WO1986001808A1 - Anticorps monoclonaux et leur utilisation - Google Patents

Anticorps monoclonaux et leur utilisation Download PDF

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Publication number
WO1986001808A1
WO1986001808A1 PCT/GB1985/000410 GB8500410W WO8601808A1 WO 1986001808 A1 WO1986001808 A1 WO 1986001808A1 GB 8500410 W GB8500410 W GB 8500410W WO 8601808 A1 WO8601808 A1 WO 8601808A1
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WIPO (PCT)
Prior art keywords
monoclonal antibody
campylobacter
antigen
immunoassay
enzyme
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PCT/GB1985/000410
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English (en)
Inventor
Bruce William Wright
Peter John Cox
Alice Margaret Noyes
Danny Widdows
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Technology Licence Company Limited
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Publication date
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Publication of WO1986001808A1 publication Critical patent/WO1986001808A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
    • C07K16/121—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Helicobacter (Campylobacter) (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)

Definitions

  • MONOCLONAL ANTIBODIES AND THEIR USE This invention relates to monoclonal antibodies and their use.
  • Campylobacter infections • are primarily of importance in the field of veterinary medicine. Serious economic losses to farmers result from abortions and infertility of infected cattle and sheep. Infection of animals is transmitted venereally and may be harboured asymptomatically for long periods of time in the genito-urinary and intestinal tracts.
  • C_. fetus infections can be made only by culture of the organism. The organisms grow on various media, but careful attention must be given to the atmospheric conditions. There is little or no growth under strict anaerobic conditions. . fetus is microaerophilic; in a semi-solid medium, growth occurs only within the first few millimeters below the surface. This is in contrast to some other Campylobacter that will grow under conditions of strict anaerobiosis.
  • Antibodies in high titers may be demonstrated in patients with documented infection, by the use of an indirect he agglutination test. Experience with this test, however, is still limited. Campylobacter species associated with human disease are frequently resistant to penicillin, cephalothin and carbenicillin. Most strains which have been tested are sensitive to gentamicin, chloramphenicol and tetracycline. Recurrences have been reported following short courses of therapy.
  • Campylobacter The members of the genera Campylobacter are the agents of a specifically human enteric disease, bacterial dysentery. Bacterial diarrhea is a common and often serious condition manifest as fluid loss from the bowel, leading in many cases to dehydration, and occasionally death.
  • Campylobacter In addition to diarrhea, Campylobacter is known to cause gram-negative sepsis which is a bloodstream infection. It is one of the major infectious disease problems encountered in modern medical centres. While it can be transient and self-limited, severe gram-negative sepsis constitutes a medical emergency.
  • Campylobacter infections vary depending on the locus of the infection. It is estimated that in the United States and Europe many millions of cases of bacterial diarrhea occur annually, of which several million are seen by a physician or admitted to a hospital. Because of the self-limiting nature of the adult disease, most people do not seek treatment. Of the people seeking treatment, bacterial diagnosis of diarrhea is presently made by stool culture techniques. These techniques are generally performed only in hospitals and are slow, requiring one to three days. During this time, the patient is exposed, if treated, to the expense and potential hazard of inappropriate therapy. However, if not treated, the patient is exposed to the hazard of a deteriorating condition pending the test result and initiation of therapy.
  • test for gram-negative sepsis involves processing blood and urine cultures and other procedures on occasion.
  • blood culture tests are cumbersome. They require a day, and often several days, to return results. They require expert laboratory skills because of the complex nature of human blood which tends to interact non-specifically with many of the test reagents.
  • the present invention provides novel monoclonal antibodies for use in accurately and rapidly diagnosing samples for the presence of Campylobacter antigens and/or organisms.
  • the present invention comprises monoclonal antibodies specific for an antigen of Campylobactery in particular, the antigens of Campylobacter fetus, including the sub-species (intestinalis jejuni) thereof, the antigens of Campylobacter sputorum, including the sub-species (bubulus, mucosalis) thereof, and the antigens of Campylobacter fecalis, as well as a monoclonal antibody broadly cross-reactive with an antigen for each species (or substantially all species) of the genus Campylobacter.
