WO2024083847A1 - Antibodies and antibody derivatives against equine il-5 - Google Patents

Antibodies and antibody derivatives against equine il-5 Download PDF

Info

Publication number
WO2024083847A1
WO2024083847A1 PCT/EP2023/078860 EP2023078860W WO2024083847A1 WO 2024083847 A1 WO2024083847 A1 WO 2024083847A1 EP 2023078860 W EP2023078860 W EP 2023078860W WO 2024083847 A1 WO2024083847 A1 WO 2024083847A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
acid sequence
amino acid
seq
equine
Prior art date
Application number
PCT/EP2023/078860
Other languages
French (fr)
Inventor
Nora Langreder
Dorina SCHÄCKERMANN
Michael Hust
Kristine ROSSBACH
Simone LADEL
Original Assignee
Wirtschaftsgenossenschaft Deutscher Tierärzte Eg
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wirtschaftsgenossenschaft Deutscher Tierärzte Eg filed Critical Wirtschaftsgenossenschaft Deutscher Tierärzte Eg
Publication of WO2024083847A1 publication Critical patent/WO2024083847A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C07K16/244—Interleukins [IL]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • 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
    • 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
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • Allergic conditions affect about 10% of horses (including horses, ponies, donkeys, hinnies and mules) worldwide.
  • seasonal allergies are the most common and they mainly belong to the group of skin allergies and respiratory allergies.
  • the most prominent examples of skin and respiratory allergies are insect-bite hypersensitivity and asthma.
  • IBH Insect-bite hypersensitivity
  • CH Culicoides hypersensitivity
  • sweet itch summer eczema
  • kasen kasen
  • Queensland itch summer dermatitis or equine dermatitis
  • CH mosquitos and midges
  • This most common allergic dermatitis may affect up to 72% of allergic horses (including ponies) depending on race and region, especially in the warmer regions, and is a reaction of the animal to the saliva of midges (Culicoides) and other insects.
  • the bites lead to an allergic reaction with a massive infiltration of eosinophil cells to the allergic skin area.
  • horses suffer of pruritus, skin lesions, hair loss and secondary infections, which can lead to chronic dermatitis. They experience itchiness, which is associated with the insect bite (pruritus), and this can lead to cases, where the animals scratch that strong so they hurt themselves leading to open skin lesions with alopecia and crusting. These lesions may then promote secondary infections. Thus, horses severely affected from summer eczema cannot live a normal herd life. Horses with asthma suffer from chronical airway inflammation that causes a poor performance to a cough, nasal discharge or overt respiratory distress.
  • Asthma includes conditions ranging from mild asthma, also called inflammatory airway disease (IAD) in young horses, to severe asthma, widely referred to as heaves (chronic obstructive pulmonary disease (COPD) or recurrent airway obstruction (RAO)) and pasture- or summer-associated airway disease.
  • IAD inflammatory airway disease
  • COPD chronic obstructive pulmonary disease
  • REO recurrent airway obstruction
  • the clinical signs primarily develop after exposure to aerosolized particles, which could be endotoxins produced by specific bacteria, particles originating from molds, fungi, beta-D-glucan, microorganisms, vegetative material, inorganic dusts, and noxious gases, such as ammonia from urine, mite debris and other.
  • Severely asthmatic horses are usually managed through antigen avoidance and the use of corticosteroids and bronchodilators to reduce airway inflammation, bronchoconstriction and improve lung function (Simoes, J. Equine Vet. Sci.2020, 87, 102937). However, some horses are unresponsive to corticosteroid treatment and an effective treatment is still needed. Like with other allergic conditions, also in asthma eosinophilic airway inflammation is observed. Interleukin-5 (IL-5) is connected to various chronic inflammatory conditions and seems to be involved in allergic conditions, including equine allergic asthma and insect-bite hypersensitivity, which is the most common allergic condition in horses affecting the skin (Janssen et al..
  • equine IBH shows a type I allergy phenotype, including allergen-derived CD4 type-2 T helper (Th2) polarization and IL-4/IL-13 predominance. This is explained to lead to Culicoides allergen-specific IgE and corresponding degranulation of mast cells and basophiles.
  • Eosinophilia has been suggested to play an increasingly dominant role in chronic allergic disease phases, whereas early phases are dominated by Type I-IgE mediated reactions (Fettelunter- Gabriel et al.. The Veterinary Journal 2021,276, 105741).
  • Th2 cTh2
  • peTh2 pathogenic effector Th2
  • IL-5 is the most potent activator ofeosinophils and is produced by Th2 cells and ILC2s in mammals.
  • IL-5 is also reported to have a role in eosinophil extracellular trap cell death (Nagase et al., Allergology International 2020, 69, 178-186). It binds to the interleukin-5 receptor and is, therefore, involved in B-cell growth and immunoglobulin secretion. As a key mediator in eosinophil activation, IL-5 was identified by the inventors as a target for addressing allergic conditions, such as IBH, atopic dermatitis and asthma, in horses. The important role of eosinophils in the molecular mechanism ofatopic dermatitis in horses can be seen e.g., in Fettelberger-Gabriel et al. (2021).
  • IL-5 was also shown to be upregulated in other allergic horses and, in particular, in horses with asthma (Janssen et al.. Frontiers in Immunology 2022, 12, 921077). Dewachi et al. (2006) found out that horses with heaves had significantly increased numbers ofneutrophils expressing IL-5 receptors compared to control while in pasture, and further increased during stabling in heaves affected horses but not in control animals (Dewachi et al., Vet Immunol Immunopathol 2006, 109(1-2), 31-6). Using immunohistochemistry and in situ hybridization, the expression ofIL-5 was observed in the BALF lymphocytes of sEA-affected horses (Cordeau et al., Vet. Immunol.
  • VLP virus-like particle
  • equine IL-5 vaccine for the treatment ofIBH.
  • the vaccine has been tested in horses with IBH in placebo-controlled randomized double-blind clinical trials and both showed reduction of IBH lesion scores after vaccination throughout the whole IBH season.
  • these virus-based approaches have the disadvantage that the immune reaction may be triggered excessively due to the polyclonal glycosylated antibodies, which are produced in the horse's body after vaccination. Further there might be the risk of the generation of autoimmune antibodies for life-time as an endogenous IL-5 protein is used for vaccination. Fettelberger-Gabriel et al. (2021) explicitly teaches away from the use of therapeutic antibodies in horses.
  • At least partially means a part but, maybe, not the whole entity to which the term is referring to. By indicating “at least”, everything between a part and the whole entity is comprised.
  • the backbone typically comprises the constant parts, i.e., the constant region of the heavy and of the light chain, of an antibody.
  • "at least partially equine” with respect to the equine backbone comprises the full constant regions of the heavy and/or light chain of the antibody as well as parts thereof, like tmncated forms.
  • parts of the backbone may be derived from an organism other than horse. It is, however, preferred that the whole backbone is equine, i.e., is of equine origin. More preferably, the backbone is of equine origin but glycosylation sites are removed by amino acid exchange or chemical modification so that no glycosyl residue may be attached.
  • the above-described diagnostic antibodies of the prior art are mainly derived from goat or rabbit and may not be used as a medicament since they would trigger an immune response from the horse's immune system against the antibodies of the foreign species.
  • the prior art antibodies are not at least partially equine and do not comprise at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof.
  • the diagnostic antibodies also comprise the naturally occurring glycosylation sites.
  • an efficient treatment of horses suffering from allergic conditions is not available to date.
  • the inventors have developed antibodies and antibody derivatives specifically binding to IL-5 in horses. Those antibodies show high binding activity and high neutralization efficiency in in vitro studies. Further, they have also been able to show a high safety and tolerability in target animal studies that support the effectiveness of the antibodies in the treatment of allergic conditions in horse, in particular, in the treatment of IBH.
  • the present invention provides an isolated antibody or antibody derivative that specifically binds to equine IL-5 and comprises at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof.
  • the presence of at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof diminishes the activation of the horse's immune system against the administered antibodies or antibody derivatives. Thereby, the effectiveness of the antibody or antibody derivative therapy is increased.
  • the present invention explicitly and preferably provides aglycosylated antibodies and antibody derivatives, which, in contrast to the VLP-based vaccine, explicitly avoid triggering the immune system of the horse, i.e., does not lead to a further enhancement of the immime reaction of the horse.
  • the IgG subclass more preferably, the IgG6 subclass, may be used. This subclass is known to not interact with cells of the innate and adaptive immune system. Thereby, an over-reaction of the horse to the antibody or antibody derivative of the invention as well as the generation of autoimmune reactions is prevented.
  • antibodies and antibody derivatives of the IgG and IgG6 only have a blood half-life of about 21 days.
  • the basic vaccination regimen with the VLP-based vaccine consists of three vaccinations in the first year and an annual booster for the following years. The was higher in the second treatment and paralleled by more sustained antibody. Vaccinated horses had no IL-5-specific antibodies shortly before the booster in the second and the third year, indicating the reversibility of vaccine-induced anti-IL-5 antibodies. However, whether repeated vaccination over a period of years may eventually permanently change the immune system cannot be inferred from the study available so far.
  • the advantage of monoclonal antibodies is that they do not influence the immune system itself, but directly capture IL-5 (Fettelunter-Gabriel et al..
  • the present invention provides an isolated antibody or antibody derivative that specifically binds to equine IL-5 comprising at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof.
  • the isolated antibody or antibody derivative comprises at least one equine constant light chain or fragments thereof and/or at least one equine constant heavy chain or fragments thereof, optionally, as depicted in SEQ ID NO: 56 (constant region light chain amino acid sequence), SEQ ID NO: 57 (constant region heavy chain amino acid sequence) or SEQ NO: 151 (amino acid sequence CH1 constant region of heavy chain), respectively, or having at least 75%, at least 80%, at least 90%, preferably, at least 95% or 97%, more preferably 98% and, particularly preferred, 99% sequence homology thereto.
  • the isolated antibody or antibody derivative comprises at least two equine light chain(s) and at least two equine constant heavy chain(s) or fragments thereof, optionally as depicted in SEQ ID NO: 56 (constant region light chain amino acid sequence), SEQ ID NO: 57 (constant region heavy chain amino acid sequence) or SEQ ID NO: 151 (amino acid sequence CH1 constant region of heavy chain), respectively, or having at least 75%, at least 80%, at least 90%, preferably, at least or 97%, more preferably 98% and, particularly preferred, 99% sequence homology thereto.
  • the isolated antibody or antibody derivative is a monoclonal antibody or antibody derivative.
  • the (monoclonal) isolated antibody or antibody derivative is aglycosylated.
  • the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 1,155, 156,157,158,2,3, 4, 5, 6, 7, 8, 9, 10, 11,12,13, 14, 15, 16,17,18, 19, 20 and 21.
  • CDR complementary determining region
  • the isolated antibody or antibody derivative comprises at least one CDR amino acid sequence having at least 99% sequence homo logy to any of the sequences as depicted in any one ofSEQ ID NOs: 1,155,156,157,158,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and21.
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155,156, 157 and 158, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3, 4 and 5, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6, 7, 8 and 9, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 10, 11, 12, 13 and 14, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQIDNOs: 15, 16, 17, 18, 19, 20 and 21.
  • the isolated antibody or antibody derivative according to the invention comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23, 24, 25, 26, 27, 28,29,30,31, 32, 33, 35, 36 or 37, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39, 40,41,42,43,45, 49,50,51,52 or 53.
  • the isolated antibody or antibody derivative according to the invention comprises a light chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 54, and/or comprises a heavy chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 55 or 153, more preferably as in SEQ ID NO: 55.
  • the isolated antibody or antibody derivative is an isolated IgG antibody or antibody derivative. More preferably, the isolated antibody or antibody derivative is an isolated IgG6 antibody or antibody derivative.
  • the isolated antibody or antibody derivative according to the invention has a neutralizing ability towards IL-5, in particular, equine IL-5.
  • the antibody does not bind to bovine serum albumin (BSA) or only to a very minor extent.
  • BSA bovine serum albumin
  • the isolated antibody or antibody derivative according to the invention is equinized.
  • the present invention also provides an isolated nucleic acid molecule comprising at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention.
  • a vector comprising at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention.
  • the vector is an expression vector.
  • a host cell comprising the vector of the invention or at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention and as described herein.
  • the host cell is a eukaryotic host cell.
  • the invention provides a pharmaceutical composition comprising the isolated antibody or antibody derivative according to the invention and a pharmaceutically acceptable excipient.
  • the isolated antibody or antibody derivative or the pharmaceutical composition according to the invention and as described herein are for use as a medicament.
  • the isolated antibody or antibody derivative or the pharmaceutical composition of the invention are provided for use in the treatment of an allergic condition of a horse.
  • the allergic condition is selected from a group consisting of equine allergic skin disease, equine allergic respiratory disease and inflammatory disease.
  • the equine allergic skin disease is selected from a group consisting of IBH, atopic dermatitis, food hypersensitivity, allergic and irritant contact dermatitis and urticaria, more preferably, the equine allergic skin disease is IBH or atopic dermatitis, most preferably, the equine allergic skin disease is IBH.
  • the equine allergic respiratory disease is, preferably, equine asthma. It is also preferred that the allergic condition is IBH or equine asthma.
  • an in vitro method for the diagnosis of an allergic condition in a horse comprising contacting the isolated antibody or antibody derivative according to the invention with a biological sample, preferably obtained from a horse, which might be suspected of suffering from an allergic condition.
  • a method of manufacturing the isolated antibody or antibody derivative according to the invention comprising expressing the isolated antibody or antibody derivative in a host cell.
  • the isolated antibody or antibody derivative is expressed in a host cell according to the invention, such as a eukaryotic cell.
  • antibody denotes an intact immunoglobulin having two light and two heavy chains.
  • An isolated antibody or antibody derivative may be a polyclonal antibody, a monoclonal antibody, a synthetic or artificial antibody, a recombinant antibody, a bi-, tri- or multi-specific antibody, a chimeric antibody and/or an equinized antibody. Also combinations thereof are possible, e.g., the antibody may be a monoclonal and equinized antibody.
  • An "anti-IL5" or "a-IL5" antibody is an antibody, which binds to IL-5, in particular to equine IL-5, as the antibody of the present invention. Preferably, the antibody of the invention specifically binds to equine IL-5.
  • the antibody of the present invention is further defined in the claims.
  • IgA immunoglobulin A
  • IgD immunoglobulin D
  • IgE immunoglobulin E
  • IgG immunoglobulin G
  • IgM immunoglobulin M
  • the heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 8, s, y, and [i, respectively.
  • the subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known in multiple species. The prevalence of individual isotypes and functional activities associated with these constant domains are species- specific.
  • the "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (K) and lambda (X), based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes.
  • An ,intact immunoglobulin” or “native immunoglobulin” of the IgG subclass is usually a heterotetrameric glycoprotein having a molecular weight of about 150,000 Daltons, composed of two identical light (L) chains and two identical heavy chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH, heavy chain variable region) followed by a number of constant domains (heavy chain region).
  • VH variable domain
  • constant domains heavy chain region
  • Each light chain has a variable domain at one end (VL, light chain variable region) and a constant domain at its other end (light chain constant region); the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain.
  • VL variable domain
  • region constant region particular amino acid residues are believed to form an interface between the light and heavy chain variable domains (regions).
  • the variable domains (regions) of the isolated antibody and antibody derivative of the present invention bind to the antigen, i.e., to equine IL-5.
  • the isolated antibody or antibody derivative of the present invention is, preferably, aglycosylated.
  • antibody derivative includes all short or abbreviated forms of the antibody described herein as well as antibody fragments, i.e., to non-intact immunoglobulins.
  • An antibody derivative includes the antigen binding portion of the antibody, which binds to, and preferably neutralizes, specifically equine IL-5.
  • the antibody derivative may be truncated in the Fc region of the antibody or may not contain an Fc region at all.
  • the term includes isolated single (heavy or light) antibody chains, an Fv construct, F(ab')2 fragment, a Fab construct, an Fc construct and a light or heavy chain variable or complementarity determining region (CDR) sequence.
  • antibody derivative comprises antibody conjugates, wherein the antibody is conjugated to another functional molecular entity, optionally via a linker, such as a short peptide (1-10 amino acids) or another organic linker.
  • Conjugated denotes a covalent bond between the antibody and another molecular entity, optionally, an organic or inorganic molecular moiety, which may equip the antibody with additional properties and/or functionalities. This may include at least a moiety to facilitate purification, detection, increase bioavailability and/or biocompatibility.
  • a molecular entity may also be used to localize the antibody and antibody derivative to a desired region within the horse's body.
  • the linker may be cleavable by chemical means or by applying heat, light or other.
  • a fragment of equine constant light chain or equine constant heavy chain includes any truncated, i.e., shortened, form of an equine constant light chain or equine constant heavy chain and may contain only few (at least 10 amino acids).
  • the term "hypervariable region”, as used herein, refers to the amino acid residues of an antibody or antibody derivative, which are responsible for antigen binding, e.g., bind to equine IL-5.
  • the hypervariable region comprises amino acid residues from a "complementarity determining region" (CDR) and/or those residues from a "hypervariable loop".
  • “Framework” (FR) residues are those variable domain residues other than the hypervariable region residues.
  • Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab” (Fragment antigen binding) fragments, each with a single antigen-binding site, and a residual "Fc” fragment, named after its ability to crystallize.
  • Pepsin treatment of antibodies yields an F(ab')2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.
  • Fv is the minimum antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. It is in this configuration that the three hypervariable regions of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer.
  • the six hypervariable regions confer antigen-binding specificity to the antibody.
  • a single variable domain or half of an Fv comprising only three hypervariable regions specific for an antigen
  • the Fab also contains the constant domain of the light chain and the first constant domain of the heavy chain (CH1).
  • Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain including one or more cysteine(s) from the antibody hinge region.
  • Fab'-SH is the designation for Fab ⁇ in which the cysteine residue(s) of the constant domains bear a thiol group.
  • F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments, which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.
  • the term "specifically”, in the context of antibody or antibody derivative binding, refers to high avidity and/or high binding activity of an antibody/derivative to a antigen, i.e., a polypeptide, or epitope.
  • the binding activity of a protein- protein interaction can be described as the strength of the interaction between a receptor and its respective ligand.
  • An example of this is an antibody's or antibody derivative's epitope and an antigen (here, in particular, IL-5).
  • the antigen binds to the epitope through a variety of different types of bonds, such as hydrogen bonds, ionic bonds, van der Waals interactions and electrostatic forces.
  • the term “specifically” also includes that the antibody or antibody derivative binds to a far greater extent to equine IL-5 than to other targets, including BSA.
  • isolated means that the antibody, antibody derivative or nucleic acid molecule is separated and/or recovered from a component of a natural environment. Contaminant components of a natural environment are materials that would interfere with diagnostic or therapeutic uses for the material, and may include enzymes, and other proteinaceous solutes.
  • an isolated nucleic acid may include one that is separated from the 5' to 3' sequences, with which it is normally associated in the chromosome.
  • the material will be purified to greater than 95% by weight of the material, and most preferably more than 99% by weight.
  • Isolated material includes the material in situ within recombinant cells since at least one component of the material's natural environment will not be present. Ordinarily, however, isolated material will be prepared by at least one purification step.
  • IL-5" interleukine-5 is the most potent activator of eosinophils and is produced by Th2 cells and ILC2s in mammals.
  • IL-5 is also reported to have a role in eosinophil extracellular trap cell death and binds to the IL-5 receptor. It is further involved in B- cell growth and immunoglobulin secretion. Moreover, IL-5 was also shown to be upregulated in other allergic horses and, in particular, in horses with asthma.
  • the antibody and antibody derivative of the present invention specifically binds to an equine IL-5, i.e., an IL-5 from horse.
  • a "monoclonal antibody or antibody derivative” as defined herein is a single antibody or antibody derivative that is generated by hybridoma technology or that is selected by a phage display approach and cloned into an immunoglobulin, preferably equine IgG backbone (i.e., constant regions are equine and belong to the IgG or IgG6 immunoglobulin class). This construct may then be transferred on a DNA basis into single-cloned eukaryotic cells for production.
  • a “chimeric” antibody is an antibody, in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or artificially created, as well as fragments of such antibodies, as long as they exhibit the desired biological activity.
  • chimeric antibodies are antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from antibody variable and constant region genes belonging to different species or are artificially designed. For example, the variable segments of the genes from a non-equine monoclonal antibody may be joined to equine constant segments.
  • variable domains including the antigen binding site is derived from a human antibody library while the constant domains are of equine origin.
  • the isolated antibody or antibody derivative of the present invention is an isolated chimeric antibody or antibody derivative, preferably, an isolated chimeric antibody or antibody derivative comprising at least one member selected from a group consisting of a non-equine light chain variable region, non- heavy chain variable region, at least one non-equine CDR and combinations thereof.
  • “None-equine” includes all organisms except horses, i.e., animals, which belong to the family ofEquidae. Examples of none-equine is human.
  • “Equine” in the sense of the present invention is to be understood as “derived originating from horse” or “of horse origin”.
  • the antibodies and antibodies derivatives of the present invention also bind to IL-5 from donkeys, hinnies and mules.
  • "Homologous” or having a certain percentage of sequence homology with respect to amino acid or nucleic acid sequences means that the sequences or antibody chains share at least 80% sequence identity, preferably at least 85% or at least 90%, more preferably at least 95% or 97% and even more preferably at least 98% or 99% sequence or homology. Most preferred is a sequence identity of 100%. In this case, the comprised amino acid or nucleic acid sequence is 100% identical to the respective SEQ ID NO.
  • sequence identity or homology with respect to an amino acid or nucleic acid sequence is defined herein as the percentage ofamino acid or nucleic acid residues in the differing sequence that are identical with the parent or original amino acid or nucleic acid residues, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. None of N-terminal, C-terminal, or internal extensions, deletions, or insertions into the antibody sequence shall be construed as affecting sequence identity or homology.
  • sequence identity or homology may be determined by standard computational tools available to the skilled person, such as BLAST or similar.
  • Equinized forms ofnon-equine (e.g., marine) antibodies are genetically engineered antibodies that contain partly sequences derived from non-equine immunoglobulin.
  • Equinized antibodies are equine immunoglobulin sequences (recipient antibody) in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-equine species (donor antibody), such as human or artificial, having the desired specificity, binding activity and capacity.
  • donor antibody such as human or artificial
  • framework region (FR) residues of the equine immunoglobulin sequences are replaced by corresponding non-equine residues.
  • equinized antibodies include residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance.
  • the equinized antibody will include substantially all or at least one, and typically two, variable domains, in which all or substantially all of the hypervariable regions correspond to those of a non-equine immunoglobulin sequence and all or all of the FRs are those of an equine immunoglobulin sequence.
