, FnBB, hlgB, hlgC, isdA, isdB, isdC, isdG, isdH, isdl, lukD, lukE, lukF, lukS, nuc, sasA, sasB, sasC, sasD, sasF, sdrC, sdrD, sdrE2, spa, staOOl, sta002, sta003, sta004, staOO5, sta007, sta008, sta009, staOlO, staO12, staO13, staO14, staO15, staOlό, staO17, staO18, staO19, sta020, staO21, staO22, staO23, staO24, staO25, staO26, staO27, staO28, staO29, staO3O, staO31, staO32, staO33, staO34, staO35, staO36, staO37, staO38, staO39, sta040, staO41, staO42, staO43, staO44, staO45, staO46, staO47, staO48, staO49, staO5O, staO51, staO52, staO53, staO54, staO55, staO56, staO57, staO58, staO59, sta060, staO61, staO62, staO63, staO64, staO65, staO66, staO67, staO68, staO69, sta070, staO71, staO72, staO73, staO74, staO75, staO76, staO77, staO78, staO79, staO8O, staO81, staO82, staO83, staO84, staO85, staO86, staO87, staO88, staO89, sta090, staO91, staO92, staO93, staO94, staO95, staO96, staO97, staO98, staO99, stalOO, stalOl, stalO2, stalO3, stalO4, stalO5, stalO6, stalO7, stalO8, stalO9, stal lO, stal l l, stal l2, stal l3, stal l4, stal l5, stal 16, stal 17, stal 18, NW_6, NW_9, NW_10, NW_7, NWJ, NW_2, NW_1, and NWJ; L is an optional linker amino acid sequence; A is an optional N-terminal amino acid sequence; B is an optional C-terminal amino acid sequence; and n is an integer of 2 or more.
10. An immunogenic composition comprising the polypeptide of claim 9 and further comprising:
(A) one or more conjugates of (i) a S.aureus exopolysaccharide and (ii) a carrier protein; and/or
(B) one or more conjugates of (i) a S.aureus capsular polysaccharide and (ii) a carrier protein.
11. The composition or polypeptide of any preceding claim, wherein the clfA antigen can elicit an antibody which recognises SEQ ID NO: 1 and comprises an amino acid sequence: (a) having 80% or more identity to SEQ ID NO: 1; and/or (b) comprising a fragment of at least 7 consecutive amino acids of SEQ ID NO: 1, wherein the fragment comprises an epitope from SEQ ID NO: 1.
12. A polypeptide comprising amino acid sequence (a) having 80% or more identity to SEQ ID NO: 151; and/or (b) comprising a fragment of at least 7 consecutive amino acids from amino acids 1-97 of SEQ ID NO: 151 and at least 7 consecutive amino acids from amino acids 104-207 of
SEQ ID NO: 151, wherein the polypeptide can elicit antibodies which recognise both the wild- type staphylococcal protein comprising SEQ ID NO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO: 11.
13. An immunogenic composition comprising the polypeptide of claim 12 and one or more of (i) a staOOό antigen; (ii) a hla antigen; and/or (iii) a staOl 1 antigen.
14. The composition of claim 13, including an adjuvant.
15. A polypeptide comprising amino acid sequence having 80% or more identity to an amino acid sequence selected from SEQ ID NOs: 151, 152, 168, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 220, 221, 222, 223, 224, 237, 238, 241.
16. A polypeptide comprising an amino acid sequence selected from SEQ ID NOs: 151, 152, 168, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 220, 221, 222, 223, 224, 237, 238, 241, 242, 243, 244, 245.
17. A polypeptide comprising: (a) a first sequence having 90% or more identity to SEQ ID NO: 218; and (b) a second sequence having 90% or more identity to SEQ ID NO: 219, wherein the first and second sequences are either directly joined or are joined by an intervening amino acid sequence having fewer than 10 amino acids.
18. A pharmaceutical composition comprising the polypeptide of any one of claims 12, 15, 16 or 17.
19. A method for raising an immune response in a mammal comprising the step of administering to the mammal an effective amount of the polypeptide or composition of any preceding claim.
20. Nucleic acid encoding the polypeptide of claim 9, 12, 15, 16 or 17.
PCT/IB2010/000998 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aureus WO2010119343A2 (en)

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NZ595689A NZ595689A (en) 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aureus
CN2010800274105A CN102647999A (en) 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aerus
SG2011072832A SG175092A1 (en) 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aerus
EP10723299A EP2419129A2 (en) 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aerus
RU2011146016/15A RU2508126C2 (en) 2009-04-14 2010-04-14 Compositions for immunisation against staphylococcus aureus
JP2012505252A JP5830009B2 (en) 2009-04-14 2010-04-14 Composition for immunization against Staphylococcus aureus
CN201810736674.6A CN109248313B (en) 2009-04-14 2010-04-14 Compositions for immunization against staphylococcus aureus
US13/264,369 US8679505B2 (en) 2009-04-14 2010-04-14 Compositions for immunising against Staphylococcus aureus
BRPI1013780A BRPI1013780B8 (en) 2009-04-14 2010-04-14 IMMUNOGENIC COMPOSITION USEFUL FOR IMMUNIZATION AGAINST STAPHYLOCOCCUS AUREUS, ITS PREPARATION METHOD AND PHARMACEUTICAL COMPOSITION
MX2015006258A MX363222B (en) 2009-04-14 2010-04-14 Compositions for immunising against staphylococcus aerus.
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AU2010238255A AU2010238255B2 (en) 2009-04-14 2010-04-14 Compositions for immunising against Staphylococcus aureus
US13/234,077 US8632783B2 (en) 2009-04-14 2011-09-15 Compositions for immunising against Staphylococcus aureus
IL215593A IL215593A0 (en) 2009-04-14 2011-10-06 Compositions for immunising against staphylococcus aureus
US14/171,537 US9205142B2 (en) 2009-04-14 2014-02-03 Compositions for immunising against staphylococcus aureus
PH12015500092A PH12015500092A1 (en) 2009-04-14 2015-01-14 Compositions for immunising against staphylococcus aureus
US14/961,772 US9642904B2 (en) 2009-04-14 2015-12-07 Compositions for immunising against Staphylococcus aureus
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