  • the invention also comprises labelled monoclonal antibodies for use in diagnosing the presence of the Campylobacter antigens, each comprising a monoclonal antibody against one of the above-mentioned antigens to Campylobacter or to a particular species thereof and having linked thereto an appropriate label.
  • the label can be, for example, a radioactive isotope, enzyme, fluorescent compound, chemiluminescent compound, bioluminescent compound, ferromagnetic atom or particle.
  • the invention further comprises the process for diagnosing the presence of Campylobacter antigens or organisms in a specimen, comprising contacting said specimen with the labelled monoclonal antibody in an appropriate immunoassay procedure.
  • the invention is also directed to a therapeutic composition
  • a therapeutic composition comprising a monoclonal antibody for an antigen of Campylobacter and a carrier or diluent, as well as kits containing at least one labelled monoclonal antibody to an antigen of a Campylobacter.
  • the monoclonal antibodies of the present invention are prepared by fusing spleen cells from a mammal which has been immunised against the particular Campylobacter antigen, with an appropriate myeloma cell line, preferably NSO (uncloned) , P3NS1-Ag4/1, or Sp2/0 Agl4. The resultant product is then cultured in a standard HAT (hypoxanthine, aminopterin and thymidine) medium. Screening tests for the specific monoclonal antibodies are employed utilising immunoassay techniques which will be described below.
  • the immunised spleen cells may be derived from any mammal, such as primates, humans, rodents (i.e.
  • mice, rats and rabbits mice, bovines, ovines and canines, but the present invention will be described in connection with mice.
  • the mouse is first immunised by injection of the particular Campylobacter antigen chosen, e.g. for a period of approximately eleven weeks. When the mouse shows sufficient antibody production against the antigen, as determined by conventional assay, it is given a booster injection of the appropriate Campylobacter antigen, and then killed so that the immunised spleen may be removed. The fusion can then be carried out utilising immunised spleen cells and an appropriate myeloma cell line.
  • the fused cells yielding an antibody which gives a positive response to the presence of the particular
  • Campylobacter antigen are removed and cloned utilising any of the standard methods.
  • the monoclonal .antibodies from the clones are then tested against standard antigens to determine their specificity for the particular Campylobacter antigen.
  • the monoclonal antibody selected, which is specific for the particular Campylobacter antigen or species, is then bound to an appropriate label.
  • Amounts of antibody sufficient for labelling and subsequent commercial production are produced by the known techniques, such as by batch or continuous tissue culture or culture ii vivo in mammals such as mice.
  • the monoclonal antibodies may be labelled with various labels, as exemplified above.
  • the present invention will be described with reference to the use of an enzyme-labelled monoclonal antibody. Examples of enzymes utilised as labels are alkaline phosphatase, glucose oxidase, galactosidase, peroxidase and urease.
  • Such linkage with enzymes can be accomplished by any known method, such as the Staphylococcal Protein A ethod, the glutaraldehyde method, the benzoquinone method, or the periodate method.
  • EIA enzyme-linked immunosorbent assay
  • a non-labelled antigen and a specific antibody are combined with identical fluorescently-labelled antigen. .Both labelled and unlabelled antigen compete for antibody binding sites. The amount of labelled antigen bound to the antibody is dependent upon, and therefore a measurement of, the concentration of non-labelled antigen.
  • Examples of this particular type of fluorescent-immunoassay include heterogeneous systems such as Enzyme-Linked Fluorescent Immunoassay, or homogeneous systems such as the Substrate-Labelled Fluorescent Immunoassay.
  • the most suitable fluorescent probe, and the one most widely used, is fluorescein. While fluorescein can be subject to ' considerable interference from scattering, sensitivity can be increased by the use of a fluorometer optimised for the probe utilised in the particular assay, and in which the effect of scattering can be minimised.