  • the equinized antibody also will comprise a complete or at least a portion of an immunoglobulin constant region (Fc), typically that of an equine immunoglobulin sequence.
  • Fc immunoglobulin constant region
  • neutralizing ability towards IL-5" means that the antibody or antibody reaches the IC5o value at a molar ratio of antibody to monomeric antigen in the range of 0.3-1.
  • the IC50 value which is the antibody or antibody derivative concentration needed to reduce the binding to 50% of equine IL-5 (eqIL-5) to its receptor, when preincubated with the antigen and then added to Expi293F cells which present the eqIL-5 receptor on their surface. Binding of the antigen to the cells is detected in the flow cytometer by fluorescent labelling via the His-tag and a fluorescently labelled antibody. To obtain the relative binding, the measured fluorescence signal (median) is to be related to the signal when only antigen and cells, but no inhibiting antibody, were present.
  • glycosylated denotes that the antibody or antibody derivative is not glycosylated due to a mutation in the constant region of the heavy chain, when compared to the native equine constant region heavy chain amino acid sequence. While aglycosylated antibodies or antibody derivatives are equal in terms of antigen binding, pharmacokinetics, and biodistribution, they have the advantage to bypass the drawbacks of glycosylated antibodies that include glycan heterogeneity and requirement of high capital investment for biomanufacturing. For example, glycosylated antibodies or glycosylated antibody derivatives may have an unpleasant role in recmiting innate immimity effector cells.
  • nucleic acid refers to a series of nucleotide bases (also denoted as “nucleotides”) in DNA and RNA.
  • the nucleic acid molecule of the invention may contain deoxyribonucleotides, ribonucleotides, and/or their natural or artificial analogs.
  • nucleic acid molecule includes single-stranded and double- molecules.
  • a nucleic acid molecule can be a gene or gene fragment, exons, introns, a DNA molecule, like cDNA, an RNA molecule, like mRNA, recombinant nucleic acid molecules, plasmids, and other vectors, primers and probes. Both 5' to 3' (sense) and 3' to 5' (antisense) polynucleotides are included.
  • the nucleic acid molecule of the present invention is preferably isolated as defined hereinabove.
  • nucleic acid sequence may encode the full amino acid sequence of the heavy and/or light chain or may encode only parts of the amino acid sequences of the heavy and/or light chain of the antibody or antibody derivatives of the invention, i.e., only the heavy chain or only the light chain, only the variable region of the heavy or light chain or only the constant region of the heavy or light chain.
  • Different combinations are possible and also only parts of the heavy and light chains may be encoded by a single nucleic acid sequence or single nucleic acid molecule.
  • different host cells, (isolated) nucleic acid molecules or vectors may be used.
  • the heavy and light chains of the antibody and antibody derivative may be produced separately in any order or in parallel using different host cells, isolated nucleic acid molecules and/or vectors.
  • antibody derivatives not the full amino acid sequence of the heavy and/or light chain may be needed and the nucleic acid sequences disclosed herein may be used in truncated form, i.e., not in the full length as described herein,to produce e.g., truncated antibody derivatives.
  • the amino acid and nucleic acid sequences described herein may be truncated.
  • host cell refers to a prokaryotic or eukaryotic cell (e.g., bacterial cells or mammalian cells) whether located in vitro or in vivo.
  • host cells may be located in a transgenic animal.
  • Host cell can be used as a recipient for vectors and may any transformable organism that is capable of replicating a vector and/or expressing a heterologous nucleic acid encoded by a vector.
  • the antibody or antibody derivative of the present invention may be produced in large quantities and in reproducible quality.
  • protein includes the antibody and antibody derivative of the present invention.
  • the (expression) vector carrying the nucleic acid sequence encoding the antibody or antibody derivative according to the present invention can be introduced into an appropriate host cell by any of a variety of suitable means, including transformation, transfection, conjugation, protoplast fusion, calcium phosphate-precipitation and application with polycations such as diethylaminoethyl (DEAE) dextran, and such mechanical means as electroporation, direct microinjection, and microprojectile bombardment.
  • the term "allergic condition” is defined herein as a disorder or disease caused by an interaction between the immune system and a substance foreign to the body, i.e., the allergen.
  • the allergic condition is IBH or equine asthma.
  • the allergic condition is IBH.
  • the term includes allergic skin diseases, allergic respiratory diseases and inflammatory diseases.
  • the term "equine allergic skin disease” includes diseases that manifest with symptoms correlated to the skin of the horse, such as IBH, atopic dermatitis, food hypersensitivity, allergic and irritant contact dermatitis and urticaria, preferably, the equine allergic skin disease is IBH or atopic dermatitis, most preferably, the equine allergic skin disease is IBH. Also combinations thereof are possible.
  • IBH insect-bite hypersensitivity
  • CH Culicoides hypersensitivity
  • sweet itch "kasen”
  • Quanteensland itch "summer dermatitis”
  • equine dermatitis and “summer eczema” denote allergic reactions to bites from mosquitos and midges (Culicoides) but also to other insects (percutaneously). These terms may be used interchangeably herein.
  • the disease is characterized by pruritus and secondary lesions ofalopecia and crusting. The distribution on the horse's body depends on the species of Culicoides feeding on the horse.
  • the classic distribution is the dorsally distributed disease with lesions found on the face, mane, withers, rump, and tail (mainly denoted as sweet itch). Ventrally feeding Culicoides cause lesions in the intermandibular space and on the ventral body wall. Combinations of both are possible, too.
  • minimizing the exposure to the insects is the only option for horse owners.
  • glucocorticoids with many adverse effects are the only therapy available but not suitable as durable therapy. Further, it is helpful to stable the affected horses from dusk till dawn as well as to use insect repellents, fans in the stalls, fly masks, and fly sheets.
  • Atopic dermatitis is a pmritic inflammatory disease of the skin in mammals, mainly caused by contact with diverse antigens (e.g., pollen, mold, food, mites and dust, both inhaled and percutaneously) that can be followed by an inflammatory respiratory disease later in disease progress (Misery and Stander, edt.2010. Pruritus. London, Springer, Marsella, VetSci.2021, 8(7), 124.).
  • Clinical signs of equine atopic dermatitis are pruritus and skin inflammation that typically affects the face, ears, and glabrous areas. Many atopic horses are also insect allergic.
  • Food Hypersensitivity is all allergic condition caused by food allergens, like grains, grasses, supplements or food additives.
  • Allergic and irritant contact dermatitis including allergic eczema is an allergic condition that affects the skin of the horse and other mammals and results in a disrupted epidermis associated with an itchy inflammatory response due to allergic sensitization. Allergens include lotions, shampoos etc., nickel, copper, zinc (percutaneously).
  • Allergic contact dermatitis can either occur as a single event or chronically (Nedorost, Dermatologic Clinics 2020, J5(3) 301-308).
  • "Urticaria” (including “hives”) is a skin condition characterized by the formation of itchy red or whitish raised patches, usually caused by an allergen (Misery and Stander Pruritus 2010, London, Springer).
  • An "equine allergic respiratory disease” is an allergic condition of the horse, which manifests in symptoms of the respiratory system of the horse.
  • An "inflammatory disease” is a condition characterized by inflammation.
  • Equine asthma includes all respiratory diseases from inflammatory airway diseases (IAD) with medium to high severity, heaves and Recurrent Airway Obstructions (RAO).
  • IAD inflammatory airway diseases
  • RAO Recurrent Airway Obstructions
  • COB chronic obstruction pulmonary disease
  • COB chronic obstmctive bronchitis
  • a “pharmaceutical composition” in the sense of the invention denotes a combination of active agent, here the antibody or antibody derivative of the invention, and another compound or composition, which can be inert (like water or buffer), or active, such as an adjuvant.
  • a “therapeutically effective amount” refers to an amount of an active ingredient, i.e., the antibody or antibody derivative according to the invention, sufficient to effect beneficial or desired results, when administered to a subject or patient, which is in the present case, the horse affected by the allergic condition.
  • An effective amount can be administered by one or more routes of administration, applications or dosages.
  • Administration may be weekly, every second or third week, once per month, every second, third or six month or yearly depending on the decision of the veterinary based on the disease state and condition of the affected horse. Administration may by any route of administration (see below).
  • a therapeutically effective amount of a composition according to the invention may be readily determined by one of ordinary skill in the art or the horse owner if trained by the veterinary.
  • a "therapeutically effective amount" is one that produces an objectively measurable change in one or more parameters associated with treatment of the allergic condition, including clinical improvement in symptoms or the prevention of worsening of symptoms.
  • the therapeutically effective amount will vary depending upon the particular horse/subject and condition being treated, the race, gender, weight and age of the horse, the severity of the disease condition, the time point of the year in case of seasonal allergies, the particular compound and composition chosen, stabling of the horse, potential additional medicaments taken by the horse, the feeding, the dosing regimen to be followed and the timing of administration.
  • “Treatment”, “treating”, and the like refers to both therapeutic treatment and prophylactic or preventative measures.
  • Animals in need of treatment include those already with the allergic condition as well as those, in which the allergic condition is to be prevented, such as horses, which are suspected to have a predisposition to develop an allergic condition due to, e.g., pedigree or race.
  • treatment or “treating” of an allergic condition includes preventing or protecting against the allergic condition (that is causing the clinical symptoms not to develop); inhibiting the allergic condition (i.e., arresting or suppressing the development of clinical symptoms, and/or relieving the allergic condition (i.e., causing the regression of clinical symptoms).
  • the term “prophylaxis” will be understood to constitute a type of “treatment” that encompasses both "preventing” and “suppressing”.
  • treatment thus, may include "prophylaxis”.
  • the antibodies and antibody derivatives according to the invention provide for a specific diagnostic tool to target IL-5 activity in horses, which can be used as a marker for allergic conditions.
  • diagnostic tools with high sensitivity and specificity need to be developed.
  • the answer to this need is presented by the invention, which provides an isolated antibody or antibody derivative that specifically binds to an equine IL-5.
  • the isolated antibody or antibody derivative are monoclonal, chimeric and/or equinized.
  • the antibodies and antibody derivatives of the present invention may be produced by standard means in standard expression hosts using standard expression vectors.
  • Typical expression hosts comprise bacterial cells or eukaryotic cells, preferably the expression cell or host cell is a eukaryotic cell. Cloning techniques are also well known to the skilled person.
  • the pharmaceutical composition provided with the present invention comprise the antibody or antibody derivative of the present invention and a pharmaceutically acceptable excipient.
  • the antibody or antibody derivative is to be administered in a therapeutically effective amount.
  • the amount or concentration within the pharmaceutical composition depends on the stability that can provided by the pharmaceutical composition to the antibody or antibody derivative. The stability must be suitable enough to ensure a storage for a convenient period of time,;". e., degradation must be in a regulatory accepted range.
  • Pharmaceutically acceptable excipients suitable for use in the present invention are well known to those of skill in the art.
  • Such carriers include, without limitation, water, saline, in particular, normal saline (i.e., a solution of 0.90% w/v ofNaCl), buffered saline, phosphate buffer, phosphate buffered saline, alcoholic and aqueous solutions, emulsions or suspensions.
  • saline normal saline
  • buffered saline phosphate buffer
  • phosphate buffered saline alcoholic and aqueous solutions
  • emulsions or suspensions emulsions or suspensions.
  • Other conventionally employed diluents, adjuvants and excipients may be added in accordance with conventional techniques.
  • Such carriers can include ethanol, polyols, and suitable mixtures thereof, vegetable oils and injectable organic esters. Buffers and pH adjusting agents may also be employed. Buffers include salts prepared from an organic acid or base.
  • Representative buffers are organic acid salts, such as salts of citric acid, like citrates, ascorbic acid, gluconic acid, histidine-HCl, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid, (hydroxymethyl)aminomethane (Tris) or phosphate buffers.
  • Parenteral carriers can include sodium chloride solution, Ringer's dextrose, dextrose, trehalose, sucrose, and sodium chloride, lactated Ringer's or fixed oils.
  • Intravenous carriers comprise fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose and the like.
  • Preservatives and other additives such as, for example, antioxidants, chelating agents (e.g., EDTA), inert gases and the like may also be provided in the pharmaceutical carriers.
  • the preparation of these pharmaceutically acceptable compositions, from the above-described components, having appropriate pH isotonicity, stability and other conventional characteristics is within the skill of the art.
  • the pharmaceutically acceptable excipient is phosphate buffered saline (PBS).
  • PBS phosphate buffered saline
  • concentration of the antibody in the pharmaceutical composition is typically in the range of 0.1 to 20 mg/mL, more preferably in the range of 2 to 10 mg/mL and even more preferably in the range of 5 to 10 mg/mL.
  • compositions according to the invention may be administered via any possible route of administration, such as via the parenteral, oral, subcutaneous, transdermal route, inhalational or pulmonary route, nasal, sublingual/buccal, mucosal, topical, rectal and/or intravenous route.
  • the pharmaceutical composition of the present invention is administered via the subcutaneous route of administration.
  • the present invention also provides for an in vitro method for the diagnosis of an allergic condition in a horse. Detecting IL-5 in companion animals known to be or suspected of having a pruritic and/or allergic condition.
  • a biological, preferably liquid, sample obtained from a horse which may be a fluid obtainable from the body, including a blood or serum sample, tissue sample or else, the presence ofIL-5 may be determined.
  • a reference sample e.g., obtained from a non-allergic horse
  • an increased level of IL-5 may be determined.
  • Contacting may be performed by mixing.
  • An increased level or concentration of IL-5 may then be indicative of an allergic condition of a horse, optionally in combination with a further examination by a veterinary.
  • Methods for detecting and comparing antibody or antibody derivative concentrations in a sample include enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunospot (ELISPOT), Western blot (WB), immunoprecipitation (IP) and others and are well-known to the person of ordinary skill in the art. Information can e.g., be found in "Molekularbiologische Methoden 2.0" (2018, utb, 2nd ed., Thomas Reinard). Particularly useful are immunoassays like ELISA, which detect or quantitate IL-5 or anti-IL-5 antibodies in a sample obtained from a horse.
  • the antibody or antibody derivative of the present invention may be covalently or non-covalently bound to a detectable molecular entity (such as a polypeptide or small molecule), which includes a fluorescent, radioactive or otherwise detectible molecular entity.
  • a detectable molecular entity such as a polypeptide or small molecule
  • An immunoassay for IL-5 typically comprises incubating a clinical or biological sample of the horse in the presence of a detectably labeled antibody or antibody derivative of the present invention capable of selectively binding to IL-5, and detecting the antibody or antibody derivative, which is bound in a sample by standard means depending on the used detectable molecular entity.
  • the biological sample may be any body fluid, including serum, plasma, tissue and cell lysate, saliva, sputum and bronchoalveolar lavage.
  • SEQ ID NO: 1 Light chain CDR 1 (NOL1-13): NIGSKS.
  • SEQ ID NO: 1 is the most preferred light chain CDR1 amino acid sequence.
  • SEQ ID NO: 2 Light chain CDR 3 (NOL9-13): QVWDSSSDPVV.
  • SEQ ID NO: 2 is the most preferred light chain CDR3 amino acid sequence.
  • SEQ ID NO: 3 Light chain CDR 3 (NOL1-8): QVWDSGDGX2PRV, wherein X2 can be H or N.
  • SEQ ID NO: 4 Light chain CDR 3 (NOLI, 2, 3, 4, 6, 8): QVWDSGDGHPRV.
  • SEQ ID NO: 5 Light chain CDR 3 (NOL5 and NOL7): QVWDSGDGNPRV.
  • SEQ ID NO: 6 Heavy chain CDR 1 (NOL1-13): GFTFXsSYG, wherein Xs can be S, G orR, preferably Xs is S.
  • SEQ ID NO: 7 Heavy chain CDR 1 (NOL1-11): GFTFSSYG.
  • SEQ ID NO: 7 is the most preferred heavy chain CDR1 amino acid sequence.
  • SEQ ID NO: 9 Heavy chain CDR 1 (NOLI 3): GFTFRSYG.
  • SEQ ID NO: 10 Heavy chain CDR 2 (NOL1-13): IX4X5DGX6NK, wherein X4 can be S or W, X5 can be Y or N and X6 can be S or R, preferably X4isS,X5isYandX6isS.
  • SEQ ID NO: 11 Heavy chain CDR 2 (NOL9-12): ISYDGSNK.
  • SEQ ID NO: 11 is the most preferred heavy chain CDR2 amino acid sequence.
  • SEQ ID NO: 12 Heavy chain CDR 2 (NOL13): ISNDGSNK.
  • SEQ ID NO: 13 Heavy chain CDR 2 (NOLI, 3, 4, 5, 6, 7, 8): IWYDGSNK.
  • SEQ ID NO: 14 Heavy chain CDR 2 (NOL2): IWYDGRNK.
  • SEQ ID NO: 15 Heavy chain CDR 3 (NOL9-13): X7KGXsKIQLWFAAFDI, wherein X? can be V or A and Xs can be F or L, preferably X? is V and Xg is L.
  • SEQ ID NO: 16 Heavy chain CDR 3 (NOLI I): VKGLKIQLWFAAFDI.
  • SEQ ID NO: 16 is the most preferred heavy chain CDR3 amino acid sequence.
  • SEQ ID NO: 17 Heavy chain CDR 3 (NOL9, 12, 13): AKGFKIQLWFAAFDI.
  • SEQ ID NO: 18 Heavy chain CDR 3 (NOLI O): VKGFKIQLWFAAFDI.
  • SEQ ID NO: 20 Heavy chain CDR 3 (NOLI, NOL3-7): ARGLYGRSYFDY.
  • X ⁇ s can be D or N
  • X?9 can be Q or H
  • Xgo can be R or K
  • Xsi can be F or Y
  • X ⁇ 2 can be T
  • Aor S can be P or T
  • Xs3 can be P or T
  • Xs4 can be D or E
  • Xg5 can be F or L
  • Xg6 can be S or T
  • Xs ⁇ can be T or A
  • Xss can be S or G
  • Xs9 can be V, I, A or S
  • X9o can be G or S
  • X9i can be D or S
  • X92 can be G or D
  • X93 can be H
  • X94 can be P or V
  • X95 can be R or V
  • X96 can be V or F
  • X97 can be F or G
  • X9g can be G or T
  • X99 can be T
  • SEQ ID NO 23 Variable region light chain - full sequence (NOL9-13): QSVLTQPPSXioSVAPGQTARITCGGNNIGSKSVHWYXiiQKPGQAPVL VVYDDSDRPSGIPERFSGSNXi2GNTATLTIXi3RVEXi4GDEADYYCQV WDSSSDPVVFGGGTKLTVL, wherein Xio can be V or I, Xii can be Q or H, Xi2 can be S or T, Xis can be S or G and Xi4 can be A or S, preferably Xio is V, Xi 1 is Q, Xi2 is S, Xi3 is S and Xi4 is A.
  • SEQ ID NO: 24 Variable region light chain - full sequence (NOLI 1, NOL9): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL SEQ ID NO: 24 is the most preferred variable region light chain full sequence.
  • SEQ ID NO: 25 Variable region light chain - full sequence (NOL 1 -8): SYELTQPPSXi5SVAPGQTAXi6lXi7CEGDNIGSKSVHWYQQRPGQAPV LVVXi8DDXi9DRPSGIX2oDRX2iSGSNSGNTAX22LTISRVEX23GDEADY YCQVWDSGDGX24PRVFGGGX25KLTVL, wherein Xi5 can be M or V, Xi6 can be K or R, Xi7 can be A, V or T, Xi8 can be F or Y, Xi9 can be T or A, X20 can be T or P, X2i can be L or F, X22 can be T or A, X23 can be V or I, X24 can be H or N and X25 can be T or S.
  • SEQ ID NO: 26 Variable region light chain - full sequence (NOLI): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL.
  • NOLI Variable region light chain - full sequence
  • NOL2 Variable region light chain - full sequence
  • SEQ ID NO: 28 Variable region light chain - full sequence (NOL3): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRLSGSNSGNTAALTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL.
  • SEQ ID NO: 29 Variable region light chain - full sequence (NOL4): SYELTQPPSVSVAPGQTAKIVCEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL.
  • SEQ ID NO: 30 Variable region light chain - full sequence (NOL5): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDADRPSGITDRFSGSNSGNTATLTISRVEIGDEADYYCQVWDSGD GNPRVFGGGTKLTVL.
  • SEQ ID NO: 31 Variable region light chain - full sequence (NOL6): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGSKLTVL.
  • SEQ ID NO: 32 Variable region light chain - full sequence (NOL7): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GNPRVFGGGTKLTVL.
  • SEQ ID NO: 33 Variable region light chain - full sequence (NOL8): SYELTQPPSVSVAPGQTAKITCEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL.
  • SEQ ID NO: 35 Variable region light chain - full sequence (NOL10): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTIGRVESGDEADYYCQVWDSSSD PVVFGGGTKLTVL.
  • SEQ ID NO: 36 Variable region light chain - full sequence (NOL12): QSVLTQPPSISVAPGQTARITCGGNNIGSKSVHWYHQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL.
  • SEQ ID NO: 37 Variable region light chain - full sequence (NOL13): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNTGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL.
  • Xss can be D or A
  • X59 can be Y or A
  • X6o can be W or F
  • X6i can be G or D
  • X62 can be Q or I
  • X63 can be G or W
  • X64 can be T or G
  • X65 can be L or Q
  • X66 can be V or G
  • X6? can be T or A
  • X ⁇ s can be V or M
  • X69 can be S or V and X?o can be S or T.
  • SEQ ID NO: 39 Variable region heavy chain - full sequence (NOL9-13): EVQLX26ETGGGVVX27PGRSLRLSCAASGFTFX2gSYGMHWVRQAPGK GLEWVAVISX29DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCX3oKGX3iKIQLWFAAFDIWGQGX32MVTVSS, wherein Xi6 can be E or V, X2?
  • X28 can be S, G or R, Xi9 can be Y or N, Xso can be V or A, X3i can be F or L and Xs2 can be T or A, preferably Xi6 is V, X27 is Q, X28 is S, X29 is Y, Xso is V, X3i is L and Xs2 is T.
  • SEQ ID NO: 40 Variable region heavy chain - full sequence (NOLI I): EVQLVETGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCVKGLKIQLWFAAFDIWGQGTMVTVSS.
  • SEQ ID NO: 40 is the most preferred variable region heavy chain full sequence.
  • Variable region heavy chain - full sequence (NOL 1 -8): QVQLQQSXssGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGL EWX34AVIWYDGX35NKYYADSVKGRFTISRDNSKNTLYLQX36NSLRA EDTAVYYCARGLX37GRSYFDYWGQGTLVTVSS, wherein Xss can be E or G, Xs4 can be L or V, Xs5 can be S or R, Xs6 can be M or I and Xs? can be Y or F.
  • SEQ ID NO: 42 Variable region heavy chain - full sequence (NOLI, NOL3, NOL5, NOL6, NOL7): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLYGRSYFDYWGQGTLVTVSS.
  • SEQ ID NO: 43 Variable region heavy chain - full sequence (NOL2): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGRNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLFGRSYFDYWGQGTLVTVSS.
  • SEQ ID NO: 45 Variable region heavy chain - full sequence (NOL4): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQINSLRAEDTAV YYCARGLYGRSYFDYWGQGTLVTVSS.
  • Variable region heavy chain - full sequence (NOL8): QVQLQQSEGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WLAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLFGRSYFDYWGQGTLVTVSS.
  • SEQ ID NO: 50 Variable region heavy chain - full sequence (NOL9): WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGTMVTVSS.
  • Variable region heavy chain - full sequence (NOL 10): EVQLVETGGGVVRPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE VYYCVKGFKIQLWFAAFDIWGQGTMVTVSS.
  • SEQ ID NO: 52 Variable region heavy chain - full sequence (NOL 12): EVQLEETGGGVVQPGRSLRLSCAASGFTFGSYGMHWVRQAPGKGLE WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGTMVTVSS.
  • SEQ ID NO: 53 Variable region heavy chain - full sequence (NOL13): EVQLVETGGGVVQPGRSLRLSCAASGFTFRSYGMHWVRQAPGKGLE WVAVISNDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGAMVTVSS.
  • SEQ ID NO: 54 Light chain - full sequence (NOLI I): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVLGPTSTPSVSLFPPSSEELSANKATVVCLISDFSPSGL EVIWKVNDAVTTDGVQTTRSSKQSNGKYAASSYLTRTSAQWKSYSS VSCQVKHQGKTVEKKLSPSECP.
  • SEQ ID NO: 54 is the most preferred light chain full sequence.
  • SEQ ID NO: 55 is the most preferred heavy chain full sequence.
  • SEQ ID NO: 56 Constant region light chain - full sequence: GPTSTPSVSLFPPSSEELSANKATVVCLISDFSPSGLEVIWKVNDAVTT DGVQTTRSSKQSNGKYAASSYLTRTSAQWKSYSSVSCQVKHQGKTV EKKLSPSECP.
  • Nucleic acid sequence heavy chain CDR1 NOL2 GGATTCACTTTCAGTAGCTATGGC SEQ ID NO: 69
  • Nucleic acid sequence heavy chain CDR1 NOL12 GGATTCACCTTCGGTAGCTATGGC SEQ ID NO: 70
  • Nucleic acid sequence heavy chain CDR2 NOL2 ATATGGTATGATGGAAGTAATAAA SEQ ID NO: 79
  • SEQ ID NO: 156 Light chain CDR 2 (NOLI 1,NOL9,NOL10,NOL12,NOL13): DDS. ID NO: 156 is the most preferred light chain CDR2. SEQ ID NO: 157 Light chain CDR 2 (NOLI, 2, 3, 4, 6, 7, 8): DDT. SEQ ID NO: 158 Light chain CDR 2 (NOL5): DDA.
  • SEQ ID NO: 159 Nucleic acid sequence variable region light chain CDR2 (NOLI, NOL2, NOL3, NOL4, NOL6, NOL7, NOL8): GATGACACC SEQ ID NO: 163
  • Nucleic acid sequence variable region light chain CDR2 NOL5 GATGACGCC
  • Nucleic acid sequence variable region light chain CDR2 (NOLI 1, NOL9, NOL10, NOL12, NOL13): GATGATAGC.
  • the isolated antibody or antibody derivative may comprise at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 1, 155, 156, 157, 158,2,3,4,5,6,7,8,9,10, 11,12,13,14,15, 16, 17, 18, 19, 20 and 21.
  • the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155,156,2,6,7, 10, 11, 15 and 16.
  • the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155, 2, 6, 10 and 15.
  • the isolated antibody or antibody derivative comprises all complementary determining region (CDR) amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155,2,6,10 and 15.
  • the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 156, 2, 7, 11 and 16.
  • the isolated antibody or antibody derivative comprises all complementary determining region (CDR) amino acid sequences as depicted in any one ofSEQ ID NOs: 1, 156, 2, 7, 11 and 16.
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155, 156, 157 and 158, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3, 4 and 5, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6, 7, 8 and 9, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs:
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155, 156 and 157, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3 and 4, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6 and 7, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 10, 11 and 13, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID N0s:15, 16, 17, 19 and 20.
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 156 and 157, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2 and 4, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in SEQ ID NO: 7, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 11 and 13, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 16, 17 and 20.