  • Fluorescence polarisation In fluorescence polarisation, a labelled sample is excited with polarised light and the degree of polarisation of the emitted light is measured. As the antigen binds to the antibody, its rotation slows down and the degree of polarisation increases. Fluorescence polarisation is simple, quick and precise. However, at the present time, its sensitivity is limited to the micromole per litre range and upper nanomole per litre range with respect to antigens in biological samples.
  • Luminescence is the emission of light by an atom or molecule as an electron is transferred to the ground state from a higher energy state.
  • the free energy of a chemical reaction provides the energy required to produce an intermediate reaction or product in an electronically-excited state. Subsequent decay back to the ground state is accompanied by emission of light.
  • Bioluminescence is .the name given to a special form of chemiluminescence found in biological systems, in which a catalytic protein or enzyme, such as luciferase, increases the efficiency of the luminescent reaction. The best known chemiluminescent substance is luminol.
  • a further aspect of the present invention is a therapeutic composition
  • a therapeutic composition comprising one or more of the monoclonal antibodies to the particular Campylobacter antigen or species, as well as a pharmacologically-acceptable carrier or diluent.
  • Such compositions can be used to treat humans and/or animals afflicted with some form of shigellosis and they are used in amounts effective to cure; the amount may vary widely, depending upon the individual being treated and the severity of the infection.
  • One or more of the monoclonal antibodies can be assembled into a diagnostic kit for use in diagnosing for the presence of an antigen, antigens or species of Campylobacter in various specimens. It is also possible to use the broadly cross-reactive monoclonal antibody which can identify the genus Campylobacter alone or as part of a kit containing antibodies that can identify other bacterial genera or species of Campylobacter and/or other bacteria. in the past, there have been difficulties in developing rapid kits because of undesirable cross-reactions of specimens; e.g. faeces with antiserum. The use of monoclonal antibodies can eliminate these problems and provide highly specific and rapid tests for diagnosis.
  • conjugated or labelled monoclonal antibodies for antigens and/or species of Campylobacter and other gram-negative bacteria can be utilised in a kit to identify such antigens and organisms in blood samples taken from patients for the diagnosis of possible Campylobacter or other gram-negative sepsis.
  • the monoclonal test is an advance over existing procedures in that it is more accurate than existing tests; it gives "same day” results, provides convenience to the patient and improves therapy as a result of early, accurate diagnosis; and it reduces labour costs and laboratory time required for administration of the tests.
  • the kit may be sold individually or included as a component in a comprehensive line of compatible immunoassay reagents sold to reference laboratories to detect the species and serotypes of Campylobacter.
  • One preferred embodiment of the present invention is diagnostic kit comprising at least one labelled monoclonal antibody against a particular Campylobacter antigen or species, as well as any appropriate stains, counterstains or reagents. Further embodiments include kits containing at least one control sample of a Campylobacter antigen and/or a cross-reactive labelled monoclonal antibody which would detect the presence of any of the given particular Campylobacter organisms in a particular sample.
  • Monoclonal diagnostics which detect the presence of Campylobacter antigens can also be used in periodic testing of water sources, food supplies.and food processing operations.
  • the present invention describes the use of the labelled monoclonal antibodies to determine the presence of a standard antigen
  • the invention can have many applications in diagnosing the presence of antigens by determining whether specimens, such as urine, blood, stool, water ' and milk, contain the particular Campylobacter antigen. More particularly, the invention could be utilised as a public health and safety diagnostic aid, whereby specimens such as water or food could be tested for possible contamination.
  • the monoclonal antibodies of the present invention were prepared generally according to the method of Kohler and Milstein, supra.
  • API Analytical Profile Index (ref. Ayerst Laboratories)
  • DMEM Dulbeccos Modified Eagles Medium
  • FCS Foetal Calf Serum
  • % T refers to vaccine concentrations measured in a 1 cm light path
  • PBS Phosphate Buffered Saline
  • CFA Complete Freunds
  • Adjuvant ip intraperitoneal
  • Campylobacter fetus was obtained from the National Collection of Type Cultures (NCTC accession No. 10842) and tested by standard biochemical methods of microbial identification to confirm its identity (using API profiles) .
  • the Campylobacter fetus was removed from the lyophile, grown on blood agar, and tested by API to confirm its identity and purity.
  • the bacteria were transferred for growth on to blood agar (microaerophilic) and harvested. The organisms were washed in saline by repeated centrifugation, and were then resuspended iri for ol saline.
  • mice were injected with the prepared antigen, over five weeks. They were given one intraperitoneal injection per week for three weeks (0.1 ml 80% T vaccine) , followed by an intravenous injection each week for three weeks of Campylobacter fetus vaccine prepared as above. The mice were bled approximately six days after the last injection and the serum tested for antibodies by assay. The conventional assay used for this serum titer testing was the enzyme-linked immunosorbent assay system. When the mice showed antibody production after this regimen, generally a positive titer of at least 10,000, a mouse was selected as a fusion donor and given a booster injection (0.05 ml 80% T vaccine) intravenously, three days prior to splenectomy.
  • the myeloma cell line used was NS0 (uncloned) , obtained from the MRC Laboratory of Molecular Biology in Cambridge, England. The myeloma cells were in the log growth phase, and rapidly dividing ' . Each cell line was washed using, as tissue culture medium, DMEM containing 3% FCS. The spleen cells were then spun down at the same time that a relevant volume of myeloma cells were spun down (room temperature for 7 minutes at 600 g) , and each resultant pellet was then separately resuspended in 10 ml 3% FCS-DMEM.
  • 0.1 ml • of the suspension was diluted to 1 ml and a haemacytometer with phase microscope was used.
  • 0.1 ml of the suspension was diluted to 1 ml with Methyl Violet-citric acid solution, and a haemacytometer and light microscop'e were used to count the stained nuclei of the cells.
  • the resultant cell pellet was placed in a 37°C water-bath. 1 ml of a 50 w/v solution of polyethylene glycol 1500 (PEG) in saline Hepes, pH approximately 7.5, was added, and the mixture gently stirred for approximately 1.5 minutes. 10 ml serum-free tissue culture medium DMEM were then slowly added, followed by up to 50 ml of such culture medium, centrifugation and removal of all the supernatant, and resuspension of the cell pellet in 10 ml of DMEM containing 18% by weight
  • dilutions of cells suspensions in 18% FCS-DMEM + Balb/c mouse macrophages were made to achieve 1 cell/well and half cell/well in a 96-well microtitre plate.
  • the plates were incubated for 7-14 days at 37 C, 95% RH, 7-9% C0 2 until semi-confluent.
  • the supernatants were then assayed for specific antibody by the standard enzyme immunosorbent assay.
  • the monoclonal antibodies from the clones were screened by the standard techniques for binding to Campylobacter fetus NCTC 10842, prepared as in the immunisation, and for specificity in a test battery of Campylobacter fetus and related species bearing different antigens. Specifically, a grid of microtitre plates containing a representative selective of organisms was prepared, boiled, and utilised as a template to define the specificity of the parent group. The EIA immunoassy noted above may be used.
  • the monoclonal showed the appropriate specificity * (to NCTC 10842) . It was negative with respect to other Campylobacter, IS. coli, Serratia, Providencia, Proteus, Haemophilus, Enterobacter, Klebsiella, Pseudomonas, Shigella and Salmonella.
  • mice were primed with pristane for at least 7
  • Purification was accomplished using the protein A - Sepharose method. Ascites fluid was filtered through glass wool and centrifuged at 30,000 g for 10 minutes.
  • the ascites was then diluted with twice its own volume of cold phosphate buffer (0.1M sodium phosphate, pH 8.2).
  • the benzoquinone method was used.