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence as depicted in SEQ ID NO: 155, a light chain CDR3 amino acid sequence as depicted in SEQ ID NO: 2, a heavy chain CDR1 amino acid sequence as depicted in SEQ ID NO: 6, a heavy chain CDK2 amino acid sequence as depicted in SEQ ID NO: 10 and a heavy chain CDR3 amino acid sequence as depicted in SEQ ID N0:15.
  • the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence as depicted in SEQ ID NO: 156, a light chain CDR3 amino acid sequence as depicted in SEQ ID NO: 2, a heavy chain CDR1 amino acid sequence as depicted in SEQ ID NO: 6, a heavy chain CDR2 amino acid sequence as depicted in SEQ ID NO: 11 and a heavy chain CDR3 amino acid sequence as depicted in SEQ ID N0:16.
  • the isolated antibody or antibody derivative optionally comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23, 24, 25, 26,27,28,29, 30, 31, 32,33,35,36 or 37, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39, 40, 41, 42,43, 45, 49, 50, 51, 52 or 53.
  • the isolated antibody or antibody derivative comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23 or 24, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39 or 40.
  • the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23 or 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39 or 40.
  • the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in SEQ ID NO: 23 or 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in SEQ ID NO: 39 or 40.
  • the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in SEQ ID NO: 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in SEQ ID NO: 40.
  • the isolated antibody or antibody derivative according to the invention comprises a light chain amino acid sequence as depicted in SEQ ID NO: 54, and/or comprises a heavy chain amino acid sequence as depicted in SEQ ID NO: 55 or 153, more preferably as in SEQ ID NO: 55.
  • the invention also provides isolated nucleic acid molecules, vectors and host cells comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to the invention.
  • the at least one nucleic acid sequence is selected from the group consisting of the nucleic acid sequences as depicted in any one ofSEQ ID NO: 58, 59,69,70,71,72, 79,83, 84, 85, 91, 92, 93, 94, 97, 103, 105, 109,110,111, 114, 115, 116, 118,123, 124,125,126, 128, 130-150, 152, 154, 159, 163 and 167.
  • the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO: 58, 79,94,105,118,133,146, 149,150,152,154 and 167. More preferably, the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO: 58, 79, 94, 105,118 and 167.
  • the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO:133,146,149,150 and 152.
  • all ofSEQ ID NO: 133, 146, 149 and 150 are used to produce the antibody or antibody derivative of the present invention on one or more vectors or in on or more host cells.
  • the nucleic acid sequences SEQ ID NOs: 149 or 152 and 150 encode the constant regions of the heavy and light chain of the antibody or antibody derivative according to the invention, respectively.
  • truncated constant regions and, thus, truncated forms of SEQ ID NOs: 149, 150 and/or 152 may be used.
  • NOLI 1 is the most preferred antibody.
  • a method of manufacturing of the isolated antibody or antibody derivative according to the invention comprising expressing the isolated antibody or antibody derivative in a host cell.
  • the isolated antibody or antibody derivative is expressed in a host cell according to the invention, such as a eukaryotic cell.
  • the isolated nucleic acid molecules, vectors and host cells comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to the invention as described above are particularly useful in the method of manufacturing of the isolated antibody or antibody derivative according to the invention and may be used for expressing the antibody or antibody derivatives.
  • BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Titration ELISA of affinity-maturated antibodies from wildtype NOLI on eqIL-5.
  • Figure 2 Titration ELISA of affimty-maturated antibodies from wildtype NOL2 oneqIL-5.
  • Figure 3 Inhibition effect of antibodies on the binding of eqIL-5 to its receptor.
  • Figure 6 Reducing (left) and non-reducing (right) SDS-PAGE of samples of NOLI 1 after storage at 4 °C, 21 °C and 37 °C.
  • Figure 7 SDS-Page stained with Coomassie (left) and Western Blot stained with Streptavidin-HRP (right) of NOLI 1, produced in Hi5 insect cells and Expi293FTM mammalian cells. Aglycosylated samples containing the N297A mutation and glycosylated samples were analyzed.
  • Figure 8 Timeline of the target animal study D10POC-A. Three horses received the antibody (42.62
  • the average AOD450nm-620nm signal of the three horses of the antibody group was compared to the average signal of the three horses of the control group at a serum dilution of 1:100.
  • Figure 10 Timeline of the target animal studies D10POC-B and D10POC-C. Injection of NOLI 1 took place at day 0, 30 and 60.
  • Figure 11 Anti-Dmg antibody (ADA) assay of target animal study D10POC-B: Evaluation ofAnti-Drug-Antibody formation in the horses treated with NOL11.
  • the two-center study D10POC-B took place in Eastern and Northern Germany. The study in Eastern Germany is still on going, therefore, only the data from the Northern Germany study is included.
  • the antibody group consists of two horses treated with NOL11.
  • the control group consists of two horses that were not treated with the antibody. Injection of NOLI 1 took place at day 0, 30 and 60.
  • Figure 12 Skin scores of insect bite hypersensitivity (IBH) affected horses (study D10POC-C, day -7-91). The severity ofIBH was determined for each horse before the trial (mild, moderate, or high grade IBH). Data show the skin scores between 0 (no abnormality) and 3 (severe itch/skin lesions) evaluated during the 91-day study period (starting 7 days prior the first injection). The horses were scored in the first five days after each injection and then every second day.
  • Figure 14 Skin scores of insect bite hypersensitivity (IBH) affected horses (study D10POC-C, day 91 -135). The severity ofIBH was determined for each horse before the trial (low, moderate, or high grade IBH). Data show the skin scores between 0 (no abnormality) and 3 (severe itch/skin lesions) evaluated during day 91 and day 135 after the last injection of NOL11. The horses were scored every second day. The antibody (0.05 mg/kg) was administered subcutaneously at three time points (day 0, day 30 and day 60). (a) Skin scores of the untreated control horse (No.329) classified as low to moderately affected by IBH. (b) Skin score of the moderately affected horse No.344 treated with NOLI 1.
  • IBH insect bite hypersensitivity
  • FIG. 15 The graph shows the linear regression analysis of the skin score of the three study horses evaluated between day 91 and day 135 after the last injection of NOLI 1.
  • the antibody 0.05 mg/kg was administered subcutaneously at three time points (day 0, day 30 and day 60).
  • the severity of IBH was determined for each horse before the trial (mild, moderate, or high grade IBH).
  • the equine pruritus visual analogue scale (EPVAS) was used to determine the scratching behavior.
  • the scale is graduated from 0 (normal horse; no itch) to 10 (extremely severe itching).
  • the antibody (0.05 mg/kg) was administered subcutaneously at three time points (April, May and June; four weeks between each in injection). The observation started 21 days after the last injection of NOLI 1 (theoretical half-life of equine antibodies).
  • Figure 17 Overview CDR1, CDR2 and CDR3 of the variable light chains of NOL1-13.
  • Figure 18 Overview CDR1, CDR2 and CDR3 of the variable heavy chains of NOL1-13.
  • Monoclonal antibodies against the summer eczema of horses Monoclonal antibodies against eqIL-5 were generated via antibody phage display. Afterwards, the format of the binders was changed from single chain variable fragment (scFv) to full length immunoglobulin G (IgG). For the constant domains, an equine sequence was used. Out of the seven equine IgG subclasses, the subtype IgG6 was chosen due to its lowest effector function.
  • the antibodies were produced in mammalian cells (Expi293FTM) and validated by Enzyme-linked Immunosorbent Assay (ELISA) and a self-established cellular inhibition assay using flow cytometry.
  • Heavy and light chain of the antibody were cloned into different expression plasmids which were co-transfected in a ratio of 1:1.
  • the production scale was adjusted depending on the required protein amount and ranged between 7.5 mL to 125 mL. Independent of the scale, between 1.5 - 2.0 x 106 cells/mL were used. They were transfected with plasmid DNA and PEI per milliliter of cell culture volume. DNA and PEI were first diluted separately and then mixed in F 17 medium (10% of the culture volume) and incubated at room temperature for around 30 min. Then, the DNA-PEI mix was added to the cells. The cells were cultivated under the conditions mentioned above for 7 days in total, including a feeding step after 48 h with new medium.
  • the cells were harvested via centrifugation. Subsequently, the supernatant was sterile filtered for purification. Antibodies were purified from the culture supernatant using affinity chromatography via binding to protein A or protein G. Depending on the scale, different columns were installed using the Akta or Profinia system or a 24-well filter plate and compressed air was utilized. 7.7.2 Titration ELISA The binding of the antibodies to eqIL-5 was detected by ELISA. For this purpose, eqIL-5 was immobilized in a 96-well plate, the corresponding antibodies were added and then detected with an antibody directed against the equine Fc part. As a negative control for non-specific binding, the antibody was also added to BSA.
  • the reaction was stopped with sulfuric acid and the optical density was measured at 450 mn and 620 nm with an ELISA reader.
  • a titration from 316 nM to 0.0036 nM with the dilution factor 1/ ⁇ /10 was performed and then the optical density (OD450nm- OD620nm) was plotted against the antibody concentration.
  • the measurements were performed in triplicates.
  • a fit with the "logistic5" function using the software OriginPro was used to determine EC50 values. 1.1.3
  • Cellular inhibition assay A cellular iiAibition assay was developed in order to test whether the selected antibodies not only bind the antigen but can also inhibit the binding of the interleukin to its receptor.
  • Equine eosinophil cells would have been suitable for this purpose, as they express the receptor for eqIL-5.
  • Expi293FTM suspension cells were used instead and transiently transfected with the receptor DNA.
  • the receptor was not secreted due to a fusion to the transmembrane domain, but presented on the cell surface.
  • the eqIL-5 receptor consists of two subunits, the IL5RA and CSF2RB, which were co-transfected.
  • the cells were co-transfected with eGFP to differentiate transfected from non-transfected cells. Transfection was performed as described for antibody production. Cells were then cultured for 48 h at 37 °C, 110 rpm and 5% C02. After 48 h, the cells were used for the inhibition assay. The measurements were performed in the flow cytometer.
  • the antigen was fluorescently labelled via an anti His-tag antibody and a secondary detection antibody that is fluorescently labelled. Thus, binding of the antigen could be visualized by a fluorescent cell. A decrease in fluorescence indicated inhibition of binding.
  • a background measurement was performed using neither antigen nor inhibiting antibody, as well as a reference measurement with antigen but no inhibiting antibody.
  • the antibodies to be tested were diluted to a concentration of 2
  • the antigen was added to the antibodies in the same volume and at a concentration of 20 nM, so that the antibody had a final maximum concentration of 1 pM and the antigen a final concentration of 10 nM.
  • the antibody and antigen were preincubated for 45 minutes at room temperature.
  • cells transfected with receptor DNA and eGFP were pelleted and adjusted to a cell density of5xl06 cells/mL in FACS buffer.
  • 5 x 105 cells/well were placed in a 96-deepwell plate and washed with FACS buffer.
  • the cell pellet was resuspended in the antibody-antigen mix and incubated on ice for about 1 hour.
  • the cell pellet was taken up in primary antibody (Penta His antibody: Qiagen, 3460, 1:50 dilution) and again incubated on ice for about 1 h.
  • the reference indicates the APC value, which is generated by the binding of the antigen (eqIL-5-His) to the cells without inhibition. In the evaluation, this value corresponds to 100% binding. In the case of an inhibitory antibody, this prevents the antigen from binding to the cell, which reduces the APC signal accordingly.
  • These APC values are related to the 100% of the APC value of the reference control and thus give the relative binding of the antigen to the cell in percent. The more inhibitory the antibody, the lower the relative binding. A titration of the antibodies was performed to determine the relative binding as a function of antibody concentration. The IC50 value, which characterizes the antibody concentration at 50% inhibition, provides a suitable benchmark.
  • Table 1 EC50 values of the antibodies against eqIL-5 determined by titration ELISA The values are all in a similar, nanomolar range. The lowest value is 0.44 nM, the highest 8.99 nM. For the NOLI antibody the affinity maturation improved the £ €50 value by factor 2.2 and for the NOL9 antibody the EC50 value was improved by factor 1.4. 7.2.2 Cellular inhibition assay Since the titration ELISA only informs about the binding of the antibodies to the antigen, an additional assay to test the inhibition was performed. The goal was to analyze the antibodies' ability to inhibit the binding of the antigen to its receptor on a cell surface.
  • Figure 3 presents the inhibition effect of the antibodies against eqIL-5.
  • the y-axis describes the relative binding of the interleukin to its receptor.
  • the antibodies partially inhibit the binding completely, depending on their concentration.
  • the curves of the different antibodies are very similar.
  • the antibodies derived after affinity maturation of both wildtypes are equally effective in inhibition, despite slightly different affinities in the titration ELISA ( Figures 1 and 2). This is possibly explained by the fact that the antibodies recognize different epitopes.
  • the curves were determined with the "logistic 5" fit using the OriginPro software.
  • Table 2 shows the IC5o values which are the antibody concentrations needed to reduce the binding to 50%. A reduction to 50% binding is achieved at concentrations between 3.0 nM and 9.4 nM.
  • the affinity maturation improved the IC50 value by factor 2.0 and for the NOL9 antibody the IC50 value was improved by factor 1.9.
  • Table 2 IC50 values of the antibodies against eqIL-5 determined by cellular inhibition assay 2. Characterization of a chimeric-equine anti-ILS antibody Further experiments were performed with NOLI 1, a preferred antibody. 2.1 Materials and Methods 2.1.1 Stability tests To determine the long-term stability of the antibody NOLI 1, different aliquots were stored at 4°C, 21°C and 37°C. After two weeks of storage, new aliquots were added. After two additional weeks, the samples were analyzed by HPLC-SEC, ELISA and SDS-PAGE in comparison with freshly thawed material to obtain data for two and four weeks of storage at the given temperatures.
  • HPLC-SEC the antibodies were diluted to a concentration of 50
  • N297A The aglycosylation mutation is called N297A based on the fact that human antibodies are glycosylated at the position 297.
  • a biotinylation experiment was performed.
  • a glycosylated version of NOLI 1 was used as a positive control. This experiment was performed with NOLI 1 recombinantly produced in Expi293FTM cells as described previously and Hi5 cells (described below).
  • the antibodies were prepared by changing the buffer via gel filtration to 100 mM NaAc/150 mM Nad, pH 5.5. Then they were incubated in sodium-periodate solution for oxidation of sialic acid, a common sugar component of protein polysaccharides. The buffer was again changed by gel filtration. As the next step Hydrize-PEG4-Biotin was added to the samples (final cone.5 mM) and incubated. Excess biotin was removed by gel filtration and the samples were used for Western Blot Analysis. 2.7.2.7 Western Blot Analysis In Western Blot analysis, the biotin was specifically detected. After a reducing SDS- PAGE was performed, the proteins were blotted onto a membrane.
  • DNA and PEI were directly added to the cells.
  • the cells were diluted to a concentration of 1.0 x 106 cells/mL and cultivated under the conditions mentioned above for 4 days in total, including a feeding step after 48 h with new medium.
  • the cells were harvested via centrifugation. Subsequently, the supernatant was sterile filtered for purification.
  • Antibodies were purified from the culture supernatant using affinity chromatography via binding to protein A or protein G. Depending on the scale, different columns were installed using the Akta or Profinia system or a 24-well filter plate and compressed air was utilized.
  • the aggregates are also present in the fresh sample.
  • the antibody When the antibody is stored at 37 °C, slightly more aggregated antibody can be detected. Under reducing conditions in all analyzed samples, two bands are visible at 25 kDa and 50 kDa. The disulfide bonds between the heavy and light chain of the antibody are broken and we see one band for the heavy chain (50 kDa) and one band for the light chain (25 kDa).
  • the sample, which was stored at 37 °C shows two slight additional bands and around 100 kDa and 150 kDa. No big differences are visible between the samples, leading to the conclusion, that the antibody is stable under all tested conditions (Figure 6).
  • NOLI 1 Due to promising in vitro results in the inhibition assay as well as producibility and stability data, the anti-eqIL-5 antibody NOL11 tested in vivo in horses (healthy and IBH affected). NOLI 1 has been tested for safety and tolerability in two target-animal studies to date (D10POC-A and D10POC-B). In the second study, the efficacy of the antibody was additionally tested in a small number of animals suffering from IBH (D10POC-C). An overview of each study with relevant information on the animals involved is provided in Table 3. Two healthy control animals that received placebo or were untreated are not included in Table 3.
  • Table 3 Designation of the target-animal studies A, B and C with NOL11 BW: Body weight; IBH: insect bite hypersensitivity.
  • D10POC-A single-center safety study (Northern Germany) / orientation study A
  • D 1 OPOC-A six horses from the own herd at the WDT serum factory in Memsen, Germany, were used. The primary aim of the study was the safety evaluation of the antibody NOL11. Therefore, four healthy horses and two horses affected by IBH were used. The horses were divided in an antibody and a control group each containing two healthy and one sick horse. The control group received 1 x PBS instead of the antibody.
  • An overview of the test horses is given in Table 3.
  • horse No.306 had a total score of 3 and horse No.329 a total score of 1. This time point is outside the time period, in which the antibody influences the symptoms.
  • horse No.306 is stronger affected by summer eczema as it was the case in the previous years.
  • Table 5 shows a comparison of the summer eczema scores and symptoms of former years with the symptoms of the study year (after treatment).
  • Table 5 Comparison of severity of summer eczema symptoms of horse No.306 and No.312 Taken together, there was no observation of adverse side effects of the NOL11 antibody. Therefore, a high safety and tolerability is assumed for this product. Concerning the efficacy, one has to mention that in the study year in general the symptom development of horses with summer eczema seemed to be reduced. Furthermore, the number of the used horses with summer eczema in this study does not allow statistically significant evaluation of the efficacy. Nevertheless, taken the fact that horse No.306 had more severe symptoms over the last years compared to horse No.329, one can state a tendency of efficacy of NOLI 1 treatment since the symptoms of the two horses were similarly low during the study period.
  • ADA Anti-drug-antibody
  • Binding of serum antibodies to the Fc-part of the antibody is not expected since the Fc-part is a constant region that is part of the equine immune system) were coated per well in a high-binding 96-well plate at 4 °C overnight.
  • an alternative eqFab antibody and 2% BSA were used.
  • wells were blocked with 2% BSA in PBS-T for 1 h at room temperature and washed three times using an ELISA washer (BioTekTM 405TM LS).
  • the horse serum was titrated with a starting dilution of 1/10 and a dilution factor of 1/VTH and incubated for 1 h at room temperature.
  • the detection antibody goat IgG anti horse IgG (Fc-)HRP (SAB3700145, Lot RI36383, Sigma- Aldrich, dilution 1:20000 in 2% BSA) was applied for 1 h at room temperature. Wells were washed again and TMB substrate was added to start the coulometric reaction and 1 M sulfuric acid was used to stop the reaction. The absorbance (OD450nm-620nm) was measured with an ELISA reader (Epoch, Biotek). All measurements were performed in duplicates. For the analysis, the signals of the serum sample day -1 were subtracted from the signal of all other serum samples (AOD450nm-620nm) so that only the change but not the absolute value is evaluated.
  • the average AOD450nm-620nm signals were compared at a serum dilution of 1:100 ( Figure 9).
  • the AOD450nm-620nm signal from day 14 onwards is increased up to approximately 0.06 while the signal of the control group fluctuates closely around 0. This is still in the expected fluctuation range and does not indicate the development of ADAs. If the horse generated ADAs it would happen after the second dose with some delay in time. Up to 120 days no significant increase of the signal of the antibody group compared to the control group was detected. The slight signal of the antibody group from day 14 on is still in a range that lies within the expected fluctuations.
  • Horse No.306 was a part of the studies D10POC-A and D10POC-B and C, therefore receiving the antibody a second year in row. As the likelihood of ADA formation may increase with the number of injections, horse No.306 was carefully screened for adverse immune reactions. The study in Eastern Germany is still on-going, therefore, only the samples from the horses in Northern Germany were analyzed so far.
  • the AOD450nm-620nm signal of day 90 and day 91 is increased up to approximately 0.09 and 0.10 while the signal of the control group fluctuates closely around 0. This is still within the expected fluctuation range and does not indicate the development ofADAs.
  • the signal of the antibody group decreases in the same range to approximately -0.09, showing that this is a normal fluctuation range.
  • Horse No.306 showed no immune response to of NOLI 1 the second year in a row with a total of five injections (two in D10POC-A, three in D10POC-B). This indicates that the antibody is safe for a repeated treatment of horses.
  • study D10POC-C Single-center efficacy study (Northern Germany) 3.3.1 Study design During the study D10POC-B, two horses treated with NOLI 1 as well as an untreated control horse were intensively monitored for the expression of symptoms (referred to as study D10POC-C). The assessment was carried out by the veterinary who has cared for the horses for several years and knows their medical history very well. Before the study started, the severity of IBH of the study horses was assessed, based on clinical expression in previous years.
  • Horse No.344 (Fig.12b) had an average score of 0.1 between day 0 and day 91, whereas the average score of the control horse (horse No.329, Fig.12a) was 1.0.
  • the average score for severely affected horse No.306 was 0.7 and therefore below the score of the control horse.
  • Fig.13 shows the regression line of the scores over the 91-day observation period. The increase in the untreated horse is the most prominent, while the line of the moderately affected horse hardly increases. The increase in the severely affected horse treated with NOLI 1 is lower than for the untreated control animal.
  • Fig.16 shows the development of scratching behavior 21 days after the last injection of NOLI 1 (dose 3). Scratching behavior was scored from 0 to 10 using the EPVAS score. In the untreated control horse as well as in the NOL 11-treated highly affected IBH horse, pruritus increases constantly 21 days after the last dose. However, the NOL 11-treated horse with severe IBH (No.306) did not show more severe pruritus than the untreated horse, known to have mild to moderate disease, even 81 days after the last antibody injection. Horse No.344 started to scratch 39 days after the last NOLI 1 injection but showed only minor scratching behavior until the end of the study.
  • the horse was not treated with repellents, skin oil or glucocorticoids.
  • study D10POC-A the horse received two injections of NOL11 (0.04 mg/kg bodyweight) two weeks apart (April and May) before the onset of first symptoms.
  • NOL11 0.4 mg/kg bodyweight
  • the horse showed slight chafing around the mane crest and sheared hair in the area of the tail rump.
  • approximately 80 days after the last NOL11 injection the expression of the symptoms became somewhat more marked, and the horse showed more distinct rubs in the mane and tail area as well as some hairless areas on the belly suture. However, there were still no visible skin lesions or wounds.
  • the horse did not require any further protective or therapeutic measures during the test season (no repellents, stabling, reduced grazing, regular stabling, skin oil or glucocorticoids) and still showed a significantly lower expression of summer eczema compared to previous years.
  • the horse received three injections of NOLI 1 (0.05 mg/kg bodyweight; 30 days interval in April, May and June) after the onset of symptoms. This was done to see if it is possible to down-regulate an already existing symptomatic condition. The horse showed only minor symptoms until mid-June. At this time point, the horse showed more pronounced rubbing and was treated with skin oil for two consecutive days. After the third NOL11 injection, the symptoms weakened again.
  • the horse is moderately affected by IBH without protective measures (stabling for most of the day, reduced grazing, daily application of skin care oil and regular application of the repellent permethrin).
  • the horse shows increasing chafing over the season, especially on the mane crest and tail rump.
  • the treatment consists of weekly application of permethrin (WellcareTM Emulsion 10.5 mg/ml) and care of the affected areas with skin oil. No glucocorticoid treatment is needed.
  • Horse 2 participated only in studies D10POC-B and C. Analogous to horse 1 (chapter 3.4.1), horse 2 received three injections ofNOLll (0.05 mg/kg bodyweight; 30 days interval in April, May and June) after the onset of symptoms.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biophysics (AREA)
  • Dermatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present invention provides an isolated antibody or antibody derivative that specifically binds to an equine interleukine-5 (eqIL-5) comprising at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof. Further provided are an isolated nucleic acid molecule comprising a nucleic acid sequence encoding the isolated antibody or antibody derivative, a vector comprising the respective nucleic acid sequence and a host cell comprising the isolated nucleic acid molecule. The present invention also provides a pharmaceutical composition comprising the isolated antibody or antibody and a pharmaceutically acceptable excipient. In addition, the use of the isolated antibody or antibody derivative or pharmaceutical composition as a medicament is provided, in particular, the use in the treatment of an allergic condition of a horse. The present invention also relates to in vitro methods for the diagnosis of an allergic condition in a horse as well as to methods of manufacturing the isolated antibody or antibody derivative.

Description

Wirtschaftsgenossenschaft deutscher Tierarzte eG Munchen, 17 October 2023 Our reference: W2260WO Wirtschaftsgenossenschaft deutscher Tierarzte eG SiemensstraBe 14, 30827 Garbsen, Germany — Antibodies and antibody derivatives against equine IL-5 DESCRIPTION Allergic conditions affect about 10% of horses (including horses, ponies, donkeys, hinnies and mules) worldwide. Among those, seasonal allergies are the most common and they mainly belong to the group of skin allergies and respiratory allergies. The most prominent examples of skin and respiratory allergies are insect-bite hypersensitivity and asthma. Insect-bite hypersensitivity (IBH, also denoted as Culicoides hypersensitivity (CH), sweet itch, summer eczema, kasen, Queensland itch, summer dermatitis or equine dermatitis) is an allergic reaction to bites from mosquitos and midges (Culicoides), mainly percutaneously. This most common allergic dermatitis may affect up to 72% of allergic horses (including ponies) depending on race and region, especially in the warmer regions, and is a reaction of the animal to the saliva of midges (Culicoides) and other insects. The bites lead to an allergic reaction with a massive infiltration of eosinophil cells to the allergic skin area. The horses suffer of pruritus, skin lesions, hair loss and secondary infections, which can lead to chronic dermatitis. They experience itchiness, which is associated with the insect bite (pruritus), and this can lead to cases, where the animals scratch that strong so they hurt themselves leading to open skin lesions with alopecia and crusting. These lesions may then promote secondary infections. Thus, horses severely affected from summer eczema cannot live a normal herd life. Horses with asthma suffer from chronical airway inflammation that causes a poor performance to a cough, nasal discharge or overt respiratory distress. Asthma ("equine asthma syndrome") includes conditions ranging from mild asthma, also called inflammatory airway disease (IAD) in young horses, to severe asthma, widely referred to as heaves (chronic obstructive pulmonary disease (COPD) or recurrent airway obstruction (RAO)) and pasture- or summer-associated airway disease. The clinical signs primarily develop after exposure to aerosolized particles, which could be endotoxins produced by specific bacteria, particles originating from molds, fungi, beta-D-glucan, microorganisms, vegetative material, inorganic dusts, and noxious gases, such as ammonia from urine, mite debris and other. Severely asthmatic horses are usually managed through antigen avoidance and the use of corticosteroids and bronchodilators to reduce airway inflammation, bronchoconstriction and improve lung function (Simoes, J. Equine Vet. Sci.2020, 87, 102937). However, some horses are unresponsive to corticosteroid treatment and an effective treatment is still needed. Like with other allergic conditions, also in asthma eosinophilic airway inflammation is observed. Interleukin-5 (IL-5) is connected to various chronic inflammatory conditions and seems to be involved in allergic conditions, including equine allergic asthma and insect-bite hypersensitivity, which is the most common allergic condition in horses affecting the skin (Janssen et al.. Frontiers in Immunology 2022, 13, 921077, Roufosse, Front. Med.,5,49, Fettelschoss-Gabriel etal.. The Veterinary Journal 1Q11, 276, 105741). As reported in Fettelschoss-Gabriel et al. (2021), equine IBH shows a type I allergy phenotype, including allergen-derived CD4 type-2 T helper (Th2) polarization and IL-4/IL-13 predominance. This is explained to lead to Culicoides allergen-specific IgE and corresponding degranulation of mast cells and basophiles. Eosinophilia has been suggested to play an increasingly dominant role in chronic allergic disease phases, whereas early phases are dominated by Type I-IgE mediated reactions (Fettelschoss- Gabriel et al.. The Veterinary Journal 2021,276, 105741). Upon chronic exposure to allergens, a shift from conventional Th2 (cTh2) cells towards pathogenic effector Th2 (peTh2) cells occurs. The latter secrete high levels of IL-5, leading to eosinophilia (Mitson-Salazar and Prussin, Frontiers in Medicine 2017, 4, 165). IL-5 is the most potent activator ofeosinophils and is produced by Th2 cells and ILC2s in mammals. IL-5 is also reported to have a role in eosinophil extracellular trap cell death (Nagase et al., Allergology International 2020, 69, 178-186). It binds to the interleukin-5 receptor and is, therefore, involved in B-cell growth and immunoglobulin secretion. As a key mediator in eosinophil activation, IL-5 was identified by the inventors as a target for addressing allergic conditions, such as IBH, atopic dermatitis and asthma, in horses. The important role of eosinophils in the molecular mechanism ofatopic dermatitis in horses can be seen e.g., in Fettelschoss-Gabriel et al. (2021). IL-5 was also shown to be upregulated in other allergic horses and, in particular, in horses with asthma (Janssen et al.. Frontiers in Immunology 2022, 12, 921077). Dewachi et al. (2006) found out that horses with heaves had significantly increased numbers ofneutrophils expressing IL-5 receptors compared to control while in pasture, and further increased during stabling in heaves affected horses but not in control animals (Dewachi et al., Vet Immunol Immunopathol 2006, 109(1-2), 31-6). Using immunohistochemistry and in situ hybridization, the expression ofIL-5 was observed in the BALF lymphocytes of sEA-affected horses (Cordeau et al., Vet. Immunol. Immunopathol.2004, 97, 87-96, Lavoie et al., Am. J. Respir. Crit. Care Med.2001, 164, 1410-1413). Bond et al. (2019) demonstrated a down-regulation of IL-5 in response to dexamethasone administration in horses with mild asthma (Bond et al., BMC Vet Res. 2019, 15, 397). Others reported that IL-5 is upregulated in horses with mild equine asthma, in both mastocytic and neutrophilic phenotypes (Beekman et al. , J Vet Intern Med.2012,2^(1), 153-161. This seems to be different to the findings in humans, where IL-5 is highly specific for eosinophilic inflammation. Whilst eosinophils are less commonly detected in equine BALF, excepting a sub-group of mild equine asthma (MEA) reported predominantly in young horses associated with dust exposure (Riihimaki et al.. Can J Vet Res.2008, 72(5), 432, Ivester et al., J Vet Intern Med. 2014, 28(3), 918-924), it appears that environmental allergens are associated with both the clinical signs and lower airway inflammatory pathology observed in horses with MEA (Bond et al., J Vet Intern Med.2018, 52(6), 2088-2098). Fettelschoss-Gabriel et al. (2021) suggest the use of a virus-like particle (VLP)-based therapeutic equine IL-5 vaccine for the treatment ofIBH. The vaccine has been tested in horses with IBH in placebo-controlled randomized double-blind clinical trials and both showed reduction of IBH lesion scores after vaccination throughout the whole IBH season. However, these virus-based approaches have the disadvantage that the immune reaction may be triggered excessively due to the polyclonal glycosylated antibodies, which are produced in the horse's body after vaccination. Further there might be the risk of the generation of autoimmune antibodies for life-time as an endogenous IL-5 protein is used for vaccination. Fettelschoss-Gabriel et al. (2021) explicitly teaches away from the use of therapeutic antibodies in horses. It is stated on page 7, right column, end of second paragraph: "However, the size and weight of a horse currently precludes usage of monoclonal antibodies in this species due to excessive costs of such a therapy based on passive vaccination requiring frequent injection of large amounts of antibody, which are generally dosed per kg of patient weight. Hence, the benefits of cytokine-blocking antibodies used in humans can be translated to large companion animals, such as horses, by active vaccination." While polyclonal, serum-purified diagnostic antibodies against equine IL-5 are commercially available, they may not be used as a medicament since they do not contain at least partially an equine backbone, meaning that the constant regions of the diagnostic antibodies are not at least partially equine. "At least partially" means a part but, maybe, not the whole entity to which the term is referring to. By indicating "at least", everything between a part and the whole entity is comprised. In connection with the equine backbone, it means that at least a part up to the complete amino acid sequence of the antibody backbone is of equine origin or has a high sequence homology of at least 95% to the respective equine amino acid sequence. The backbone typically comprises the constant parts, i.e., the constant region of the heavy and of the light chain, of an antibody. Thus, "at least partially equine" with respect to the equine backbone comprises the full constant regions of the heavy and/or light chain of the antibody as well as parts thereof, like tmncated forms. It also includes that parts of the backbone may be derived from an organism other than horse. It is, however, preferred that the whole backbone is equine, i.e., is of equine origin. More preferably, the backbone is of equine origin but glycosylation sites are removed by amino acid exchange or chemical modification so that no glycosyl residue may be attached. In contrast thereto, the above-described diagnostic antibodies of the prior art are mainly derived from goat or rabbit and may not be used as a medicament since they would trigger an immune response from the horse's immune system against the antibodies of the foreign species. The prior art antibodies are not at least partially equine and do not comprise at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof. The diagnostic antibodies also comprise the naturally occurring glycosylation sites. Hence, an efficient treatment of horses suffering from allergic conditions is not available to date. Contrary to the suggestions in the art, the inventors have developed antibodies and antibody derivatives specifically binding to IL-5 in horses. Those antibodies show high binding activity and high neutralization efficiency in in vitro studies. Further, they have also been able to show a high safety and tolerability in target animal studies that support the effectiveness of the antibodies in the treatment of allergic conditions in horse, in particular, in the treatment of IBH. Due to their ability to neutralize IL-5 activity in allergic conditions in horses, they offer an effective and specific treatment for horses suffering from allergic conditions. The present invention provides an isolated antibody or antibody derivative that specifically binds to equine IL-5 and comprises at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof. The presence of at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof diminishes the activation of the horse's immune system against the administered antibodies or antibody derivatives. Thereby, the effectiveness of the antibody or antibody derivative therapy is increased. The present invention explicitly and preferably provides aglycosylated antibodies and antibody derivatives, which, in contrast to the VLP-based vaccine, explicitly avoid triggering the immune system of the horse, i.e., does not lead to a further enhancement of the immime reaction of the horse. To further ensure highly reduced interaction with the endogenous immune system, the IgG subclass, more preferably, the IgG6 subclass, may be used. This subclass is known to not interact with cells of the innate and adaptive immune system. Thereby, an over-reaction of the horse to the antibody or antibody derivative of the invention as well as the generation of autoimmune reactions is prevented. Moreover, antibodies and antibody derivatives of the IgG and IgG6 only have a blood half-life of about 21 days. This allows for a very controlled and specific administration of the antibody or antibody derivative and minimizes side effects for the horse. The basic vaccination regimen with the VLP-based vaccine consists of three vaccinations in the first year and an annual booster for the following years. The was higher in the second treatment and paralleled by more sustained antibody. Vaccinated horses had no IL-5-specific antibodies shortly before the booster in the second and the third year, indicating the reversibility of vaccine-induced anti-IL-5 antibodies. However, whether repeated vaccination over a period of years may eventually permanently change the immune system cannot be inferred from the study available so far. The advantage of monoclonal antibodies is that they do not influence the immune system itself, but directly capture IL-5 (Fettelschoss-Gabriel et al.. Allergy 2019, 74(3), 572-582. As IBH is an oven-eaction of the endogenous immune system, the prevention of a further enhancement of this immune reaction makes the antibody or antibody derivative superior to the VLP-based therapeutic vaccines of the prior art. As mentioned above, for the discussed allergic conditions, an effective treatment is still lacking. In addition, also diagnostic methods with high sensitivity are needed and the antibodies and antibody derivatives of the invention provide for a specific diagnostic tool to target IL-5 activity in horses, which can be used as a marker for allergic conditions. Thus, it is a purpose of the present invention to provide for an efficient treatment of allergic conditions in horses and a diagnostic tool to determine an allergic condition in a horse. For this purpose, the present invention provides an isolated antibody or antibody derivative that specifically binds to equine IL-5 comprising at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof. Preferably, the isolated antibody or antibody derivative comprises at least one equine constant light chain or fragments thereof and/or at least one equine constant heavy chain or fragments thereof, optionally, as depicted in SEQ ID NO: 56 (constant region light chain amino acid sequence), SEQ ID NO: 57 (constant region heavy chain amino acid sequence) or SEQ NO: 151 (amino acid sequence CH1 constant region of heavy chain), respectively, or having at least 75%, at least 80%, at least 90%, preferably, at least 95% or 97%, more preferably 98% and, particularly preferred, 99% sequence homology thereto. Also fragments of SEQ ID NO: 56, 57 and/or 151 may be used. In another preferred embodiment, the isolated antibody or antibody derivative comprises at least two equine light chain(s) and at least two equine constant heavy chain(s) or fragments thereof, optionally as depicted in SEQ ID NO: 56 (constant region light chain amino acid sequence), SEQ ID NO: 57 (constant region heavy chain amino acid sequence) or SEQ ID NO: 151 (amino acid sequence CH1 constant region of heavy chain), respectively, or having at least 75%, at least 80%, at least 90%, preferably, at least or 97%, more preferably 98% and, particularly preferred, 99% sequence homology thereto. More preferably, the isolated antibody or antibody derivative is a monoclonal antibody or antibody derivative. In addition, and particularly preferred, the (monoclonal) isolated antibody or antibody derivative is aglycosylated. Preferably, the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 1,155, 156,157,158,2,3, 4, 5, 6, 7, 8, 9, 10, 11,12,13, 14, 15, 16,17,18, 19, 20 and 21. Also preferred is that the isolated antibody or antibody derivative comprises at least one CDR amino acid sequence having at least 99% sequence homo logy to any of the sequences as depicted in any one ofSEQ ID NOs: 1,155,156,157,158,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and21. More preferably, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155,156, 157 and 158, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3, 4 and 5, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6, 7, 8 and 9, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 10, 11, 12, 13 and 14, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQIDNOs: 15, 16, 17, 18, 19, 20 and 21. Even more preferably, the isolated antibody or antibody derivative according to the invention comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23, 24, 25, 26, 27, 28,29,30,31, 32, 33, 35, 36 or 37, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39, 40,41,42,43,45, 49,50,51,52 or 53. In another preferred embodiment of the invention, the isolated antibody or antibody derivative according to the invention comprises a light chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 54, and/or comprises a heavy chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 55 or 153, more preferably as in SEQ ID NO: 55. In another preferred embodiment, the isolated antibody or antibody derivative is an isolated IgG antibody or antibody derivative. More preferably, the isolated antibody or antibody derivative is an isolated IgG6 antibody or antibody derivative. Optionally, the isolated antibody or antibody derivative according to the invention has a neutralizing ability towards IL-5, in particular, equine IL-5. Preferably, the antibody does not bind to bovine serum albumin (BSA) or only to a very minor extent. Further optionally, the isolated antibody or antibody derivative according to the invention is equinized. The present invention also provides an isolated nucleic acid molecule comprising at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention. Further provided is a vector comprising at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention. In a preferred embodiment, the vector is an expression vector. Also provided is a host cell comprising the vector of the invention or at least one nucleic acid sequence encoding the isolated antibody or antibody derivative according to the invention and as described herein. Preferably, the host cell is a eukaryotic host cell. In another embodiment, the invention provides a pharmaceutical composition comprising the isolated antibody or antibody derivative according to the invention and a pharmaceutically acceptable excipient. In one embodiment, the isolated antibody or antibody derivative or the pharmaceutical composition according to the invention and as described herein are for use as a medicament. Preferably, the isolated antibody or antibody derivative or the pharmaceutical composition of the invention are provided for use in the treatment of an allergic condition of a horse. More preferably, the allergic condition is selected from a group consisting of equine allergic skin disease, equine allergic respiratory disease and inflammatory disease. In another preferred embodiment, the equine allergic skin disease is selected from a group consisting of IBH, atopic dermatitis, food hypersensitivity, allergic and irritant contact dermatitis and urticaria, more preferably, the equine allergic skin disease is IBH or atopic dermatitis, most preferably, the equine allergic skin disease is IBH. The equine allergic respiratory disease is, preferably, equine asthma. It is also preferred that the allergic condition is IBH or equine asthma. Moreover, provided is an in vitro method for the diagnosis of an allergic condition in a horse comprising contacting the isolated antibody or antibody derivative according to the invention with a biological sample, preferably obtained from a horse, which might be suspected of suffering from an allergic condition. In another aspect of the invention, a method of manufacturing the isolated antibody or antibody derivative according to the invention is provided, comprising expressing the isolated antibody or antibody derivative in a host cell. Preferably, the isolated antibody or antibody derivative is expressed in a host cell according to the invention, such as a eukaryotic cell. The term "antibody" denotes an intact immunoglobulin having two light and two heavy chains. An isolated antibody or antibody derivative may be a polyclonal antibody, a monoclonal antibody, a synthetic or artificial antibody, a recombinant antibody, a bi-, tri- or multi-specific antibody, a chimeric antibody and/or an equinized antibody. Also combinations thereof are possible, e.g., the antibody may be a monoclonal and equinized antibody. An "anti-IL5" or "a-IL5" antibody is an antibody, which binds to IL-5, in particular to equine IL-5, as the antibody of the present invention. Preferably, the antibody of the invention specifically binds to equine IL-5. The antibody of the present invention is further defined in the claims. Presently, there are five major classes ofimmunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes) (Wagner, Dev Comp Immunol.2006, J0(l-2), 155-64). Seven IgG subclasses exist in the horse. For the antibody and antibody derivative of the present invention, the subclass IgG6 is preferred due to its lowest effector function. IgG has the advantage of a half-life of about 21 days in the horse. In the case of eventually occurring side effects, the antibodies will not remain in the horse's body for a long time and side effects may be controlled effectively. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 8, s, y, and [i, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known in multiple species. The prevalence of individual isotypes and functional activities associated with these constant domains are species- specific. The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (K) and lambda (X), based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. An ,,intact immunoglobulin" or "native immunoglobulin" of the IgG subclass is usually a heterotetrameric glycoprotein having a molecular weight of about 150,000 Daltons, composed of two identical light (L) chains and two identical heavy chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH, heavy chain variable region) followed by a number of constant domains (heavy chain region). Each light chain has a variable domain at one end (VL, light chain variable region) and a constant domain at its other end (light chain constant region); the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains (regions). The variable domains (regions) of the isolated antibody and antibody derivative of the present invention bind to the antigen, i.e., to equine IL-5. As stated above, in contrast to the usual appearance of a native immunoglobulin of the IgG subclass, the isolated antibody or antibody derivative of the present invention is, preferably, aglycosylated. The term "antibody derivative" includes all short or abbreviated forms of the antibody described herein as well as antibody fragments, i.e., to non-intact immunoglobulins. An antibody derivative includes the antigen binding portion of the antibody, which binds to, and preferably neutralizes, specifically equine IL-5. The antibody derivative may be truncated in the Fc region of the antibody or may not contain an Fc region at all. The term includes isolated single (heavy or light) antibody chains, an Fv construct, F(ab')2 fragment, a Fab construct, an Fc construct and a light or heavy chain variable or complementarity determining region (CDR) sequence. Moreover, the term "antibody derivative" comprises antibody conjugates, wherein the antibody is conjugated to another functional molecular entity, optionally via a linker, such as a short peptide (1-10 amino acids) or another organic linker. Conjugated denotes a covalent bond between the antibody and another molecular entity, optionally, an organic or inorganic molecular moiety, which may equip the antibody with additional properties and/or functionalities. This may include at least a moiety to facilitate purification, detection, increase bioavailability and/or biocompatibility. Such a molecular entity may also be used to localize the antibody and antibody derivative to a desired region within the horse's body. The linker may be cleavable by chemical means or by applying heat, light or other. A fragment of equine constant light chain or equine constant heavy chain includes any truncated, i.e., shortened, form of an equine constant light chain or equine constant heavy chain and may contain only few (at least 10 amino acids). The term "hypervariable region", as used herein, refers to the amino acid residues of an antibody or antibody derivative, which are responsible for antigen binding, e.g., bind to equine IL-5. The hypervariable region comprises amino acid residues from a "complementarity determining region" (CDR) and/or those residues from a "hypervariable loop". "Framework" (FR) residues are those variable domain residues other than the hypervariable region residues. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" (Fragment antigen binding) fragments, each with a single antigen-binding site, and a residual "Fc" fragment, named after its ability to crystallize. Pepsin treatment of antibodies yields an F(ab')2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen. "Fv" is the minimum antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. It is in this configuration that the three hypervariable regions of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six hypervariable regions confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three hypervariable regions specific for an antigen) can recognize and bind antigen with neutralizing ability, although at a lower binding activity than the entire binding site. The Fab also contains the constant domain of the light chain and the first constant domain of the heavy chain (CH1). Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain including one or more cysteine(s) from the antibody hinge region. Fab'-SH is the designation for Fab\ in which the cysteine residue(s) of the constant domains bear a thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments, which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known. The term "specifically", in the context of antibody or antibody derivative binding, refers to high avidity and/or high binding activity of an antibody/derivative to a antigen, i.e., a polypeptide, or epitope. The binding activity of a protein- protein interaction can be described as the strength of the interaction between a receptor and its respective ligand. An example of this is an antibody's or antibody derivative's epitope and an antigen (here, in particular, IL-5). In this interaction, the antigen binds to the epitope through a variety of different types of bonds, such as hydrogen bonds, ionic bonds, van der Waals interactions and electrostatic forces. The term "specifically" also includes that the antibody or antibody derivative binds to a far greater extent to equine IL-5 than to other targets, including BSA. The term "isolated" means that the antibody, antibody derivative or nucleic acid molecule is separated and/or recovered from a component of a natural environment. Contaminant components of a natural environment are materials that would interfere with diagnostic or therapeutic uses for the material, and may include enzymes, and other proteinaceous solutes. With respect to nucleic acid, an isolated nucleic acid may include one that is separated from the 5' to 3' sequences, with which it is normally associated in the chromosome. In preferred embodiments, the material will be purified to greater than 95% by weight of the material, and most preferably more than 99% by weight. Isolated material includes the material in situ within recombinant cells since at least one component of the material's natural environment will not be present. Ordinarily, however, isolated material will be prepared by at least one purification step. "IL-5" (interleukine-5) is the most potent activator of eosinophils and is produced by Th2 cells and ILC2s in mammals. IL-5 is also reported to have a role in eosinophil extracellular trap cell death and binds to the IL-5 receptor. It is further involved in B- cell growth and immunoglobulin secretion. Moreover, IL-5 was also shown to be upregulated in other allergic horses and, in particular, in horses with asthma. The antibody and antibody derivative of the present invention specifically binds to an equine IL-5, i.e., an IL-5 from horse. A "monoclonal antibody or antibody derivative" as defined herein is a single antibody or antibody derivative that is generated by hybridoma technology or that is selected by a phage display approach and cloned into an immunoglobulin, preferably equine IgG backbone (i.e., constant regions are equine and belong to the IgG or IgG6 immunoglobulin class). This construct may then be transferred on a DNA basis into single-cloned eukaryotic cells for production. A "chimeric" antibody (immunoglobulin) is an antibody, in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or artificially created, as well as fragments of such antibodies, as long as they exhibit the desired biological activity. Typically, chimeric antibodies are antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from antibody variable and constant region genes belonging to different species or are artificially designed. For example, the variable segments of the genes from a non-equine monoclonal antibody may be joined to equine constant segments. In this embodiment, the variable domains including the antigen binding site is derived from a human antibody library while the constant domains are of equine origin. In one embodiment, the isolated antibody or antibody derivative of the present invention is an isolated chimeric antibody or antibody derivative, preferably, an isolated chimeric antibody or antibody derivative comprising at least one member selected from a group consisting of a non-equine light chain variable region, non- heavy chain variable region, at least one non-equine CDR and combinations thereof. "None-equine" includes all organisms except horses, i.e., animals, which belong to the family ofEquidae. Examples of none-equine is human. "Equine" in the sense of the present invention is to be understood as "derived originating from horse" or "of horse origin". The antibodies and antibodies derivatives of the present invention also bind to IL-5 from donkeys, hinnies and mules. "Homologous" or having a certain percentage of sequence homology with respect to amino acid or nucleic acid sequences means that the sequences or antibody chains share at least 80% sequence identity, preferably at least 85% or at least 90%, more preferably at least 95% or 97% and even more preferably at least 98% or 99% sequence or homology. Most preferred is a sequence identity of 100%. In this case, the comprised amino acid or nucleic acid sequence is 100% identical to the respective SEQ ID NO. Hence, "being identical to" means 100% sequence identity or homology. "Identity" or "homology" with respect to an amino acid or nucleic acid sequence is defined herein as the percentage ofamino acid or nucleic acid residues in the differing sequence that are identical with the parent or original amino acid or nucleic acid residues, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. None of N-terminal, C-terminal, or internal extensions, deletions, or insertions into the antibody sequence shall be construed as affecting sequence identity or homology. The sequence identity or homology may be determined by standard computational tools available to the skilled person, such as BLAST or similar. "Equinized" forms ofnon-equine (e.g., marine) antibodies are genetically engineered antibodies that contain partly sequences derived from non-equine immunoglobulin. Equinized antibodies are equine immunoglobulin sequences (recipient antibody) in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-equine species (donor antibody), such as human or artificial, having the desired specificity, binding activity and capacity. In some instances, framework region (FR) residues of the equine immunoglobulin sequences are replaced by corresponding non-equine residues. Furthermore, equinized antibodies include residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the equinized antibody will include substantially all or at least one, and typically two, variable domains, in which all or substantially all of the hypervariable regions correspond to those of a non-equine immunoglobulin sequence and all or all of the FRs are those of an equine immunoglobulin sequence. The equinized antibody also will comprise a complete or at least a portion of an immunoglobulin constant region (Fc), typically that of an equine immunoglobulin sequence. The term "neutralizing ability towards IL-5" means that the antibody or antibody reaches the IC5o value at a molar ratio of antibody to monomeric antigen in the range of 0.3-1. The IC50 value, which is the antibody or antibody derivative concentration needed to reduce the binding to 50% of equine IL-5 (eqIL-5) to its receptor, when preincubated with the antigen and then added to Expi293F cells which present the eqIL-5 receptor on their surface. Binding of the antigen to the cells is detected in the flow cytometer by fluorescent labelling via the His-tag and a fluorescently labelled antibody. To obtain the relative binding, the measured fluorescence signal (median) is to be related to the signal when only antigen and cells, but no inhibiting antibody, were present. 