  • alkaline phosphatase (Sigma Type VII-T) were dialysed against 2 x 500 ml of 0.25 M sodium phosphate buffer, pH 6.0, at +4 C. 18 mg p-benzoquinone were dissolved in 0.6 ml warm AR ethanol, and added to the dialysed alkaline phosphatase. The benzoquinone/alkaline phosphatase mixture was left in the dark at room temperature for 1 hour. Unreacted benzoquinone and reaction by-products were then removed and the buffer exchanged by gel filtration on a Pharmacia PD-10
  • the conjugate was dialysed against 2 x 1000 ml PBS + 0.02% sodium azide at +4 C. An equal volume of glycerol was added. The conjugate was sterile-filtered through a 0.22 ⁇ m membrane filter into a sterile amber vial, and stored at +4 C.
  • the enzyme immunoassay method was used for testing. This method comprises coating the wells of a standard polyvinyl chloride (PVC) microtitre tray with the antigen, followed by addition of monoclonal antibody enzyme conjugate, and finally addition of the enzyme substrate, para-nitrophenyl phosphate.
  • PVC polyvinyl chloride
  • the monoclonal antibodies were found to be specific for the antigen of Campylobacter fetus.
  • the monoclonal antibody was also tested and shown to be of the Class IgG2a.
  • the particular epitopic site to which the antibody attaches to the antigen can also be determined.
  • the same enzyme immunoassay method can also be used to determine whether diagnostic specimens -such as urine, blood, stool, water or milk contain the antigen. In such cases, the antibody can first be bound to the plate.
  • Example 1 The procedure of Example 1 was followed in each of six cases, with differences outlined below, to prepare monoclonal antibodies and conjugates for various antigens of the genus Campylobacter.
  • Example 2 the antigen was Campylobacter fetus venerialis NCTC 10842; in Example 3, Campylobacter fetus subsp. fetus, NCTC 10348; and in Examples 4 to 7, Campylobacter jejuni Penner types 2 (NCTC 11168) , 6 (NCTC 11392) , 23 (NCTC 11351) and 9 (NCTC 11349) , respectively.
  • the organisms were washed in formol saline.
  • Examples 4 to 7 after the use of blood agar, the organisms were harvested and resuspended in formol saline. Microaero- philicity was observed in Examples 3, 4, 5, 6 and 7.
  • Example 2 In the animal immunisation step, the sequences of injections and intervals (in weeks) were: in Example 2, ip-2-ip-l-ip-5-iv-l-iv-l-iv-6-iv; in Example 3, im (in
  • the dilution method was used in addition, in Example 2.
  • dilutions of cells suspensions in 18% FCS-DMEM + Balb/c mouse macrophages were made to achieve 1 cell/well and half cell/well in a 96-well microtitre plate.
  • the plates were incubated for 7-14 days at 37 C, 95% RH, 7-9% C0 2 until semi-confluent.
  • the supernatants were then assayed for specific antibody by the standard enzyme immunosorbent assay.
  • Example 2 Additionally in Example 2, and as the antibody production procedure in Examples 3 and 6, cells of the monoclonal antibody-producing cell line were grown in batch tissue culture. DMEM-10% FCS was used to support growth in mid-log phase, to 1 litre volume. The culture was then allowed to overgrow, to allow maximum antibody production. The culture was then centrifuged at 1200 g for approximately 10 minutes, the cells discarded and the antibody-rich supernatant collected.
  • Antibody purification for Examples 2 and 3 was conducted as follows:
  • TRIS buffered supernatant was applied at a flow rate of 1 ml/min to a 1 ml column of Protein A-Sepharose, previously equilibrated with 0.1M TRIS buffer, pH 8.2. The column was then washed with 40 ml of 0.1M TRIS buffer.
  • the monoclonal antibody was eluted with citrate buffer (0.1M sodium citrate, pH 3.5) into sufficient IM TRIS buffer, pH 9.0, to raise the pH immediately to .about 7.5.
  • the eluate was dialysed in PBS, pH 7.4, at 4 C, and stored at -20 C. Two procedures were used for antibody purification in Example 6.
  • ascites fluid was filtered through glass wool and centrifuged at 30,000 g for 10 minutes. The ascites was then stirred at +4 C and an equal volume of cold, saturated ammonium sulphate added slowly. The mixture was stirred for a further 30 minutes after the addition was complete. The precipitate was harvested by centrifugation at 10,000 g for 10 minutes. The precipitate was dissolved in a minimum volume of cold phosphate/EDTA buffer (20 mM sodium phosphate, 10 mM EDTA, pH 7.5, + 0.02% sodium azide) . The solution was dialysed vs 2 x 1000 ml o'f the same buffer at +4 C.
  • the dialysed, redissolved precipitate was centrifuged at 30,000 g for 10 minutes and applied to a 10 ml column of DEAE-cellulose, previously equilibrated in phosphate/EDTA buffer.
  • the monoclonal antibody was eluted with phosphate/EDTA buffer.
  • the suspension was stirred for a further 30 minutes, and then the precipitate was harvested by centrifugation at 10,000 g for 10 minutes.
  • the precipitate was dissolved in a minimum volume of cold phosphate/EDTA buffer (20 mM sodium phosphate, 10 mM EDTA pH 7.5 + 0.02% sodium azide) .
  • the dialysed, redissolved precipitate was centrifuged at 30,000 g for 10 minutes and applied to a 10 ml column of DEAE-cellulose, previously equilibrated in phosphate/EDTA buffer.
  • the monoclonal antibody was eluted with phosphate/EDTA buffer.
  • the monoclonals showed the appropriate specificity, and were negative with respect to other Campylobacter.
  • the monoclonal of Example 2 was specific to NCTC 10842 and NCTC 10348, both bearing the antigen of Campylobacter fetus venerialis.
  • the monoclonals of Examples 2, 5 and 6 were negative with respect to E. coli, Shigella and Salmonella, the monoclonals of Examples 2 and 5 were negative to ' Enterobacter, the monoclonal of Example 2 was negative to Proteus, Providencia and Serratia, the monoclonal of Example 5 was negative to Klebsiella, and the monoclonal of Example 6 was negative to Pseudomonas.
  • Example 2 The monoclonal antibodies obtained in Examples 2, 3, 5 and 6 were respectively of the Sub-classes IgG3, IgG3, IgG2b and IgGl.
  • Example 8 The procedure of Example 1 was repeated, to give a monoclonal antibody broadly cross-reactive with all species of the genus Campylobacter.
  • Tests using the present invention are superior to existing tests, based on the following advantages: (i) greater accuracy; (ii) same day results, within an hour or two; (iii) reduction in amount of skilled labour required to administer laboratory procedures, resulting in reduced labour costs; (iv) reduction in laboratory time and space used in connection with tests, resulting in reduced overhead expenses; and (v) improved therapy based upon early, precise diagnosis. While the invention has been described in connection with certain preferred embodiments, it is not intended to limit the scope of the invention to the particular form set forth but, on the contrary, it is intended to cover such alternatives, modifications, and equivalents as may be included withiii the spirit and scope of the invention as defined by the appended claims.

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Abstract

Des anticorps monoclonaux pour le genre Campylobacter, les anticorps marqués, compositions et kits les contenant, et leur utilisation pour le diagnostic d'antigènes et le traitement.