66 "Aglycosylated" denotes that the antibody or antibody derivative is not glycosylated due to a mutation in the constant region of the heavy chain, when compared to the native equine constant region heavy chain amino acid sequence. While aglycosylated antibodies or antibody derivatives are equal in terms of antigen binding, pharmacokinetics, and biodistribution, they have the advantage to bypass the drawbacks of glycosylated antibodies that include glycan heterogeneity and requirement of high capital investment for biomanufacturing. For example, glycosylated antibodies or glycosylated antibody derivatives may have an unpleasant role in recmiting innate immimity effector cells. This disadvantage is overcome since the antibodies and antibody derivatives of the present invention are preferably aglycosylated, i.e., are not glycosylated. terms "nucleic acid", "polynucleotide", "nucleic acid molecule" and the like may be used interchangeably and refer to a series of nucleotide bases (also denoted as "nucleotides") in DNA and RNA. The nucleic acid molecule of the invention may contain deoxyribonucleotides, ribonucleotides, and/or their natural or artificial analogs. The term "nucleic acid molecule" includes single-stranded and double- molecules. A nucleic acid molecule can be a gene or gene fragment, exons, introns, a DNA molecule, like cDNA, an RNA molecule, like mRNA, recombinant nucleic acid molecules, plasmids, and other vectors, primers and probes. Both 5' to 3' (sense) and 3' to 5' (antisense) polynucleotides are included. The nucleic acid molecule of the present invention is preferably isolated as defined hereinabove. " (; E (;- ncoding at least partially" means that a nucleic acid sequence may encode the full amino acid sequence of the heavy and/or light chain or may encode only parts of the amino acid sequences of the heavy and/or light chain of the antibody or antibody derivatives of the invention, i.e., only the heavy chain or only the light chain, only the variable region of the heavy or light chain or only the constant region of the heavy or light chain. Different combinations are possible and also only parts of the heavy and light chains may be encoded by a single nucleic acid sequence or single nucleic acid molecule. For producing the isolated antibody or antibody derivative, different host cells, (isolated) nucleic acid molecules or vectors may be used. For example, the heavy and light chains of the antibody and antibody derivative may be produced separately in any order or in parallel using different host cells, isolated nucleic acid molecules and/or vectors. In the case of antibody derivatives, not the full amino acid sequence of the heavy and/or light chain may be needed and the nucleic acid sequences disclosed herein may be used in truncated form, i.e., not in the full length as described herein,to produce e.g., truncated antibody derivatives. Hence, according to the needs of the practitioner, the amino acid and nucleic acid sequences described herein may be truncated. The term "host cell" refers to a prokaryotic or eukaryotic cell (e.g., bacterial cells or mammalian cells) whether located in vitro or in vivo. For example, host cells may be located in a transgenic animal. Host cell can be used as a recipient for vectors and may any transformable organism that is capable of replicating a vector and/or expressing a heterologous nucleic acid encoded by a vector. Thereby, the antibody or antibody derivative of the present invention may be produced in large quantities and in reproducible quality. The skilled person is well-aware of standard techniques of protein and in particular antibody or antibody derivative expression as well as for protein purification. In this context, "protein" includes the antibody and antibody derivative of the present invention. The (expression) vector carrying the nucleic acid sequence encoding the antibody or antibody derivative according to the present invention can be introduced into an appropriate host cell by any of a variety of suitable means, including transformation, transfection, conjugation, protoplast fusion, calcium phosphate-precipitation and application with polycations such as diethylaminoethyl (DEAE) dextran, and such mechanical means as electroporation, direct microinjection, and microprojectile bombardment. The term "allergic condition" is defined herein as a disorder or disease caused by an interaction between the immune system and a substance foreign to the body, i.e., the allergen. Often, the reaction of the body to the foreign substance is too strong so that it poses a problem to the subject, which got in contact with the allergen. Preferably, the allergic condition is IBH or equine asthma. Most preferably, the allergic condition is IBH. The term includes allergic skin diseases, allergic respiratory diseases and inflammatory diseases. The term "equine allergic skin disease" includes diseases that manifest with symptoms correlated to the skin of the horse, such as IBH, atopic dermatitis, food hypersensitivity, allergic and irritant contact dermatitis and urticaria, preferably, the equine allergic skin disease is IBH or atopic dermatitis, most preferably, the equine allergic skin disease is IBH. Also combinations thereof are possible. The terms "insect-bite hypersensitivity" (IBH), "Culicoides hypersensitivity" (CH), "sweet itch", "kasen", "Queensland itch", "summer dermatitis", "equine dermatitis" and "summer eczema" denote allergic reactions to bites from mosquitos and midges (Culicoides) but also to other insects (percutaneously). These terms may be used interchangeably herein. The disease is characterized by pruritus and secondary lesions ofalopecia and crusting. The distribution on the horse's body depends on the species of Culicoides feeding on the horse. The classic distribution is the dorsally distributed disease with lesions found on the face, mane, withers, rump, and tail (mainly denoted as sweet itch). Ventrally feeding Culicoides cause lesions in the intermandibular space and on the ventral body wall. Combinations of both are possible, too. As an effective therapy is still missing to date, minimizing the exposure to the insects is the only option for horse owners. For acute cases, glucocorticoids with many adverse effects are the only therapy available but not suitable as durable therapy. Further, it is helpful to stable the affected horses from dusk till dawn as well as to use insect repellents, fans in the stalls, fly masks, and fly sheets. In general, the diagnosis of IBH is made by clinical history, clinical signs, improvement due to insect control and intradermal and/or ELISA testing (serology-based) (Fadok, Vet. Clin. Equine 2013, 29, 541-550). "Atopic dermatitis" is a pmritic inflammatory disease of the skin in mammals, mainly caused by contact with diverse antigens (e.g., pollen, mold, food, mites and dust, both inhaled and percutaneously) that can be followed by an inflammatory respiratory disease later in disease progress (Misery and Stander, edt.2010. Pruritus. London, Springer, Marsella, VetSci.2021, 8(7), 124.). Clinical signs of equine atopic dermatitis are pruritus and skin inflammation that typically affects the face, ears, and glabrous areas. Many atopic horses are also insect allergic. "Food Hypersensitivity" is all allergic condition caused by food allergens, like grains, grasses, supplements or food additives. "Allergic and irritant contact dermatitis" including allergic eczema is an allergic condition that affects the skin of the horse and other mammals and results in a disrupted epidermis associated with an itchy inflammatory response due to allergic sensitization. Allergens include lotions, shampoos etc., nickel, copper, zinc (percutaneously). Allergic contact dermatitis can either occur as a single event or chronically (Nedorost, Dermatologic Clinics 2020, J5(3) 301-308). "Urticaria" (including "hives") is a skin condition characterized by the formation of itchy red or whitish raised patches, usually caused by an allergen (Misery and Stander Pruritus 2010, London, Springer). An "equine allergic respiratory disease" is an allergic condition of the horse, which manifests in symptoms of the respiratory system of the horse. An "inflammatory disease" is a condition characterized by inflammation. The term "equine asthma" (according to ACVIM Consensus Statement 2016) includes all respiratory diseases from inflammatory airway diseases (IAD) with medium to high severity, heaves and Recurrent Airway Obstructions (RAO). RAO was formerly known as chronic obstruction pulmonary disease (COPD) and, in the German- speaking area, it is still called chronic obstmctive bronchitis (COB). COB is a chronic, inflammatory and non-infectious disease of the lung. Hence, the term "equine asthma" includes IAD, heaves, RAO, COPD and COB. A "pharmaceutical composition" in the sense of the invention denotes a combination of active agent, here the antibody or antibody derivative of the invention, and another compound or composition, which can be inert (like water or buffer), or active, such as an adjuvant. A "therapeutically effective amount" (or "effective amount") refers to an amount of an active ingredient, i.e., the antibody or antibody derivative according to the invention, sufficient to effect beneficial or desired results, when administered to a subject or patient, which is in the present case, the horse affected by the allergic condition. An effective amount can be administered by one or more routes of administration, applications or dosages. Administration may be weekly, every second or third week, once per month, every second, third or six month or yearly depending on the decision of the veterinary based on the disease state and condition of the affected horse. Administration may by any route of administration (see below). A therapeutically effective amount of a composition according to the invention may be readily determined by one of ordinary skill in the art or the horse owner if trained by the veterinary. In the context of this invention, a "therapeutically effective amount" is one that produces an objectively measurable change in one or more parameters associated with treatment of the allergic condition, including clinical improvement in symptoms or the prevention of worsening of symptoms. Of course, the therapeutically effective amount will vary depending upon the particular horse/subject and condition being treated, the race, gender, weight and age of the horse, the severity of the disease condition, the time point of the year in case of seasonal allergies, the particular compound and composition chosen, stabling of the horse, potential additional medicaments taken by the horse, the feeding, the dosing regimen to be followed and the timing of administration. "Treatment", "treating", and the like, refers to both therapeutic treatment and prophylactic or preventative measures. Animals in need of treatment include those already with the allergic condition as well as those, in which the allergic condition is to be prevented, such as horses, which are suspected to have a predisposition to develop an allergic condition due to, e.g., pedigree or race. The term "treatment" or "treating" of an allergic condition includes preventing or protecting against the allergic condition (that is causing the clinical symptoms not to develop); inhibiting the allergic condition (i.e., arresting or suppressing the development of clinical symptoms, and/or relieving the allergic condition (i.e., causing the regression of clinical symptoms). As will be appreciated, it is not always possible to distinguish between "preventing" and "suppressing" an allergic condition since the ultimate inductive event or events may be unknown or latent. Accordingly, the term "prophylaxis" will be understood to constitute a type of "treatment" that encompasses both "preventing" and "suppressing". The term "treatment", thus, may include "prophylaxis". The antibodies and antibody derivatives according to the invention provide for a specific diagnostic tool to target IL-5 activity in horses, which can be used as a marker for allergic conditions. As expressed in Fettelschoss-Gabriel et al., 2021, diagnostic tools with high sensitivity and specificity need to be developed. The answer to this need is presented by the invention, which provides an isolated antibody or antibody derivative that specifically binds to an equine IL-5. Preferably, the isolated antibody or antibody derivative are monoclonal, chimeric and/or equinized. The antibodies and antibody derivatives of the present invention may be produced by standard means in standard expression hosts using standard expression vectors. Information can e.g., be found in "Recombinant Protein Expression in Mammalian Cells" (2018, Springer, Ed. D.L. Hacker, MIMB, vol.1850), "Textbook on Cloning, Expression and Purification ofRecombinant Proteins" (2022,1s. ed., Ed. Kakoli Bose, Springer) and "Gene Expression Systems - Using Nature for the Art of Expression" (1998, Academic Press, 1 ed., Ed. J. Fernandez, J. Hoeffler). Typical expression hosts comprise bacterial cells or eukaryotic cells, preferably the expression cell or host cell is a eukaryotic cell. Cloning techniques are also well known to the skilled person. The pharmaceutical composition provided with the present invention comprise the antibody or antibody derivative of the present invention and a pharmaceutically acceptable excipient. The antibody or antibody derivative is to be administered in a therapeutically effective amount. The amount or concentration within the pharmaceutical composition depends on the stability that can provided by the pharmaceutical composition to the antibody or antibody derivative. The stability must be suitable enough to ensure a storage for a convenient period of time,;". e., degradation must be in a regulatory accepted range. Pharmaceutically acceptable excipients suitable for use in the present invention are well known to those of skill in the art. Such carriers include, without limitation, water, saline, in particular, normal saline (i.e., a solution of 0.90% w/v ofNaCl), buffered saline, phosphate buffer, phosphate buffered saline, alcoholic and aqueous solutions, emulsions or suspensions. Other conventionally employed diluents, adjuvants and excipients, may be added in accordance with conventional techniques. Such carriers can include ethanol, polyols, and suitable mixtures thereof, vegetable oils and injectable organic esters. Buffers and pH adjusting agents may also be employed. Buffers include salts prepared from an organic acid or base. Representative buffers are organic acid salts, such as salts of citric acid, like citrates, ascorbic acid, gluconic acid, histidine-HCl, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid, (hydroxymethyl)aminomethane (Tris) or phosphate buffers. Parenteral carriers can include sodium chloride solution, Ringer's dextrose, dextrose, trehalose, sucrose, and sodium chloride, lactated Ringer's or fixed oils. Intravenous carriers comprise fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose and the like. Preservatives and other additives such as, for example, antioxidants, chelating agents (e.g., EDTA), inert gases and the like may also be provided in the pharmaceutical carriers. The preparation of these pharmaceutically acceptable compositions, from the above-described components, having appropriate pH isotonicity, stability and other conventional characteristics is within the skill of the art. a preferred embodiment, the pharmaceutically acceptable excipient is phosphate buffered saline (PBS). The concentration of the antibody in the pharmaceutical composition is typically in the range of 0.1 to 20 mg/mL, more preferably in the range of 2 to 10 mg/mL and even more preferably in the range of 5 to 10 mg/mL. The pharmaceutical compositions according to the invention may be administered via any possible route of administration, such as via the parenteral, oral, subcutaneous, transdermal route, inhalational or pulmonary route, nasal, sublingual/buccal, mucosal, topical, rectal and/or intravenous route. Preferably, the pharmaceutical composition of the present invention is administered via the subcutaneous route of administration. The present invention also provides for an in vitro method for the diagnosis of an allergic condition in a horse. Detecting IL-5 in companion animals known to be or suspected of having a pruritic and/or allergic condition. By contacting the antibody or antibody derivative of the invention with a biological, preferably liquid, sample obtained from a horse, which may be a fluid obtainable from the body, including a blood or serum sample, tissue sample or else, the presence ofIL-5 may be determined. Via comparison with a reference sample, e.g., obtained from a non-allergic horse, an increased level of IL-5 may be determined. Contacting may be performed by mixing. An increased level or concentration of IL-5 may then be indicative of an allergic condition of a horse, optionally in combination with a further examination by a veterinary. Methods for detecting and comparing antibody or antibody derivative concentrations in a sample include enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunospot (ELISPOT), Western blot (WB), immunoprecipitation (IP) and others and are well-known to the person of ordinary skill in the art. Information can e.g., be found in "Molekularbiologische Methoden 2.0" (2018, utb, 2nd ed., Thomas Reinard). Particularly useful are immunoassays like ELISA, which detect or quantitate IL-5 or anti-IL-5 antibodies in a sample obtained from a horse. For this purpose, the antibody or antibody derivative of the present invention may be covalently or non-covalently bound to a detectable molecular entity (such as a polypeptide or small molecule), which includes a fluorescent, radioactive or otherwise detectible molecular entity. An immunoassay for IL-5 typically comprises incubating a clinical or biological sample of the horse in the presence of a detectably labeled antibody or antibody derivative of the present invention capable of selectively binding to IL-5, and detecting the antibody or antibody derivative, which is bound in a sample by standard means depending on the used detectable molecular entity. The biological sample may be any body fluid, including serum, plasma, tissue and cell lysate, saliva, sputum and bronchoalveolar lavage. Amino acid and nucleic acid sequences: SEQ ID NO: 1 Light chain CDR 1 (NOL1-13): NIGSKS. SEQ ID NO: 1 is the most preferred light chain CDR1 amino acid sequence. SEQ ID NO: 2 Light chain CDR 3 (NOL9-13): QVWDSSSDPVV. SEQ ID NO: 2 is the most preferred light chain CDR3 amino acid sequence. SEQ ID NO: 3 Light chain CDR 3 (NOL1-8): QVWDSGDGX2PRV, wherein X2 can be H or N. SEQ ID NO: 4 Light chain CDR 3 (NOLI, 2, 3, 4, 6, 8): QVWDSGDGHPRV. SEQ ID NO: 5 Light chain CDR 3 (NOL5 and NOL7): QVWDSGDGNPRV. SEQ ID NO: 6 Heavy chain CDR 1 (NOL1-13): GFTFXsSYG, wherein Xs can be S, G orR, preferably Xs is S. SEQ ID NO: 7 Heavy chain CDR 1 (NOL1-11): GFTFSSYG. SEQ ID NO: 7 is the most preferred heavy chain CDR1 amino acid sequence. SEQ ID NO: 8 Heavy chain CDR 1 (NOL 12): GFTFGSYG. SEQ ID NO: 9 Heavy chain CDR 1 (NOLI 3): GFTFRSYG. SEQ ID NO: 10 Heavy chain CDR 2 (NOL1-13): IX4X5DGX6NK, wherein X4 can be S or W, X5 can be Y or N and X6 can be S or R, preferably X4isS,X5isYandX6isS. SEQ ID NO: 11 Heavy chain CDR 2 (NOL9-12): ISYDGSNK. SEQ ID NO: 11 is the most preferred heavy chain CDR2 amino acid sequence. SEQ ID NO: 12 Heavy chain CDR 2 (NOL13): ISNDGSNK. SEQ ID NO: 13 Heavy chain CDR 2 (NOLI, 3, 4, 5, 6, 7, 8): IWYDGSNK. SEQ ID NO: 14 Heavy chain CDR 2 (NOL2): IWYDGRNK. SEQ ID NO: 15 Heavy chain CDR 3 (NOL9-13): X7KGXsKIQLWFAAFDI, wherein X? can be V or A and Xs can be F or L, preferably X? is V and Xg is L. SEQ ID NO: 16 Heavy chain CDR 3 (NOLI I): VKGLKIQLWFAAFDI. SEQ ID NO: 16 is the most preferred heavy chain CDR3 amino acid sequence. SEQ ID NO: 17 Heavy chain CDR 3 (NOL9, 12, 13): AKGFKIQLWFAAFDI. SEQ ID NO: 18 Heavy chain CDR 3 (NOLI O): VKGFKIQLWFAAFDI. SEQ ID NO: 19 Heavy chain CDR 3 (NOLI-8): ARGLX9GRSYFDY, wherein X9 can be F or Y. SEQ ID NO: 20 Heavy chain CDR 3 (NOLI, NOL3-7): ARGLYGRSYFDY. SEQ ID NO: 21 Heavy chain CDR 3 (NOL2 and NOL8): ARGLFGRSYFDY. SEQ ID NO: 22 Variable region light chain - full sequence (NOLI-13): X7iX72X73LTQPPSX74SVAPGQTAX75lX76CX77GX7sNIGSKSVHWYX79Q X8oPGQAPVLVVXsiDDX82DRPSGIX83X84RX85SGSNX86GNTAX87LTIX88 RVEX89GDEADYYCQVWDSX9oX9lX92X93X94X95X96X97GGX98X99XlOO XloiXl02Xl03, wherein X7i can be S or Q, X?2 can be Y or S, X?3 can be E or V, X74 can be V, M or I, X?5 can be K or R, X^6 can be A, V or T, Xy? can be E or G, X^s can be D or N, X?9 can be Q or H, Xgo can be R or K, Xsi can be F or Y, X§2 can be T, Aor S, Xs3 can be P or T, Xs4 can be D or E, Xg5 can be F or L, Xg6 can be S or T, Xs^ can be T or A, Xss can be S or G, Xs9 can be V, I, A or S, X9o can be G or S, X9i can be D or S, X92 can be G or D, X93 can be H, N or P, X94 can be P or V, X95 can be R or V, X96 can be V or F, X97 can be F or G, X9g can be G or T, X99 can be T, S or K, Xioo can be K or L, Xioi can be L or T, Xio2 can be T or V and Xio3 can be V or L. SEQ ID NO 23: Variable region light chain - full sequence (NOL9-13): QSVLTQPPSXioSVAPGQTARITCGGNNIGSKSVHWYXiiQKPGQAPVL VVYDDSDRPSGIPERFSGSNXi2GNTATLTIXi3RVEXi4GDEADYYCQV WDSSSDPVVFGGGTKLTVL, wherein Xio can be V or I, Xii can be Q or H, Xi2 can be S or T, Xis can be S or G and Xi4 can be A or S, preferably Xio is V, Xi 1 is Q, Xi2 is S, Xi3 is S and Xi4 is A. SEQ ID NO: 24 Variable region light chain - full sequence (NOLI 1, NOL9): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL SEQ ID NO: 24 is the most preferred variable region light chain full sequence. SEQ ID NO: 25 Variable region light chain - full sequence (NOL 1 -8): SYELTQPPSXi5SVAPGQTAXi6lXi7CEGDNIGSKSVHWYQQRPGQAPV LVVXi8DDXi9DRPSGIX2oDRX2iSGSNSGNTAX22LTISRVEX23GDEADY YCQVWDSGDGX24PRVFGGGX25KLTVL, wherein Xi5 can be M or V, Xi6 can be K or R, Xi7 can be A, V or T, Xi8 can be F or Y, Xi9 can be T or A, X20 can be T or P, X2i can be L or F, X22 can be T or A, X23 can be V or I, X24 can be H or N and X25 can be T or S. SEQ ID NO: 26 Variable region light chain - full sequence (NOLI): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL. SEQ ID NO: 27 Variable region light chain - full sequence (NOL2): SYELTQPPSMSVAPGQTARIACEGDNIGSKSVHWYQQRPGQAPVLVV FDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL. SEQ ID NO: 28 Variable region light chain - full sequence (NOL3): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRLSGSNSGNTAALTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL. SEQ ID NO: 29 Variable region light chain - full sequence (NOL4): SYELTQPPSVSVAPGQTAKIVCEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL. SEQ ID NO: 30 Variable region light chain - full sequence (NOL5): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDADRPSGITDRFSGSNSGNTATLTISRVEIGDEADYYCQVWDSGD GNPRVFGGGTKLTVL. SEQ ID NO: 31 Variable region light chain - full sequence (NOL6): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGSKLTVL. SEQ ID NO: 32 Variable region light chain - full sequence (NOL7): SYELTQPPSVSVAPGQTAKIACEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GNPRVFGGGTKLTVL. SEQ ID NO: 33 Variable region light chain - full sequence (NOL8): SYELTQPPSVSVAPGQTAKITCEGDNIGSKSVHWYQQRPGQAPVLVV YDDTDRPSGIPDRFSGSNSGNTATLTISRVEVGDEADYYCQVWDSGD GHPRVFGGGTKLTVL. SEQ ID NO: 35 Variable region light chain - full sequence (NOL10): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTIGRVESGDEADYYCQVWDSSSD PVVFGGGTKLTVL. SEQ ID NO: 36 Variable region light chain - full sequence (NOL12): QSVLTQPPSISVAPGQTARITCGGNNIGSKSVHWYHQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL. SEQ ID NO: 37 Variable region light chain - full sequence (NOL13): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNTGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVL. SEQ ID NO: 38 Variable region heavy chain - full sequence (NOL1-13): X38VQLX39X4oX4iX42GGVVX43PGRSLRLSCAASGFTFX44SYGMHWVR QAPGKGLEWX45AVIX46X47DGX48NKYYADSVKGRFTISRDNSKNTLY LQX49NSLRAEDTAVYYCX5oX5lGX52X53X54X55X56X57FX58X59X6oX6lX62 X63X64X65X66X6?X68X69X70, wherein Xsg can be E or Q, Xs9 can be Q, E or V, X4o can be Q or E, X4i can be S or T, X42 can be G or E, X43 can be Q or R, X44 can be S, Gor R, X45 can be V or L, X46 can be W or S, X47 can be Y or N, X4§ can be R or S, X49 can be M or I, X5o can be A or V, X5i can be R or K, X52 can be L or F, X53 can be F, Y or K, X54 can be G or I, X55 can be R or Q, X56 can be S or L, X5? can be Y or W, Xss can be D or A, X59 can be Y or A, X6o can be W or F, X6i can be G or D, X62 can be Q or I, X63 can be G or W, X64 can be T or G, X65 can be L or Q, X66 can be V or G, X6? can be T or A, X^s can be V or M, X69 can be S or V and X?o can be S or T. SEQ ID NO: 39 Variable region heavy chain - full sequence (NOL9-13): EVQLX26ETGGGVVX27PGRSLRLSCAASGFTFX2gSYGMHWVRQAPGK GLEWVAVISX29DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCX3oKGX3iKIQLWFAAFDIWGQGX32MVTVSS, wherein Xi6 can be E or V, X2? can be Q or R, X28 can be S, G or R, Xi9 can be Y or N, Xso can be V or A, X3i can be F or L and Xs2 can be T or A, preferably Xi6 is V, X27 is Q, X28 is S, X29 is Y, Xso is V, X3i is L and Xs2 is T. SEQ ID NO: 40 Variable region heavy chain - full sequence (NOLI I): EVQLVETGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCVKGLKIQLWFAAFDIWGQGTMVTVSS. SEQ ID NO: 40 is the most preferred variable region heavy chain full sequence. SEQ ID NO: 41 Variable region heavy chain - full sequence (NOL 1 -8): QVQLQQSXssGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGL EWX34AVIWYDGX35NKYYADSVKGRFTISRDNSKNTLYLQX36NSLRA EDTAVYYCARGLX37GRSYFDYWGQGTLVTVSS, wherein Xss can be E or G, Xs4 can be L or V, Xs5 can be S or R, Xs6 can be M or I and Xs? can be Y or F. SEQ ID NO: 42 Variable region heavy chain - full sequence (NOLI, NOL3, NOL5, NOL6, NOL7): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLYGRSYFDYWGQGTLVTVSS. SEQ ID NO: 43 Variable region heavy chain - full sequence (NOL2): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGRNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLFGRSYFDYWGQGTLVTVSS. SEQ ID NO: 45 Variable region heavy chain - full sequence (NOL4): QVQLQQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQINSLRAEDTAV YYCARGLYGRSYFDYWGQGTLVTVSS. ID NO: 49 Variable region heavy chain - full sequence (NOL8): QVQLQQSEGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WLAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCARGLFGRSYFDYWGQGTLVTVSS. SEQ ID NO: 50 Variable region heavy chain - full sequence (NOL9): WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGTMVTVSS. SEQ ID NO: 51 Variable region heavy chain - full sequence (NOL 10): EVQLVETGGGVVRPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE VYYCVKGFKIQLWFAAFDIWGQGTMVTVSS. SEQ ID NO: 52 Variable region heavy chain - full sequence (NOL 12): EVQLEETGGGVVQPGRSLRLSCAASGFTFGSYGMHWVRQAPGKGLE WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGTMVTVSS. SEQ ID NO: 53 Variable region heavy chain - full sequence (NOL13): EVQLVETGGGVVQPGRSLRLSCAASGFTFRSYGMHWVRQAPGKGLE WVAVISNDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKGFKIQLWFAAFDIWGQGAMVTVSS. SEQ ID NO: 54 Light chain - full sequence (NOLI I): QSVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSD PVVFGGGTKLTVLGPTSTPSVSLFPPSSEELSANKATVVCLISDFSPSGL EVIWKVNDAVTTDGVQTTRSSKQSNGKYAASSYLTRTSAQWKSYSS VSCQVKHQGKTVEKKLSPSECP. SEQ ID NO: 54 is the most preferred light chain full sequence. SEQ ID NO: 55 Heavy chain - full sequence (NOL II): EVQLVETGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCVKGLKIQLWFAAFDIWGQGTMVTVSSASTTAPKVFQLASHSA GTSDSTVALGCLVSSYFPEPVTVSWNSGALTSGVHTFPSVRQSSGLYS LSSMVTVPASSLKSQTYICNVAHPASSTKVDKRIVIKEPCCCPKCPDSK FLGRPSVFIFPPNPKDTLMISRTPEVTCVVVDVSQENPDVKFNWYVDG VEAHTATTKAKEKQDASTYRVVSVLPIQHQDWRRGKEFKCKVNNRA LPAPVERTITKAKGELQDPKVYILAPHREEVTKNTVSVTCLVKDFYPP DINVEWQSNEEPEPEVKYSTTPAQLDGDGSYFLYSKLTVETDRWEQG ESFTCVVMHEAIRHTYRQKSITNFPGK. SEQ ID NO: 55 is the most preferred heavy chain full sequence. SEQ ID NO: 56 Constant region light chain - full sequence: GPTSTPSVSLFPPSSEELSANKATVVCLISDFSPSGLEVIWKVNDAVTT DGVQTTRSSKQSNGKYAASSYLTRTSAQWKSYSSVSCQVKHQGKTV EKKLSPSECP. SEQ ID NO: 57 Constant region heavy chain - full sequence: ASTTAPKVFQLASHSAGTSDSTVALGCLVSSYFPEPVTVSWNSGALTS GVHTFPSVRQSSGLYSLSSMVTVPASSLKSQTYICNVAHPASSTKVDK RIVIKEPCCCPKCPDSKFLGRPSVFIFPPNPKDTLMISRTPEVTCVVVDV SQENPDVKFNWr^/DGVEAHTATTKAKEKQDASTYRVVSVLPIQHQD WRRGKEFKCKVNNRALPAPVERTITKAKGELQDPKVYILAPHREEVT KNTVSVTCLVKDFYPPDINVEWQSNEEPEPEVKYSTTPAQLDGDGSY FLYSKLTVETDRWEQGESFTCVVMHEAIRHTYRQKSITNFPGK. SEQ ID NO: 58 Nucleic acid sequence heavy chain CDR1 NOLI 1, NOLI, NOL3, NOL4, NOL5, NOL6, NOL7, NOL8, NOL9, NOL10: GGATTCACCTTCAGTAGCTATGGC SEQ ID NO: 59 Nucleic acid sequence heavy chain CDR1 NOL2: GGATTCACTTTCAGTAGCTATGGC SEQ ID NO: 69 Nucleic acid sequence heavy chain CDR1 NOL12: GGATTCACCTTCGGTAGCTATGGC SEQ ID NO: 70 Nucleic acid sequence heavy chain CDR1 NOL13: GGATTCACCTTCAGAAGCTATGGC SEQ ID NO: 71 Nucleic acid sequence heavy chain CDR2 NOLI, NOL3, NOL4, NOL5, NOL6, NOL7, NOL8: ATATGGTATGATGGAAGTAATAAA SEQ ID NO: 72 Nucleic acid sequence heavy chain CDR2 NOL2: ATATGGTATGATGGAAGAAATAAA SEQ ID NO: 79 Nucleic acid sequence heavy chain CDR2 NOLI 1, NOL9, NOL10,NOL12: ATATCATATGATGGAAGTAATAAA SEQ ID NO: 83 Nucleic acid sequence heavy chain CDR2 NOL13: ATATCAAATGATGGAAGTAATAAA SEQ ID NO: 84 Nucleic acid sequence heavy chain CDR3 NOLI, NOL3, NOL4, NOL5, NOL6, NOL7: GCGAGAGGACTCTATGGGCGATCCTACTTTGACTAC SEQ ID NO: 85 Nucleic acid sequence heavy chain CDR3 NOL2: GCGAGAGGTCTCTTTGGGCGATCCTACTTTGACTAC SEQ ID NO: 91 Nucleic acid sequence heavy chain CDR3 NOL8: GCGAGAGGACTCTTTGGGCGATCCTACTTTGACTAC SEQ ID NO: 92 Nucleic acid sequence heavy chain CDR3 NOL9, NOL12, NOL13: GCGAAAGGGTTCAAGATACAGCTATGGTTTGCTGCTTTTGATATC SEQ ID NO: 93 Nucleic acid sequence heavy chain CDR3 NOL10: GTGAAAGGGTTCAAGATACAGCTATGGTTTGCTGCTTTTGATATC SEQ ID NO: 94 Nucleic acid sequence heavy chain CDR3 NOLI 1: GTGAAAGGGTTAAAGATACAGCTATGGTTTGCTGCTTTTGATATC SEQ ID NO: 97 Nucleic acid sequence light chain CDR1 NOL 1, NOL2, NOL3, NOL4, NOL5, NOL6, NOL8, NOLI 0: AACATAGGAAGTAAAAGT SEQ ID NO: 103 Nucleic acid sequence light chain CDR1 NOL7: AACATAGGAAGTAAGAGT SEQ ID NO: 105 Nucleic acid sequence light chain CDR1 NOLI 1, NOL9, NOL12: AACATTGGAAGTAAAAGT SEQ ID NO: 109 Nucleic acid sequence light chain CDR1 NOLI 3: AACATTGGTAGTAAAAGT SEQ ID NO: 110 Nucleic acid sequence variable light chain CDR3 NOLI, NOL3, NOL4, NOL8: CAGGTGTGGGATAGTGGTGATGGTCATCCGAGGGTG SEQ ID NO: 111 Nucleic acid sequence variable light chain CDR3 NOL2: CAGGTGTGGGATAGTGGTGATGGTCATCCTAGGGTG SEQ ID NO: 114 Nucleic acid sequence variable light chain CDR3 NOL5: CAGGTGTGGGATAGTGGTGATGGTAATCCGAGAGTG SEQ ID NO: 115 Nucleic acid sequence variable light chain CDR3 NOL6 : CAGGTGTGGGATAGTGGTGATGGTCATCCAAGGGTG SEQ ID NO: 116 Nucleic acid sequence variable light chain CDR3 NOL7: CAGGTGTGGGATAGTGGTGATGGTAATCCGAGGGTG SEQ ID NO: 118 Nucleic acid sequence variable light chain CDR3 NOL11, NOL9, NOL10, NOL12, NOL13: CAGGTGTGGGATAGTAGTAGTGATCCTGTGGTA SEQ ID NO: 123 Nucleic acid sequence variable heavy chain NOLI, NOL5: CAGGTACAGCTGCAGCAGTCAGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGACTCTATGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 124 Nucleic acid sequence variable heavy chain NOL2: CAGGTACAGCTGCAGCAGTCAGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACTTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATGGTATGATGGAAGAAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGTCTCTTTGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 125 Nucleic acid sequence variable heavy chain NOL3, NOL7: CAGGTACAGCTGCAGCAGTCAGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGTTGG AGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGACTCTATGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 126 Nucleic acid sequence variable heavy chain NOL4: CAGGTACAGCTGCAGCAGTCAGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATAAACAGTCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGACTCTATGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 128 Nucleic acid sequence variable heavy chain NOL6: CAGGTACAGCTGCAGCAGTCAGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AATGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGACGATTCACTATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGACTCTATGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCCCCTCA SEQ ID NO: 130 Nucleic acid sequence variable heavy chain NOL8: CAGGTACAGCTGCAGCAGTCAGAGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGCTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAGAGGACTCTTTGGGCGATCCTACTT TGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 131 Nucleic acid sequence variable heavy chain NOL9: GAGGTGCAGCTGGTGGAGACTGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAAAGGGTTCAAGATACAGCTATGGTT TGCTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCT TCA SEQ ID NO: 132 Nucleic acid sequence variable heavy chain NOL10: GAGGTGCAACTGGTGGAGACTGGGGGAGGCGTGGTCCGGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCATCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATATTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGATAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGTGAAAGGGTTCAAGATACAGCTATGGTT TGCTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCT TCA SEQ ID NO: 133 Nucleic acid sequence variable heavy chain NOLI 1: GAGGTGCAGCTGGTGGAGACTGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATG CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGTGAAAGGGTTAAAGATACAGCTATGGTT TGCTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACTGTCTCT TCA SEQ ID NO: 134 Nucleic acid sequence variable heavy chain NOLI 2: GAGGTGCAGCTGGAGGAGACTGGGGGAGGCGTGGTCCAGCCTGG GAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCGGT AGCTATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTC GAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTAT GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCC AAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGAC ACGGCTGTGTATTACTGTGCGAAAGGGTTCAAGATACAGCTATGG TTTGCTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCT CTTCA SEQ ID NO: 135 Nucleic acid sequence variable heavy chain NOL 13: GAGGTGCAGCTGGTGGAGACTGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGAA GCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCAAATGATGGAAGTAATAAATACTATG CGGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCA AGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACA CGGCTGTGTATTACTGTGCGAAAGGGTTCAAGATACAGCTATGGTT TGCTGCTTTTGATATCTGGGGCCAAGGGGCAATGGTCACCGTCTCT 5 TCA SEQ ID NO: 136 Nucleic acid sequence variable light chain NOLI: TCCTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGCCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTCTCTGGCTCCAACTCTGGCAACACGGCCACCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTCATCCGAGGGTGTTCGGCGGAGGGACCAAGT TGACCGTCCTA ID NO: 137 Nucleic acid sequence variable light chain NOL2 : TCCTATGAGCTGACTCAGCCACCCTCGATGTCAGTGGCCCCTGGAC AGACGGCCAGGATTGCCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTTTGATGACACCGACCGGCCCTCGGGGATCCCTGACCGGT TCTCTGGCTCCAACTCTGGCAACACGGCCACCCTGACCATCAGCAG GGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGA TAGTGGTGATGGTCATCCTAGGGTGTTCGGCGGAGGGACCAAGTT GACCGTCCTA SEQ ID NO: 138 Nucleic acid sequence variable light chain NOL3: TCCTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGCCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTGTCTGGCTCCAACTCTGGCAACACGGCCGCCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTCATCCGAGGGTGTTCGGCGGAGGGACCAAGT TGACCGTCCTA SEQ ID NO: 139 Nucleic acid sequence variable light chain NOL4: TCCTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGTCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTCTCTGGCTCCAACTCTGGCAACACGGCCACCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTCATCCGAGGGTGTTCGGCGGAGGGACCAAGT TGACCGTCCTA SEQ ID NO: 140 Nucleic acid sequence variable light chain NOL5: TCCTATGAGCTGATTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGCCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACGCCGACCGGCCCTCGGGGATCACTGACCGA TTCTCTGGCTCCAACTCTGGCAACACTGCCACCCTGACCATCAGCA GGGTCGAGATCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTAATCCGAGAGTGTTCGGCGGAGGTACCAAGT TGACCGTCCTA SEQ ID NO: 141 Nucleic acid sequence variable light chain NOL6: TCCTATAAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGCCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTCTCTGGCTCTAACTCTGGCAACACGGCCACCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTCATCCAAGGGTGTTCGGGGGAGGGTCCAAGT TGTCCGTCCTA SEQ ID NO: 142 Nucleic acid sequence variable light chain NOL7: TCCTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTGCCTGTGAGGGAGACAACATAGGAAGTAAG AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTCTCTGGCTCCAACTCTGGCAACACGGCCACCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTAATCCGAGGGTGTTCGGCGGAGGGACCAAGT TGACCGTCCTA SEQ ID NO: 143 Nucleic acid sequence variable light chain NOL8: TCCTATGAGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGAC AGACGGCCAAGATTACCTGTGAGGGAGACAACATAGGAAGTAAA AGTGTACACTGGTACCAGCAGAGGCCAGGCCAGGCCCCTGTGTTG GTCGTCTATGATGACACCGACCGGCCCTCGGGGATCCCTGACCGA TTCTCTGGCTCCAACTCTGGCAACACGGCCACCCTGACCATCAGCA GGGTCGAGGTCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTGGTGATGGTCATCCGAGGGTGTTCGGCGGAGGGACCAAGT TGACCGTCCTA SEQ ID NO: 144 Nucleic acid sequence variable light chain NOL9: CAGTCTGTGTTGACGCAGCCGCCCTCGGTGTCAGTGGCCCCAGGA CAGACGGCCAGGATTACCTGTGGGGGAAACAACATTGGAAGTAAA AGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTG GTCGTCTATGATGATAGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAGCA GGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTAGTAGTGATCCTGTGGTATTCGGCGGAGGGACCAAGCTGA CCGTCCTA SEQ ID NO: 145 Nucleic acid sequence variable light chain NOL10: CAGTCTGTGTTGACGCAGCCGCCCTCGGTGTCAGTGGCCCCAGGA CAGACGGCCAGGATTACCTGTGGGGGAAACAACATTGGAAGTAAA AGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTG GTCGTCTATGATGATAGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCCACTCTGACCATCGGCA GGGTCGAATCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTAGTAGTGATCCTGTGGTATTCGGCGGAGGGACCAAGCTGA CCGTCCTA SEQ ID NO : 146 Nucleic acid sequence variable light chain NOL 11: CAGTCTGTGTTGACGCAGCCGCCCTCGGTGTCAGTGGCCCCAGGA CAGACGGCCCGGATTACCTGTGGGGGAAACAACATTGGAAGTAAA AGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTG GTCGTCTATGATGATAGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAGCA GGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTAGTAGTGATCCTGTGGTATTCGGCGGAGGGACCAAGCTGA CCGTCCTA SEQ ID NO: 147 Nucleic acid sequence variable light chain NOLI 2: CAGTCTGTGTTGACGCAGCCGCCCTCGATATCAGTGGCCCCAGGA CAGACGGCCAGGATTACCTGTGGGGGAAACAACATTGGAAGTAAA AGTGTGCACTGGTACCATCAGAAGCCAGGCCAGGCCCCTGTGCTG GTCGTCTATGATGATAGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAGCA ATAGTAGTAGTGATCCTGTGGTATTCGGCGGAGGGACCAAGTTGA CCGTCCTA SEQ ID NO: 148 Nucleic acid sequence variable light chain NOL13: CAGTCTGTGTTGACGCAGCCGCCCTCGGTGTCAGTGGCCCCAGGA CAGACGGCCAGGATTACCTGTGGGGGAAACAACATTGGTAGTAAA AGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTG GTCGTCTATGATGATAGCGACCGGCCCTCAGGGATCCCTGAGCGA TTCTCTGGCTCCAACACTGGGAACACGGCCACTCTGACCATCAGCA GGGTCGAAGCTGGGGATGAGGCCGACTATTACTGTCAGGTGTGGG ATAGTAGTAGTGATCCTGTGGTATTCGGCGGAGGGACCAAGCTGA CCGTCCTA SEQ ID NO: 149 Nucleic acid sequence constant heavy chain: GCCAGCACCACCGCTCCTAAGGTGTTCCAGCTGGCCTCTCACTCTG CCGGCACAAGCGATTCTACAGTGGCCCTGGGATGTCTGGTGTCCA GCTACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCT GACATCTGGCGTGCACACATTTCCAAGCGTGCGGCAGTCTAGCGG CCTGTACTCTCTGAGCAGCATGGTCACAGTGCCTGCCAGCAGCCTG AAGTCCCAGACCTACATCTGCAACGTGGCCCATCCAGCCTCCAGC ACCAAGGTGGACAAGCGGATCGTGATCAAAGAGCCCTGCTGCTGC CCTAAGTGCCCCGACTCTAAGTTCCTGGGCAGACCCAGCGTGTTCA TCTTCCCACCTAATCCTAAGGACACCCTGATGATCAGCAGAACCCC TGAAGTGACCTGCGTGGTGGTGGACGTGTCCCAAGAGAACCCCGA CGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGCCCACACCGC CACAACAAAGGCCAAAGAGAAGCAGGACGCCAGCACCTACAGAG TGGTGTCCGTGCTGCCCATTCAGCAACCAGGATTGGCGGAGAGGC AAAGAATTCAAGTGCAAAGTGAACAATCGGGCCCTGCCTGCTCCT GTGGAACGGACAATCACAAAAGCCAAGGGCGAGCTGCAGGACCC CAAGGTGTACATTCTGGCCCCTCACCGGGAAGAAGTGACCAAGAA TACCGTGTCCGTGACCTGCCTCGTGAAGGACTTCTACCCTCCTGAC ATCAACGTGGAATGGCAGAGCAACGAGGAACCCGAGCCAGAAGT GAAGTACAGCACCACACCAGCTCAGCTGGATGGCGACGGCTCCTA CTTCCTGTACAGCAAGCTGACCGTGGAAACCGACAGATGGGAGCA GGGCGAGAGCTTCACCTGTGTGGTTATGCACGAGGCCATCCGGCA CACCTACCGGCAGAAGTCCATCACCAACTTTCCCGGCAAG SEQ ID NO: 150 Nucleic acid sequence constant light chain: GGCCCCACCAGCACACCTTCCGTGTCTCTGTTTCCTCCAAGCAGCG AGGAACTGAGCGCCAACAAGGCCACAGTCGTGTGCCTGATCAGCG ACTTTAGCCCCTCTGGCCTGGAAGTGATCTGGAAAGTGAACGACG CCGTGACCACCGACGGCGTGCAGACAACAAGAAGCAGCAAGCAG AGCAACGGGAAGTACGCCGCCAGCAGCTACCTGACAAGAACAAG CGCCCAGTGGAAGTCCTACAGCAGCGTGTCCTGCCAAGTGAAGCA CCAGGGCAAGACCGTGGAAAAGAAGCTGAGCCCTAGCGAGTGCC ec SEQ ID NO: 151 Amino acid sequence CH1 constant region of heavy chain: ASTTAPKVFQLASHSAGTSDSTVALGCLVSSYFPEPVTVSWNSGALTS GVHTFPSVRQSSGLYSLSSMVTVPASSLKSQTYICNVAHPASSTKVDK RI SEQ ID NO: 152 Nucleic acid sequence CH1 constant region of heavy chain: GCCAGCACCACCGCTCCTAAGGTGTTCCAGCTGGCCTCTCACTCTGC CGGCACAAGCGATTCTACAGTGGCCCTGGGATGTCTGGTGTCCAGC TACTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGAC ATCTGGCGTGCACACATTTCCAAGCGTGCGGCAGTCTAGCGGCCTG TACTCTCTGAGCAGCATGGTCACAGTGCCTGCCAGCAGCCTGAAGT CCCAGACCTACATCTGCAACGTGGCCCATCCAGCCTCCAGCACCAA GGTGGACAAGCGGATC SEQ ID NO: 153 Fab fragment heavy chain NOLI 1 amino acid sequence: EVQLVETGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW VAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY YCVKGLKIQLWFAAFDIWGQGTMVTVSSASTTAPKVFQLASHSAGTS DSTVALGCLVSSYFPEPVTVSWNSGALTSGVHTFPSVRQSSGLYSLSSM VTVPASSLKSQTYICNVAHPASSTKVDKRIVIKEPC SEQ ID NO: 154 Fab fragment heavy chain NOLI 1 nucleic acid sequence: GAGGTGCAGCTGGTGGAGACTGGGGGAGGCGTGGTCCAGCCTGGG AGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAG CTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAG TGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAG ACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAA CACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCT GTGTATTACTGTGTGAAAGGGTTAAAGATACAGCTATGGTTTGCTGC TTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTATCTTCAGCC AGCACCACCGCTCCTAAGGTGTTCCAGCTGGCCTCTCACTCTGCCGG CACAAGCGATTCTACAGTGGCCCTGGGATGTCTGGTGTCCAGCTAC TTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACATC TGGCGTGCACACATTTCCAAGCGTGCGGCAGTCTAGCGGCCTGTAC TCTCTGAGCAGCATGGTCACAGTGCCTGCCAGCAGCCTGAAGTCCC AGACCTACATCTGCAACGTGGCCCATCCAGCCTCCAGCACCAAGGT GGACAAGCGGATCGTGATCAAAGAGCCCTGC SEQ ID NO: 155 Light chain CDR 2 (NOL1-NOL13): DDXi, wherein Xi can be S, T or A, preferably Xi is S. SEQ ID NO: 156 Light chain CDR 2 (NOLI 1,NOL9,NOL10,NOL12,NOL13): DDS. ID NO: 156 is the most preferred light chain CDR2. SEQ ID NO: 157 Light chain CDR 2 (NOLI, 2, 3, 4, 6, 7, 8): DDT. SEQ ID NO: 158 Light chain CDR 2 (NOL5): DDA. SEQ ID NO: 159: Nucleic acid sequence variable region light chain CDR2 (NOLI, NOL2, NOL3, NOL4, NOL6, NOL7, NOL8): GATGACACC SEQ ID NO: 163 Nucleic acid sequence variable region light chain CDR2 NOL5: GATGACGCC SEQ ID NO: 167 Nucleic acid sequence variable region light chain CDR2 (NOLI 1, NOL9, NOL10, NOL12, NOL13): GATGATAGC. The isolated antibody or antibody derivative may comprise at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 1, 155, 156, 157, 158,2,3,4,5,6,7,8,9,10, 11,12,13,14,15, 16, 17, 18, 19, 20 and 21. Preferably, the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155,156,2,6,7, 10, 11, 15 and 16. In another preferred embodiment, the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155, 2, 6, 10 and 15. In another preferred embodiment, the isolated antibody or antibody derivative comprises all complementary determining region (CDR) amino acid sequences as depicted in any one of SEQ ID NOs: 1, 155,2,6,10 and 15. In a particularly preferred embodiment, the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 1, 156, 2, 7, 11 and 16. In a highly preferred embodiment, the isolated antibody or antibody derivative comprises all complementary determining region (CDR) amino acid sequences as depicted in any one ofSEQ ID NOs: 1, 156, 2, 7, 11 and 16. In another preferred embodiment of the invention, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155, 156, 157 and 158, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3, 4 and 5, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6, 7, 8 and 9, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 10, 11, 12, 13 and 14, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as in any one of SEQ ID N0s:15, 16, 17, 18, 19, 20 and 21. More preferably, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155, 156 and 157, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3 and 4, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6 and 7, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 10, 11 and 13, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID N0s:15, 16, 17, 19 and 20. Also more preferably, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 156 and 157, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2 and 4, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in SEQ ID NO: 7, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 11 and 13, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 16, 17 and 20. Evenmore preferably, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence as depicted in SEQ ID NO: 155, a light chain CDR3 amino acid sequence as depicted in SEQ ID NO: 2, a heavy chain CDR1 amino acid sequence as depicted in SEQ ID NO: 6, a heavy chain CDK2 amino acid sequence as depicted in SEQ ID NO: 10 and a heavy chain CDR3 amino acid sequence as depicted in SEQ ID N0:15. Most preferably, the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence as depicted in SEQ ID NO: 156, a light chain CDR3 amino acid sequence as depicted in SEQ ID NO: 2, a heavy chain CDR1 amino acid sequence as depicted in SEQ ID NO: 6, a heavy chain CDR2 amino acid sequence as depicted in SEQ ID NO: 11 and a heavy chain CDR3 amino acid sequence as depicted in SEQ ID N0:16. The isolated antibody or antibody derivative optionally comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23, 24, 25, 26,27,28,29, 30, 31, 32,33,35,36 or 37, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39, 40, 41, 42,43, 45, 49, 50, 51, 52 or 53. Preferably, the isolated antibody or antibody derivative comprises a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23 or 24, and/or comprises a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39 or 40. More preferably, the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in any one of SEQ ID NOs: 22, 23 or 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in any one of SEQ ID NOs: 38, 39 or 40. Even more preferably, the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in SEQ ID NO: 23 or 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in SEQ ID NO: 39 or 40. Most preferably, the isolated antibody or antibody derivative comprises a light chain variable region amino acid sequence as depicted in SEQ ID NO: 24, and/or comprises a heavy chain variable region amino acid sequence as depicted in SEQ ID NO: 40. In a highly preferred embodiment, the isolated antibody or antibody derivative according to the invention comprises a light chain amino acid sequence as depicted in SEQ ID NO: 54, and/or comprises a heavy chain amino acid sequence as depicted in SEQ ID NO: 55 or 153, more preferably as in SEQ ID NO: 55. The invention also provides isolated nucleic acid molecules, vectors and host cells comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to the invention. Optionally, the at least one nucleic acid sequence is selected from the group consisting of the nucleic acid sequences as depicted in any one ofSEQ ID NO: 58, 59,69,70,71,72, 79,83, 84, 85, 91, 92, 93, 94, 97, 103, 105, 109,110,111, 114, 115, 116, 118,123, 124,125,126, 128, 130-150, 152, 154, 159, 163 and 167. Preferably, the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO: 58, 79,94,105,118,133,146, 149,150,152,154 and 167. More preferably, the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO: 58, 79, 94, 105,118 and 167. In another preferred embodiment, the isolated nucleic acid molecule, vector and/or host cell of the invention comprises at least a nucleic acid sequence selected from a group consisting of the nucleic acid sequences as depicted in any one SEQ ID NO:133,146,149,150 and 152. In another preferred embodiment, all ofSEQ ID NO: 133, 146, 149 and 150 are used to produce the antibody or antibody derivative of the present invention on one or more vectors or in on or more host cells. Most preferably, the nucleic acid sequences SEQ ID NOs: 149 or 152 and 150 encode the constant regions of the heavy and light chain of the antibody or antibody derivative according to the invention, respectively. In the case of antibody derivatives, truncated constant regions and, thus, truncated forms of SEQ ID NOs: 149, 150 and/or 152 may be used. NOLI 1 is the most preferred antibody. As explained above, a method of manufacturing of the isolated antibody or antibody derivative according to the invention is provided, comprising expressing the isolated antibody or antibody derivative in a host cell. Preferably, the isolated antibody or antibody derivative is expressed in a host cell according to the invention, such as a eukaryotic cell. The isolated nucleic acid molecules, vectors and host cells comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to the invention as described above are particularly useful in the method of manufacturing of the isolated antibody or antibody derivative according to the invention and may be used for expressing the antibody or antibody derivatives. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Titration ELISA of affinity-maturated antibodies from wildtype NOLI on eqIL-5. Figure 2: Titration ELISA of affimty-maturated antibodies from wildtype NOL2 oneqIL-5. Figure 3: Inhibition effect of antibodies on the binding of eqIL-5 to its receptor. The antibodies were preincubated with the antigen and then added to Expi293F cells which present the eqIL-5 receptor on their surface. Binding of the antigen to the cells was detected in the flow cytometer by fluorescent labelling via the His-tag and a fluorescently labelled antibody. To obtain the relative binding, the measured fluorescence signal (median) was related to the signal when only antigen and cells, but no inhibiting antibody, were present. Figure 4: HPLC-SEC data of NOLI 1 after storage at 4 °C, 21 °C and 37 °C for 2 or 4 weeks, respectively. Figure 5: ELISA results of NOLI 1 after storage at 4 °C, 21 °C or 37 °C for 2 weeks or 4 weeks. Figure 6: Reducing (left) and non-reducing (right) SDS-PAGE of samples of NOLI 1 after storage at 4 °C, 21 °C and 37 °C. Figure 7: SDS-Page stained with Coomassie (left) and Western Blot stained with Streptavidin-HRP (right) of NOLI 1, produced in Hi5 insect cells and Expi293F™ mammalian cells. Aglycosylated samples containing the N297A mutation and glycosylated samples were analyzed. Figure 8: Timeline of the target animal study D10POC-A. Three horses received the antibody (42.62 |^g antibody/kg body weight) and the other three horses received PBS on day 0 and 14 of the study. Blood samples were taken on day -1 (as background control), day 2, day 7, day 14 before the second injection, day 16, day 21, day 28 and day 120. These samples were collected and used for analysis (ADA assay). Figure 9: Anti-Dmg antibody (ADA) assay comparison of the target animal study D10POC-A: Comparison of antibody and control group for all six horses up to 120 days after injection. Horse semm was titrated on coated NOL11 in eqFab format and detected with anti-horse IgG (Fc-)HRP antibody. The OD450nm-620nm values from day -1 were subtracted from the values of all other time points to analyze signal change over time. The average AOD450nm-620nm signal of the three horses of the antibody group was compared to the average signal of the three horses of the control group at a serum dilution of 1:100. Figure 10: Timeline of the target animal studies D10POC-B and D10POC-C. Injection of NOLI 1 took place at day 0, 30 and 60. Figure 11: Anti-Dmg antibody (ADA) assay of target animal study D10POC-B: Evaluation ofAnti-Drug-Antibody formation in the horses treated with NOL11. The two-center study D10POC-B took place in Eastern and Northern Germany. The study in Eastern Germany is still on going, therefore, only the data from the Northern Germany study is included. The antibody group consists of two horses treated with NOL11. The control group consists of two horses that were not treated with the antibody. Injection of NOLI 1 took place at day 0, 30 and 60. Figure 12: Skin scores of insect bite hypersensitivity (IBH) affected horses (study D10POC-C, day -7-91). The severity ofIBH was determined for each horse before the trial (mild, moderate, or high grade IBH). Data show the skin scores between 0 (no abnormality) and 3 (severe itch/skin lesions) evaluated during the 91-day study period (starting 7 days prior the first injection). The horses were scored in the first five days after each injection and then every second day. (a) Skin scores of the untreated control horses (No.329) classified as low to moderately affected by IBH. (b) Skin score of the moderately affected horse No. 344 treated with the anti-eqIL-5 antibody NOLI 1. (c) Skin score of the severely affected horse No.306 treated with NOLI 1. The antibody (0.05 mg/kg) was administered subcutaneously at three time points (day 0, day 30 and day 60). Figure 13: The graph shows the linear regression analysis of the skin score (Fig.12) of the three study horses over the observation period of 91 days. The severity of insect bite hypersensitivity (IBH) was determined for each horse before the trial (low, moderate, or high grade IBH). The antibody (0.05 mg/kg) was administered subcutaneously at three time points (day 0, day 30 and day 60). Figure 14: Skin scores of insect bite hypersensitivity (IBH) affected horses (study D10POC-C, day 91 -135). The severity ofIBH was determined for each horse before the trial (low, moderate, or high grade IBH). Data show the skin scores between 0 (no abnormality) and 3 (severe itch/skin lesions) evaluated during day 91 and day 135 after the last injection of NOL11. The horses were scored every second day. The antibody (0.05 mg/kg) was administered subcutaneously at three time points (day 0, day 30 and day 60). (a) Skin scores of the untreated control horse (No.329) classified as low to moderately affected by IBH. (b) Skin score of the moderately affected horse No.344 treated with NOLI 1. (c) Skin score of the severely affected horse No.306 treated with NOLI 1. Figure 15: The graph shows the linear regression analysis of the skin score of the three study horses evaluated between day 91 and day 135 after the last injection of NOLI 1. The antibody (0.05 mg/kg) was administered subcutaneously at three time points (day 0, day 30 and day 60). The severity of IBH was determined for each horse before the trial (mild, moderate, or high grade IBH). Figure 16: Scratching behavior in horses treated with NOLI 1 (0.05 mg/kg, n = 2) or in an untreated control (n =1) (study D10POC-C). The severity of IBH was determined for each horse before the trial (mild, moderate, or high grade IBH). The equine pruritus visual analogue scale (EPVAS) was used to determine the scratching behavior. The scale is graduated from 0 (normal horse; no itch) to 10 (extremely severe itching). The antibody (0.05 mg/kg) was administered subcutaneously at three time points (April, May and June; four weeks between each in injection). The observation started 21 days after the last injection of NOLI 1 (theoretical half-life of equine antibodies). Figure 17: Overview CDR1, CDR2 and CDR3 of the variable light chains of NOL1-13. Figure 18: Overview CDR1, CDR2 and CDR3 of the variable heavy chains of NOL1-13. EXAMPLES 1. Monoclonal antibodies against the summer eczema of horses Monoclonal antibodies against eqIL-5 were generated via antibody phage display. Afterwards, the format of the binders was changed from single chain variable fragment (scFv) to full length immunoglobulin G (IgG). For the constant domains, an equine sequence was used. Out of the seven equine IgG subclasses, the subtype IgG6 was chosen due to its lowest effector function. The antibodies were produced in mammalian cells (Expi293F™) and validated by Enzyme-linked Immunosorbent Assay (ELISA) and a self-established cellular inhibition assay using flow cytometry. Two final candidates that show strong inhibition in the cellular assay, were chosen for further consideration. Those two candidates were affinity-maturated in vitro. The mutants were tested and compared in ELISA and inhibition assay. 1.1 Materials and Methods 1.1.1 Production ofrecombinant antibodies Antibody production was performed in mammalian Expi293F™ cells (Thermo Fischer). These are cultivated at 37 °C, 110 rpm and 5% C02 in F17 medium (8 mM L-glutamine and 0,1% Pluronic-F68 added). A transient expression was used for the production. The transfection of expression plasmids was performed with linear polyethylene imine (PEI MAX, 40 kDa) (Polysciences Europe GmbH). Heavy and light chain of the antibody were cloned into different expression plasmids which were co-transfected in a ratio of 1:1. The production scale was adjusted depending on the required protein amount and ranged between 7.5 mL to 125 mL. Independent of the scale, between 1.5 - 2.0 x 106 cells/mL were used. They were transfected with plasmid DNA and PEI per milliliter of cell culture volume. DNA and PEI were first diluted separately and then mixed in F 17 medium (10% of the culture volume) and incubated at room temperature for around 30 min. Then, the DNA-PEI mix was added to the cells. The cells were cultivated under the conditions mentioned above for 7 days in total, including a feeding step after 48 h with new medium. The cells were harvested via centrifugation. Subsequently, the supernatant was sterile filtered for purification. Antibodies were purified from the culture supernatant using affinity chromatography via binding to protein A or protein G. Depending on the scale, different columns were installed using the Akta or Profinia system or a 24-well filter plate and compressed air was utilized. 7.7.2 Titration ELISA The binding of the antibodies to eqIL-5 was detected by ELISA. For this purpose, eqIL-5 was immobilized in a 96-well plate, the corresponding antibodies were added and then detected with an antibody directed against the equine Fc part. As a negative control for non-specific binding, the antibody was also added to BSA. In detail, 100 ng antigen per well or 1%BSA solution was immobilized in a 96-well plate. Nonspecific binding sites were then blocked with milk powder before the antibody was added. MPBST (PBS with 0.05% Tween and 2% milk powder) was used for blocking as well as for dilution of all antibodies. After each step, each well was washed three times (ELISA Washer 405™ LS, BioTek™) to remove unbound components. A horseradish peroxidase (HRP)-coupled antibody from Sigma-Aldrich was used as detection antibody (SAB3700145, dilution 1:5000). Binding was detected by adding TMB substrate and subsequent HRP-catalyzed blue staining of the corresponding well. The reaction was stopped with sulfuric acid and the optical density was measured at 450 mn and 620 nm with an ELISA reader. To obtain binding curves of the antibodies, a titration from 316 nM to 0.0036 nM with the dilution factor 1/^/10 was performed and then the optical density (OD450nm- OD620nm) was plotted against the antibody concentration. The measurements were performed in triplicates. A fit with the "logistic5" function using the software OriginPro was used to determine EC50 values. 1.1.3 Cellular inhibition assay A cellular iiAibition assay was developed in order to test whether the selected antibodies not only bind the antigen but can also inhibit the binding of the interleukin to its receptor. The corresponding equine IL-5 receptor was presented on the surface of the cells and the binding of the antigen was detected via flow cytometry. Equine eosinophil cells would have been suitable for this purpose, as they express the receptor for eqIL-5. However, as there is no equine eosinophil cell line and isolation is difficult to reproduce and not possible in a large scale, Expi293F™ suspension cells were used instead and transiently transfected with the receptor DNA. The receptor was not secreted due to a fusion to the transmembrane domain, but presented on the cell surface. The eqIL-5 receptor consists of two subunits, the IL5RA and CSF2RB, which were co-transfected. In addition, the cells were co-transfected with eGFP to differentiate transfected from non-transfected cells. Transfection was performed as described for antibody production. Cells were then cultured for 48 h at 37 °C, 110 rpm and 5% C02. After 48 h, the cells were used for the inhibition assay. The measurements were performed in the flow cytometer. The antigen was fluorescently labelled via an anti His-tag antibody and a secondary detection antibody that is fluorescently labelled. Thus, binding of the antigen could be visualized by a fluorescent cell. A decrease in fluorescence indicated inhibition of binding. A background measurement was performed using neither antigen nor inhibiting antibody, as well as a reference measurement with antigen but no inhibiting antibody. In detail, all dilutions and washing steps during cell preparation were carried out in FACS buffer (1 x PBS + 2% FCS + 5 mM EDTA). First, the antibodies to be tested were diluted to a concentration of 2 |J.M and titrated in a 96-well plate with a factor of 1 / ^10. The antigen was added to the antibodies in the same volume and at a concentration of 20 nM, so that the antibody had a final maximum concentration of 1 pM and the antigen a final concentration of 10 nM. The antibody and antigen were preincubated for 45 minutes at room temperature. Meanwhile, cells transfected with receptor DNA and eGFP were pelleted and adjusted to a cell density of5xl06 cells/mL in FACS buffer.5 x 105 cells/well were placed in a 96-deepwell plate and washed with FACS buffer. The cell pellet was resuspended in the antibody-antigen mix and incubated on ice for about 1 hour. After another washing step, the cell pellet was taken up in primary antibody (Penta His antibody: Qiagen, 3460, 1:50 dilution) and again incubated on ice for about 1 h. This was followed by another washing step and resuspension in the secondary detection antibody (goat anti mouse Fc-APC: dianova, 115-136-071, 1:50 dilution). After further incubation and two washing steps, the cells were resuspended in FACS buffer and measured with the flow cytometer (MACS Quant). First, the cells were gated. Living single cells were selected and of these cells only the transfected cells (GFP positive) were considered. The APC signal was measured from these cells and the median was used for the evaluation. The background signal, generated by non-specific binding of the primary and secondary antibodies to the cells, was subtracted from all measured values. The reference indicates the APC value, which is generated by the binding of the antigen (eqIL-5-His) to the cells without inhibition. In the evaluation, this value corresponds to 100% binding. In the case of an inhibitory antibody, this prevents the antigen from binding to the cell, which reduces the APC signal accordingly. These APC values are related to the 100% of the APC value of the reference control and thus give the relative binding of the antigen to the cell in percent. The more inhibitory the antibody, the lower the relative binding. A titration of the antibodies was performed to determine the relative binding as a function of antibody concentration. The IC50 value, which characterizes the antibody concentration at 50% inhibition, provides a suitable benchmark. 1.2 Results 1.2.1 Titration ELISA Figure 1 and Figure 2 show the results of the titration ELISA. Two different antibodies against eqIL-5 (NOLI und NOL9), which behave differently in ELISA, were chosen for in vitro affinity maturation. The two following graphs show the affinity-maturated antibodies in comparison to their respective wildtype. All chosen antibodies show specific binding to eqIL-5 and no binding to BSA. Both, the antibodies based on NOLI (Figure 1) and based on NOL9 (Figure 2) demonstrated good binding activities. The affinity maturation was successful in finding antibodies with even improved binding activity compared to the wildtype. The following Table 1 shows the corresponding EC50 values, which may all be considered as suitable for the purpose of the invention and the therapeutic application of the antibody of the invention. They were determined with the OriginPro software using the "logistic 5" fit. Table 1: EC50 values of the antibodies against eqIL-5 determined by titration ELISA
Figure imgf000057_0001
The values are all in a similar, nanomolar range. The lowest value is 0.44 nM, the highest 8.99 nM. For the NOLI antibody the affinity maturation improved the £€50 value by factor 2.2 and for the NOL9 antibody the EC50 value was improved by factor 1.4. 7.2.2 Cellular inhibition assay Since the titration ELISA only informs about the binding of the antibodies to the antigen, an additional assay to test the inhibition was performed. The goal was to analyze the antibodies' ability to inhibit the binding of the antigen to its receptor on a cell surface. Figure 3 presents the inhibition effect of the antibodies against eqIL-5. The y-axis describes the relative binding of the interleukin to its receptor. The antibodies partially inhibit the binding completely, depending on their concentration. The curves of the different antibodies are very similar. The antibodies derived after affinity maturation of both wildtypes are equally effective in inhibition, despite slightly different affinities in the titration ELISA (Figures 1 and 2). This is possibly explained by the fact that the antibodies recognize different epitopes. The curves were determined with the "logistic 5" fit using the OriginPro software. Table 2 shows the IC5o values which are the antibody concentrations needed to reduce the binding to 50%. A reduction to 50% binding is achieved at concentrations between 3.0 nM and 9.4 nM. For the NOLI antibody the affinity maturation improved the IC50 value by factor 2.0 and for the NOL9 antibody the IC50 value was improved by factor 1.9.
Table 2: IC50 values of the antibodies against eqIL-5 determined by cellular inhibition assay
Figure imgf000059_0001