PCT/GB1985/000410 1984-09-07 1985-09-09 Anticorps monoclonaux et leur utilisation WO1986001808A1 (fr)

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GB848422651A GB8422651D0 (en) 1984-09-07 1984-09-07 Monoclonal antibodies
GB8422651 1984-09-07

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987001119A1 (fr) * 1985-08-16 1987-02-26 Gregory Murray Winn Anticorps monoclonaux reactifs contre le campylobacter pyloridis
EP0329570A2 (fr) * 1988-02-18 1989-08-23 Martin J. Blaser Compositions antigéniques composées de fragments de Campylobacter pylori et procédés pour leur production et utilisation
WO1990003575A1 (fr) * 1988-09-29 1990-04-05 Public Health Laboratory Service Board PROCEDE PERMETTANT DE PRODUIRE DES ANTIGENES SPECIFIQUES $i(CAMPYLOBACTER PYLORI)
US6057115A (en) * 1995-06-16 2000-05-02 Ludwig Institute For Cancer Research Process for producing GM2 specific antibodies
US9221898B2 (en) 2010-02-11 2015-12-29 The Governors Of The University Of Alberta N-linked glycan compounds

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0077734A2 (fr) * 1981-10-19 1983-04-27 Mgi Pharma, Inc. Production d'anticorps monoclonaux contre des adhésines bactériennes
WO1983001739A1 (fr) * 1981-11-17 1983-05-26 Brigham & Womens Hospital Anticorps monoclonaux contre le brugia malayi
EP0101039A2 (fr) * 1982-08-10 1984-02-22 Meiji Seika Kabushiki Kaisha Anticorps monoclonal, méthode pour sa production et son utilisation
EP0105714A1 (fr) * 1982-09-29 1984-04-18 Serono Diagnostics Limited Immunoessai pour antigènes
EP0111762A1 (fr) * 1980-06-20 1984-06-27 Unilever Plc Procédés et appareil pour l'exécution d'essais de liaisons spécifiques

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0111762A1 (fr) * 1980-06-20 1984-06-27 Unilever Plc Procédés et appareil pour l'exécution d'essais de liaisons spécifiques
EP0077734A2 (fr) * 1981-10-19 1983-04-27 Mgi Pharma, Inc. Production d'anticorps monoclonaux contre des adhésines bactériennes
WO1983001739A1 (fr) * 1981-11-17 1983-05-26 Brigham & Womens Hospital Anticorps monoclonaux contre le brugia malayi
EP0101039A2 (fr) * 1982-08-10 1984-02-22 Meiji Seika Kabushiki Kaisha Anticorps monoclonal, méthode pour sa production et son utilisation
EP0105714A1 (fr) * 1982-09-29 1984-04-18 Serono Diagnostics Limited Immunoessai pour antigènes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Biological Abstracts, Volume 77, No. 11, 1984, Philadelphia, PA, (US) T.U. KOSUNEN et al.: "Analysis of Campylobacter Jejuni Antigens with Monoclonal Antibodies", see page 9222, column 2, Abstract Nr. 83483 & J. Clin. Microbiol., 1984, 19(27), 129-133 (Eng) *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987001119A1 (fr) * 1985-08-16 1987-02-26 Gregory Murray Winn Anticorps monoclonaux reactifs contre le campylobacter pyloridis
EP0329570A2 (fr) * 1988-02-18 1989-08-23 Martin J. Blaser Compositions antigéniques composées de fragments de Campylobacter pylori et procédés pour leur production et utilisation
EP0329570A3 (fr) * 1988-02-18 1991-05-22 Martin J. Blaser Compositions antigéniques composées de fragments de Campylobacter pylori et procédés pour leur production et utilisation
US5459041A (en) * 1988-02-18 1995-10-17 Enteric Research Laboratories, Inc. Campylobacter pylori antigens and uses thereof for detection of Campylobacter pylori infection
WO1990003575A1 (fr) * 1988-09-29 1990-04-05 Public Health Laboratory Service Board PROCEDE PERMETTANT DE PRODUIRE DES ANTIGENES SPECIFIQUES $i(CAMPYLOBACTER PYLORI)
US6057115A (en) * 1995-06-16 2000-05-02 Ludwig Institute For Cancer Research Process for producing GM2 specific antibodies
US9221898B2 (en) 2010-02-11 2015-12-29 The Governors Of The University Of Alberta N-linked glycan compounds

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GB8422651D0 (en) 1984-10-10
EP0192728A1 (fr) 1986-09-03
JPS62500170A (ja) 1987-01-22

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