2. Characterization of a chimeric-equine anti-ILS antibody Further experiments were performed with NOLI 1, a preferred antibody. 2.1 Materials and Methods 2.1.1 Stability tests To determine the long-term stability of the antibody NOLI 1, different aliquots were stored at 4°C, 21°C and 37°C. After two weeks of storage, new aliquots were added. After two additional weeks, the samples were analyzed by HPLC-SEC, ELISA and SDS-PAGE in comparison with freshly thawed material to obtain data for two and four weeks of storage at the given temperatures. 2.1.1.1 HPLC-SEC For HPLC-SEC, the antibodies were diluted to a concentration of 50 |J.g/mL in PBS- NaNs. They were analyzed on an Agilent AdvanceBio SEC. 27.7.2 ELISA To determine an effect on the binding to the antigen eqIL-5, after storage at 4 °C, 21 °C and 37 °C a Titration ELISA was performed as described previously. Briefly, eqIL-5 was coated onto a 96-well plate. After blocking and washing, the samples of NOLI 1 were titrated on it. Unbound antibodies were washed away, and an anti-equine antibody was added for detection. 2.1.1.3 SDS-PAGE The samples were also analyzed by SDS-PAGE under reducing and non-reducing conditions. Reducing samples were mixed with Laemmli buffer with P-mercapto- ethanol and heated for 5 min at 56 °C while non-reducing samples were mixed with Laemmli buffer without P-mercaptoethanol and heated at 56 °C for 10 min. The electrophoresis was performed at 150-200 V for approximately 1 hour. 2.1.2 Biotinylation for prove of aglycosylation Usually, antibodies contain an N-glycosylation site in the Fc-part. We introduced an aglycosylation mutation into our anti-IL5 antibodies to remove the glycosylation site. By replacing the asparagine (N) of the glycosylation site with an alanine (A), glycosylation is prevented in the Fc part of the antibody. The aglycosylation mutation is called N297A based on the fact that human antibodies are glycosylated at the position 297. To show, that the mutagenesis was successful, and no glycosylation was added to NOLI 1, a biotinylation experiment was performed. As a positive control, a glycosylated version of NOLI 1 was used. This experiment was performed with NOLI 1 recombinantly produced in Expi293F™ cells as described previously and Hi5 cells (described below). Firstly, the antibodies were prepared by changing the buffer via gel filtration to 100 mM NaAc/150 mM Nad, pH 5.5. Then they were incubated in sodium-periodate solution for oxidation of sialic acid, a common sugar component of protein polysaccharides. The buffer was again changed by gel filtration. As the next step Hydrize-PEG4-Biotin was added to the samples (final cone.5 mM) and incubated. Excess biotin was removed by gel filtration and the samples were used for Western Blot Analysis. 2.7.2.7 Western Blot Analysis In Western Blot analysis, the biotin was specifically detected. After a reducing SDS- PAGE was performed, the proteins were blotted onto a membrane. Unspecific binding sites were blocked and Streptavidin-HRP was used for chemiluminescence detection. 2.1.2.2 Production of recombinant antibodies in Hi5 cells The cells were cultivated at 27 °C and 120 rpm. A transient expression was used for the production. The transfection of expression plasmids was performed with linear polyethylene imine (PEI MAX, 40 kDa) (Polysciences Europe GmbH). Heavy and light chain of the antibody were cloned into different expression plasmids, which were co-transfected in a ratio of 1:1. The transfection was performed in a volume of 15 mL.4.0 x 106 cells/mL were used. They were transfected with plasmid DNA and PEI per milliliter of cell culture volume. DNA and PEI were directly added to the cells. The cells were diluted to a concentration of 1.0 x 106 cells/mL and cultivated under the conditions mentioned above for 4 days in total, including a feeding step after 48 h with new medium. The cells were harvested via centrifugation. Subsequently, the supernatant was sterile filtered for purification. Antibodies were purified from the culture supernatant using affinity chromatography via binding to protein A or protein G. Depending on the scale, different columns were installed using the Akta or Profinia system or a 24-well filter plate and compressed air was utilized. 2.2 Results 2.2.7 Stability tests analyze the stability of NOL 11, HPLC-SEC, SDS-PAGE and ELISA experiments were performed with samples stored at 4 °C, 21 °C and 37 °C. 2.2.7.7 HPLC-SEC In the HPLC-SEC, for each sample only one peak is visible. Despite a small difference in height, all peaks look comparable and have their maximum at a retention time of approximately 12.5 minutes. No additional peaks at a shorter or longer elution time are visible. This leads to the conclusion, that no aggregates or fragments are present in the samples after 4 weeks of storage at the respective temperatures (Figure 4). 227.2 ELISA In the ELISA, all samples show comparable curves to the fresh antibody. Only the two samples, which were stored at 21 °C have a slightly higher maximum value at a concentration of 100 nM. The ability of NOLI 1 to bind eqIL-5 does not change over a period of 4 weeks with storage at either 4 °C, 21 °C or 37 °C (Figure 5). 2.2.1.3 SDS-PAGE All samples were analyzed by SDS-PAGE under non-reducing conditions, the samples after 4 weeks of storage were additionally analyzed by reducing SDS-PAGE. Under non-reducing conditions, all samples show a band between 150 and 250 kDa which consists of two different bands. This fits to the size of a full IgG. Additionally, a small band is visible above the main band. This is probably aggregated antibody. The aggregates are also present in the fresh sample. When the antibody is stored at 37 °C, slightly more aggregated antibody can be detected. Under reducing conditions in all analyzed samples, two bands are visible at 25 kDa and 50 kDa. The disulfide bonds between the heavy and light chain of the antibody are broken and we see one band for the heavy chain (50 kDa) and one band for the light chain (25 kDa). The sample, which was stored at 37 °C, shows two slight additional bands and around 100 kDa and 150 kDa. No big differences are visible between the samples, leading to the conclusion, that the antibody is stable under all tested conditions (Figure 6). 2.2.2 Biotinylation for prove of aglycosylation Our antibodies were modified by the N297A mutation, called aglycosylation, to have no N-glycosylation in the Fc-part. A biotinylation experiment was performed to show that no sugars are bound to the antibody NOL11, the antibody used for two in vivo studies. For this experiment, NOL11 from two different expression systems was used. In addition to Expi297F™ cells, the antibody was produced in Hi5 insect cells. As a positive control, the glycosylated form of NOLI 1 without the mutation was used. The sialic acid of potential protein polysaccharides was oxidized and coupled to Hydrazide-PEG4-Biotin. In a reducing SDS-PAGE and Western Blot, the biotin was detected by Streptavidin-HRP. The results are presented in Figure 7. On the SDS-PAGE stained with Coomassie, all four antibodies showed two bands. They fit to the size of the heavy chain (50 kDa) and light chain (25 kDa). On the western blot, only two of those bands are visible. The heavy chains of the glycosylated proteins were stained with Streptavidin-HRP. This proves, that the N297A mutation successfully led to aglycosylation of the antibody (Figure 7). 3. Target animal studies with NOLI 1 Due to promising in vitro results in the inhibition assay as well as producibility and stability data, the anti-eqIL-5 antibody NOL11 tested in vivo in horses (healthy and IBH affected). NOLI 1 has been tested for safety and tolerability in two target-animal studies to date (D10POC-A and D10POC-B). In the second study, the efficacy of the antibody was additionally tested in a small number of animals suffering from IBH (D10POC-C). An overview of each study with relevant information on the animals involved is provided in Table 3. Two healthy control animals that received placebo or were untreated are not included in Table 3. Table 3: Designation of the target-animal studies A, B and C with NOL11 BW: Body weight; IBH: insect bite hypersensitivity.
Figure imgf000064_0001
Figure imgf000065_0001
3.1 D10POC-A, single-center safety study (Northern Germany) / orientation study A For the in vivo orientation study D 1 OPOC-A, six horses from the own herd at the WDT serum factory in Memsen, Germany, were used. The primary aim of the study was the safety evaluation of the antibody NOL11. Therefore, four healthy horses and two horses affected by IBH were used. The horses were divided in an antibody and a control group each containing two healthy and one sick horse. The control group received 1 x PBS instead of the antibody. An overview of the test horses is given in Table 3. 3.1.1 Medical history of the horses affected by summer eczema The horse 306 arrived at the WDT with already diagnosed summer eczema. For horse No.329, no information is available about the start of the disease at the arrival. The summer eczema of horse No.306 is classified as high-grade, while the horse No.329 has low-grade (mild to moderate) summer eczema. Both horses were normally stabled during the main flight period of the mosquitos to avoid exposure to Culicoides. Both summer eczema horses were treated with the repellent Wellcare™ three times a week during the mosquito season (April/May- October). In addition, the affected skin areas (mane crest, tail mmp and belly seam) were rubbed with Ballistol animal to soothe the skin and ward off the insects. When the mare No.306 with the high-grade summer eczema was kept in an open stable two years ago, this led to a severe aggravation of the symptoms despite the above-mentioned treatment. Highly increased itching led to a complete loss of hair in the area of the tail rump and the belly seam. Also, inflammatory swellings and chafed, sore skin occurred in the area of the tail mmp, mane crest and the entire underside of the belly. Most of the mane hair was rubbed away and the rest broke off. As a conclusion, the horse was moved back into the stable with daily exercise. The gelding No.329 with low-grade summer eczema was always stabled with daily turnout. Its symptoms were most noticeable on the tail rump and belly seam. Only some of the mane hair was broken off, the skin in this area was mostly unharmed. For ethic reasons this horse was used as control animal and not the mare No.306. 3.1.2 Design of the orientation study Dl OPOC-A In this in vivo orientation study six horses were examined. Three horses belonged to the antibody group (horse No.306, horse No.365 and horse No.376) receiving the NOLI 1 antibody solved in 1 x PBS buffer (antibody concentration 1.86 mg/mL) and the other three horses (horse No.329, horse No.312 and horse No.383) belonged to the control group that received Ix PBS instead of antibody. The applied amount of antibody (42.62 |J.g/kg) was calculated as a 1000 x molar access to an average eqIL-5 level (100 pg/mL, self-determined by serum ELISA). The antibody or 1 x PBS was subcutaneously injected twice, on day 0 and day 14 of the study. The study started in April before the start of the IBH season. For the analysis, blood samples were taken on day -1 (as background control), day 2, day 7, day 14 before the second injection, day 16, day 21, day 28 and day 120. In addition, a score sheet monitoring the horses' health and behavior was filled in on days 0, 1, 2, 3, 5, 7, 9, 11,13,14,15,16, 17,19, 21,23,25,27,29, 31,33,35,37,39, 41 and 120. During the study, the horses were kept in an open stable with other horses. In addition, the horses with IBH had not been treated with Wellcare™ Repellent and Ballistol Animal-Oil since the last mosquito season and during the study. Figure 8 visualizes the timeline of the study. 3.1.3 Evaluation of safety 3.1.3.1 Score sheet The score sheets were filled in by the attending veterinarians on the days mentioned above. Following parameters were evaluated: 1: known side effects of Cytopoint (anaphylaxis, urticaria, facial oedema, diarrhea, ataxia, central nervous disturbance, convulsions), 2: increase of body temperature after injection, 3: swelling at the injection site, 4: pain at the injection site, 5: abscess formation at the injection site, 6: appetite, 7: habitus, 8: gait, 9: fur, 10: body condition score and 11: symptom expression of summer eczema. During the three days after the injections all parameters were monitored, after that only the parameters 6-11 were considered. Each parameter was scored between 0 (no abnormality) and 3 (severe indications). Table 4 provides an overview of the total score for each horse on every observation day. Table 4: Results of the score sheets
Figure imgf000068_0001
SE: Summer eczema symptoms; U: Urticaria The only abnormality was the development of urticaria of horse No.312 shortly after injection of 1 x PBS. Apart from the appearance shortly after the injection, however, no rational connection could be established. As the animal was also in the pasture at the time of the appearance, insect bites cannot be excluded as the causative agent here. Further, with the score 1 this was just a minor effect which only lasted for two days. In the applied doses and volumes, the antibody NOLI 1 showed no side effects. The summer eczema symptoms did not exceed a score of 1 (minor itching with occasional mbbing, at a maximum broken hairs, hardly disturbance of the general condition, normal feed intake and herd behavior) for neither of the two summer eczema horses up to day 41. Horse No.306 had a score 1 on six days (days 13 to day 19) while horse No.329 had a score 1 on four days (day 14 to day 16 and day 23). The results indicate that both summer eczema horses had symptoms in a comparable range. This is a positive outcome considering that horse No.306 showed much stronger symptoms in the previous years compared to horse No.329, which indicates a good efficacy of the antibody. On day 120, horse No.306 had a total score of 3 and horse No.329 a total score of 1. This time point is outside the time period, in which the antibody influences the symptoms. Here, horse No.306 is stronger affected by summer eczema as it was the case in the previous years. Table 5 shows a comparison of the summer eczema scores and symptoms of former years with the symptoms of the study year (after treatment).
Table 5: Comparison of severity of summer eczema symptoms of horse No.306 and No.312
Figure imgf000070_0001
Figure imgf000071_0001
Taken together, there was no observation of adverse side effects of the NOL11 antibody. Therefore, a high safety and tolerability is assumed for this product. Concerning the efficacy, one has to mention that in the study year in general the symptom development of horses with summer eczema seemed to be reduced. Furthermore, the number of the used horses with summer eczema in this study does not allow statistically significant evaluation of the efficacy. Nevertheless, taken the fact that horse No.306 had more severe symptoms over the last years compared to horse No.329, one can state a tendency of efficacy of NOLI 1 treatment since the symptoms of the two horses were similarly low during the study period. 3.1.3.2 Anti-drug-antibody (ADA) assay In the target animal studies, a chimeric antibody with human-derived variable domains and equine constant domains was injected. The human domains could be recognized by the immune system of the horse as foreign. To test whether antibodies against NOL11 were generated and therefore causing an unwanted immune reaction in the treated horse, an ADA assay with the horse semm was performed. The ADA assay was performed via ELISA. Here, 100 ng of the NOLI 1 in Fab format (The Fab format was chosen to avoid unspecific binding of the detection antibody to the Fc-part in the ELISA assay. Binding of serum antibodies to the Fc-part of the antibody is not expected since the Fc-part is a constant region that is part of the equine immune system) were coated per well in a high-binding 96-well plate at 4 °C overnight. As controls also an alternative eqFab antibody and 2% BSA were used. Then, wells were blocked with 2% BSA in PBS-T for 1 h at room temperature and washed three times using an ELISA washer (BioTek™ 405™ LS). In the next step, the horse serum was titrated with a starting dilution of 1/10 and a dilution factor of 1/VTH and incubated for 1 h at room temperature. After washing again, the detection antibody goat IgG anti horse IgG (Fc-)HRP (SAB3700145, Lot RI36383, Sigma- Aldrich, dilution 1:20000 in 2% BSA) was applied for 1 h at room temperature. Wells were washed again and TMB substrate was added to start the coulometric reaction and 1 M sulfuric acid was used to stop the reaction. The absorbance (OD450nm-620nm) was measured with an ELISA reader (Epoch, Biotek). All measurements were performed in duplicates. For the analysis, the signals of the serum sample day -1 were subtracted from the signal of all other serum samples (AOD450nm-620nm) so that only the change but not the absolute value is evaluated. For a comparison between the antibody group and the control group, the average AOD450nm-620nm signals were compared at a serum dilution of 1:100 (Figure 9). Here, no significant difference between the two groups was detected. For the antibody group, the AOD450nm-620nm signal from day 14 onwards is increased up to approximately 0.06 while the signal of the control group fluctuates closely around 0. This is still in the expected fluctuation range and does not indicate the development of ADAs. If the horse generated ADAs it would happen after the second dose with some delay in time. Up to 120 days no significant increase of the signal of the antibody group compared to the control group was detected. The slight signal of the antibody group from day 14 on is still in a range that lies within the expected fluctuations. Furthermore, no ADAs are expected before administration of the second dose, indicating that a signal increase as at day 14 has no relevance. 3.2 D I OPOC-B; two-center safety study (Northern and Eastern Germany) 3.2.1 Study design Five horses received NOLI 1 solved in Ix PBS buffer at a dosage of 0.05 mg/kg bodyweight. One horse served as untreated control. All horses suffered from IBH according to their medical history. Horses were housed in groups in open stables. The antibody was administered subcutaneously at three time points (day 0, day 30 and day 60). The horses received their first injection mid of April, followed by the second injection in May and the last and third one in June (30 days between each in injection). A score sheet monitoring the horses' general health and behavior was filled in by the supervising veterinarian regularly. In center 1 (Northern Germany), the horses were scored in the first five days after each injection and then every second day. In center 2 (Eastern Germany), the horses were scored in the first three days after injection following by scoring once a week. The safety was evaluated according to the parameters described in chapter 3.1.3 for the study D10POC-A. The last score sheet for the safety evaluation was completed on day 90 and therefore 30 days after the last injection. Blood samples were taken on day 0, 2, 3, 30,32,33,60,62, 63,90,91, 120 and 121. The ADA assay was performed via ELISA as described for the orientation study A (D10POC-A, chapter 3.1.3.2). Horse No.306 was a part of the studies D10POC-A and D10POC-B and C, therefore receiving the antibody a second year in row. As the likelihood of ADA formation may increase with the number of injections, horse No.306 was carefully screened for adverse immune reactions. The study in Eastern Germany is still on-going, therefore, only the samples from the horses in Northern Germany were analyzed so far. 3.2.2 Results 3.2.2.1 Scoring The parameters assessed (anaphylaxis, urticaria, facial oedema, diarrhea, ataxia, central nervous disturbance, convulsions, increase of body temperature after injection, swelling at the injection site, pain at the injection site, abscess formation at the injection site, appetite, habitus, gait, fur, body condition score) were scored in 5 horses over the entire study period. Only one of these horses showed a three-day swelling of the injection site 5 days after the 3 injection (score 2 on day 5 after injection and score 1 in the following two days). The other horses showed no side effects. 3.2.2.2 Determination of Anti-Drug-Antibodies (ADAs) An ADA assay was performed (as already described in chapter 3.1.3.2) to test whether antibodies against NOLI 1 were generated and therefore causing an unwanted immune reaction in the treated horse. For comparison between the NOL11 group and the control group, the average AOD450nm-620nm signals (the OD450nm-620nm of day are subtracted from the OD450nm-620nm of the respective sample to obtain AOD450nm-620nm as a value for the deviation from sample before application of NOLI 1) were compared at a serum dilution of 1:100 (Figure 11). Here, no significant difference between the two groups was detected. For the antibody group, the AOD450nm-620nm signal of day 90 and day 91 is increased up to approximately 0.09 and 0.10 while the signal of the control group fluctuates closely around 0. This is still within the expected fluctuation range and does not indicate the development ofADAs. On days 3 and 32 for example, the signal of the antibody group decreases in the same range to approximately -0.09, showing that this is a normal fluctuation range. Horse No.306 showed no immune response to of NOLI 1 the second year in a row with a total of five injections (two in D10POC-A, three in D10POC-B). This indicates that the antibody is safe for a repeated treatment of horses. 3.3 D10POC-C; single-center efficacy study (Northern Germany) 3.3.1 Study design During the study D10POC-B, two horses treated with NOLI 1 as well as an untreated control horse were intensively monitored for the expression of symptoms (referred to as study D10POC-C). The assessment was carried out by the veterinary who has cared for the horses for several years and knows their medical history very well. Before the study started, the severity of IBH of the study horses was assessed, based on clinical expression in previous years. One horses (Alma, no.306) was classified as having severe sweet itch (high grade IBH), one horse (Wescana, No.344) was classified as moderate, and the untreated control horse (Tamino, No.329) was classified as having a mild to moderate expression of IBH symptoms. The study started in April, shortly after the first symptoms appeared. All horses were kept in the same group in an open stable without any special protection against midges. A skin score sheet monitoring the horses' scratching behavior and skin condition was filled in by the veterinary starting one week before the first inj ection and then regularly as described in chapter 3.2.2.1. The last score sheet for the efficacy evaluation was completed on day 135 and therefore 75 days after the last injection. The evaluation was divided into two observation periods: Day -7 to Day 91 (i.e. up to 31 days after the last injection) and Day 91 to Day 135 (period after the expected duration of action of NOL11 based on the theoretical half-life of equine antibodies (Lewis et al., Molecular Immunology, 2008, 45, 818-827). Anti-human IL-5 antibodies such as Resilizumab or Mepolizumab show a long-term pharmacodynamic effect after the treatment with the antibody in asthma patients that is far beyond the semm half-life of the antibodies (Ghassemian et al. , Allergy, Asthma and Clinical Immunology 2021, 17, 1-7; Smith e? a/., Clinical Pharmacokinetics 2011, 50, 215-227; Wang et al., CPT: Pharmacometrics and Systems Pharmacology 2017, 6, 249-257). Therefore, the period after the last injection was of high interest. Horses were scored between 0 (no signs of pruritus/skin lesions) and 3 (severe signs of pmritus/skin lesions). As control, a healthy horse without IBH was scored to due signs of IBH in parallel. This horse was scored with "0" at every time point (data not shown). In addition, 21 days after the last injection (theoretical half-life of equine antibodies (Lewis et al., 2008)), the Equine Pruritus Visual Analog Scale (EPVAS) was recorded to obtain a trend for the pharmacodynamic effect of NOLI 1. This score has values from 0 (normal horse) to 10 (extremely severe itching) (Craig et al.. Journal of the American Veterinary Medical Association 2023, 261, 75-85). 3.3.2 Results (Skin scores and EP VAS) Figure 12 shows the skin scores over the first 91-day observation period. The first score data were collected 7 days prior to the first injection of NOLI 1 (day -7 to -1). Figure 12a shows the score of the untreated control horse. At the start of the trial in April (day -7 to day -5) the score was mostly 0 in this mild to moderate affected horse (Fig.12a, average score 0.1). Horse No.306 (Fig.12c), classified as having very severe IBH (high grade), showed already mild symptoms before the first injection in April (average score 1). Horse No.344 (Fig.12b), classified as moderately affected by IBH, showed mild symptoms before the first NOLI 1 injection (average score 0.4). Comparing the average scores of the untreated control horse and the two NOLI 1 treated horses between day 0 (day of the first NOLI 1 injection at the end of April) and day 91 (31 days after the third and final dose ofNOL 11), the treated horses had a lower average score than the untreated control horse. Horse No.344 (Fig.12b) had an average score of 0.1 between day 0 and day 91, whereas the average score of the control horse (horse No.329, Fig.12a) was 1.0. The average score for severely affected horse No.306 was 0.7 and therefore below the score of the control horse. Fig.13 shows the regression line of the scores over the 91-day observation period. The increase in the untreated horse is the most prominent, while the line of the moderately affected horse hardly increases. The increase in the severely affected horse treated with NOLI 1 is lower than for the untreated control animal. Looking at the period from day 91 to day 135 (75 days after the last dose ofNOLll) between the end of July and the beginning of September, there is a worsening of symptoms in all horses (Fig.14). However, the average score of the treated animals is still below the average score of the control horse. The average score results were score 2 for the mild to moderate affected control horse (Fig.14a) and score 1.8 (Fig.14c) and score 0.4 (Fig.14b), respectively, for the severely and moderately affected horses treated with NOLI 1. Interestingly, the severely affected horse No.306 treated with NOL11 showed a greater increase in symptoms than the control horse, which can be explained by the waning effect of the antibody. However, the treated horses were still significantly less symptomatic than known from previous years, suggesting that inhibition of IL-5 at the start of the season has a sustained effect throughout the season (see also case reports in chapter 3.4). Prolonged or intense scratching damages the skin, resulting in inflammatory processes, which in turn promotes pruritus. This creates a vicious circle of pruritus and mechanical irritation (itch-scratch cycle). If this cycle of itching and scratching is interrupted at the very beginning, it is very likely that the maintenance and worsening of the disease will be delayed. During May, June, July and August, the months when symptoms typically begin to manifest, the average score was 1.0 in the severely affected horse No.306, which was below the level of the untreated control horse, known as a mild to moderate sweet itch sufferer (average sore 1.4). Horse No.344 treated with NOLI 1 showed almost no to only very mild symptoms throughout the study. Only 33 days after the last dose of NOLI 1, this horse began to show prolonged symptoms (average score May-August 0.3). Fig.15 shows the regression line of the skin scores from day 91 on until the last score at day 140. Both regression lines of the treated horses show a comparable increased slope after the treatment period and a steep ascent of the line compared to the control horse. Fig.16 shows the development of scratching behavior 21 days after the last injection of NOLI 1 (dose 3). Scratching behavior was scored from 0 to 10 using the EPVAS score. In the untreated control horse as well as in the NOL 11-treated highly affected IBH horse, pruritus increases constantly 21 days after the last dose. However, the NOL 11-treated horse with severe IBH (No.306) did not show more severe pruritus than the untreated horse, known to have mild to moderate disease, even 81 days after the last antibody injection. Horse No.344 started to scratch 39 days after the last NOLI 1 injection but showed only minor scratching behavior until the end of the study. 3.4 Case reports At the end of studies D10POC-A and C, a case report was written for the two horses treated with NOLI 1 by the attending veterinary. 3.4.1 Case I.' severe IBH horse Case study horse 1 ("Alma", No.306), a 17-year-old warmblood mare, suffers from IBH for more than 10 years. The horse exhibits a high degree ofpmritus without protective measures (stabling for most of the day, reduced grazing, daily application of skin care oil and regular application of the repellent permethrin). Symptoms usually start in April and last until November, depending on climatic conditions (warm weather and high humidity are ideal conditions for midges). The horse developed numerous skin lesions up to open sores on various parts of the body due to the intense scratching behavior. Particularly affected are the mane crest, head, tail mmp and abdominal seam. Itchy lesions can also be found on the udder, inner thighs, elbow crease and chest. Affected skin areas on the tail mmp show derb-elastic skin changes ("elephant skin") due to the continuous scratching behavior. The horse had to be treated several times a week with permethrin (Wellcare™ Emulsion 10.5 mg/ml) and with skin oil daily. The horse suffers from such severe pruritus that regular application of glucocorticoid ointments and systemic dexamethasone injections are required during the sweet itch season. During the studies, the horse was kept in an open stable with daily access to pasture. The horse was not treated with repellents, skin oil or glucocorticoids. In study D10POC-A, the horse received two injections of NOL11 (0.04 mg/kg bodyweight) two weeks apart (April and May) before the onset of first symptoms. At the beginning of May, the first, but only mild, symptoms appeared. The horse showed slight chafing around the mane crest and sheared hair in the area of the tail rump. There were no skin lesions or open wounds. At the beginning of July, approximately 80 days after the last NOL11 injection, the expression of the symptoms became somewhat more marked, and the horse showed more distinct rubs in the mane and tail area as well as some hairless areas on the belly suture. However, there were still no visible skin lesions or wounds. The horse did not require any further protective or therapeutic measures during the test season (no repellents, stabling, reduced grazing, regular stabling, skin oil or glucocorticoids) and still showed a significantly lower expression of summer eczema compared to previous years. In study D10POC-C, the horse received three injections of NOLI 1 (0.05 mg/kg bodyweight; 30 days interval in April, May and June) after the onset of symptoms. This was done to see if it is possible to down-regulate an already existing symptomatic condition. The horse showed only minor symptoms until mid-June. At this time point, the horse showed more pronounced rubbing and was treated with skin oil for two consecutive days. After the third NOL11 injection, the symptoms weakened again. In the middle of July, there was again a worsening of the symptoms. The horse showed moderate itching reflected in mbbed off long hair. Itchy skin swellings were visible at the abdominal seam. However, at no time were there clear skin lesions or open wounds. Apart from application of skin oil on two days, no further measures were necessary during the NOL 11-therapy. Overall, the IBH was much less pronounced than in the years without NOL 11 treatment. 3.4.2 Case 2: moderate IBH horse Case study horse 2 ("Wescana", No.344), a 19-year-old warmblood mare, suffers from IBH for more than 10 years. The horse is moderately affected by IBH without protective measures (stabling for most of the day, reduced grazing, daily application of skin care oil and regular application of the repellent permethrin). The horse shows increasing chafing over the season, especially on the mane crest and tail rump. The treatment consists of weekly application of permethrin (Wellcare™ Emulsion 10.5 mg/ml) and care of the affected areas with skin oil. No glucocorticoid treatment is needed. Horse 2 participated only in studies D10POC-B and C. Analogous to horse 1 (chapter 3.4.1), horse 2 received three injections ofNOLll (0.05 mg/kg bodyweight; 30 days interval in April, May and June) after the onset of symptoms. The horse was kept in an open stable with daily access to pasture. The horse was not treated with repellents or skin oil throughout the study. During the complete treatment period (3 injections), horse 2 showed no signs ofIBH except for a breakout a few days before the third injection at the end of June. Approximately 40 days after the last injection, itching symptoms could be observed again, but only minor rubbing on the mane and tail rump resulting in broken hair. In contrast, the untreated control horse ("Tamino", No.329) showed more pronounced symptoms during studies C and B, than in previous years. It is known that the expression of symptoms can vary from year to year, depending on the climatic conditions and insect flight in the respective year. In summary, NOL11 significantly reduced the severity of symptoms in the two case study horses suffering from IBH for more 10 years.

Claims

CLAIMS 1. An isolated antibody or antibody derivative that specifically binds to equine interleukine-5 (IL-5) comprising at least one member selected from a group consisting of equine constant light chain, equine constant heavy chain and fragments thereof. 2. The isolated antibody or antibody derivative according to claim 1, wherein the isolated antibody or antibody derivative is a monoclonal isolated antibody or antibody derivative. 3. The isolated antibody or antibody derivative according to claim 1 or 2, wherein the isolated antibody or antibody derivative is aglycosylated. 4. The isolated antibody or antibody derivative according to any one of claims 1 to 3, wherein the isolated antibody or antibody derivative comprises at least one complementary determining region (CDR) amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQIDNOs: 1,155,156,157,158,
2,
3,
4,5,6,7,8,9,10,11,12,13,14,15, 16,17,18,19,20 and 21.
5. The isolated antibody or antibody derivative according to any one of claims 1 to 4, wherein the isolated antibody or antibody derivative comprises a light chain CDR1 amino acid sequence as depicted in SEQ ID NO: 1, a light chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 155, 156, 157 and 158, a light chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 2, 3, 4 and 5, a heavy chain CDR1 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one of SEQ ID NOs: 6, 7, 8 and 9, a heavy chain CDR2 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID NOs: 10,11, 12, 13 and 14, and a heavy chain CDR3 amino acid sequence selected from a group consisting of the amino acid sequences as depicted in any one ofSEQ ID N0s:15,16,17,18,19,20and21.
6. The isolated antibody or antibody derivative according to any one of claims 1 to 5 comprising a light chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs:22,23,24,25, 26,27,28,29,30,31, 32,33,35,36 or 37, and/or comprising a heavy chain variable region comprising an amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to an amino acid sequence as depicted in any one of SEQ ID NOs:38,39,40,41, 42,43,45,49,50, 51, 52 or 53.
7. The isolated antibody or antibody derivative according to any one of claims 1 to 6 comprising a light chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 54, and/or comprising a heavy chain amino acid sequence being identical to or having at least 90%, 95%, 98% or 99% sequence homology to the amino acid sequence as depicted in SEQ ID NO: 55 or 153.
8. The isolated antibody or antibody derivative according to any one of claims 1 to 7, wherein the isolated antibody or antibody derivative is an isolated IgG antibody or antibody derivative, preferably, the isolated antibody or antibody derivative is an isolated IgG6 antibody or antibody derivative.
9. The isolated antibody or antibody derivative according to any one of claims 1 to 8, wherein the isolated antibody or antibody derivative has a neutralizing ability towards equine IL-5.
10. An isolated nucleic acid molecule comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to any one of claims 1 to 9.
11. A vector comprising at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to any one of claims 1 to 9, preferably the vector is an expression vector.
12. A host cell comprising the vector according to claim 11 or at least one nucleic acid sequence encoding at least partially the isolated antibody or antibody derivative according to any one of claims 1 to 9, preferably, the host cell is a eukaryotic host cell.
13. A pharmaceutical composition comprising the isolated antibody or antibody derivative according to any one of claims 1 to 9 and a pharmaceutically acceptable excipient.
14. The isolated antibody or antibody derivative according to any one of claims 1 to 9 or the pharmaceutical composition according to claim 13 for use as a medicament.
15. The isolated antibody or antibody derivative according to any one of claims 1 to 9 or the pharmaceutical composition according to claim 13 for use in the treatment of an allergic condition of a horse.
16. The isolated antibody or antibody derivative for use according to claim 15 or the pharmaceutical composition for use according to claim 15, wherein the allergic condition is selected from a group consisting of equine allergic skin disease, equine allergic respiratory disease and inflammatory disease.
17. The isolated antibody or antibody derivative for use according to claim 16 or the pharmaceutical composition for use according to claim 16, wherein the equine allergic skin disease is selected from a group consisting of insect-bite hypersensitivity, atopic dermatitis, food hypersensitivity, allergic and irritant contact dermatitis and urticaria, preferably, the equine allergic skin disease is insect-bite hypersensitivity or atopic dermatitis, most preferably, the equine allergic skin disease is insect-bite hypersensitivity.
18. The isolated antibody or antibody derivative for use according to claim 16 or the pharmaceutical composition for use according to claim 16, wherein the equine allergic respiratory disease is equine asthma.
19. An in vitro method for the diagnosis of an allergic condition in a horse comprising contacting the isolated antibody or antibody derivative according to any one of claims 1 to 9 with a biological sample.
20. A method of manufacturing the isolated antibody or antibody derivative according to any one of claims 1 to 9, comprising expressing the isolated antibody or antibody derivative in a host cell, preferably in a host cell according to claim 12.
PCT/EP2023/078860 2022-10-17 2023-10-17 Antibodies and antibody derivatives against equine il-5 WO2024083847A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP22202013 2022-10-17
EP22202013.3 2022-10-17

Publications (1)

Publication Number Publication Date
WO2024083847A1 true WO2024083847A1 (en) 2024-04-25

Family

ID=83898466

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/078860 WO2024083847A1 (en) 2022-10-17 2023-10-17 Antibodies and antibody derivatives against equine il-5

Country Status (1)

Country Link
WO (1) WO2024083847A1 (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019228990A1 (en) * 2018-05-28 2019-12-05 Evax Ag Treatment of urticaria

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019228990A1 (en) * 2018-05-28 2019-12-05 Evax Ag Treatment of urticaria

Non-Patent Citations (29)

* Cited by examiner, † Cited by third party
Title
"Gene Expression Systems - Using Nature for the Art of Expression", 1998, ACADEMIC PRESS
"Recombinant Protein Expression in Mammalian Cells", vol. 1850, 2018, THOMAS REINARD
"Textbook on Cloning, Expression and Purification of Recombinant Proteins", 2022, SPRINGER
BEEKMAN ET AL., J VET INTERN MED, vol. 26, no. 1, 2012, pages 153 - 161
BOND ET AL., J VET INTERN MED, vol. 32, no. 6, 2018, pages 2088 - 2098
BOND ET AL., RMC VET RES, vol. 15, 2019, pages 397
CORDEAU ET AL., VET. IMMUNOL., vol. 97, 2004, pages 87 - 96
CRAIG ET AL., JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION, vol. 261, 2023, pages 75 - 85
DEWACHI ET AL., VET IMMUNOL IMMUNOPATHOL, vol. 109, no. 1-2, 2006, pages 31 - 6
FADOK, VET. CLIN. EQUINE, vol. 29, 2013, pages 541 - 550
FETTELSCHOSS-GABRIEL ANTONIA ET AL: "Molecular mechanisms and treatment modalities in equine Culicoides hypersensitivity", VETERINARY JOURNAL, BAILLIERE TINDALL, LONDON, GB, vol. 276, 17 August 2021 (2021-08-17), XP086786725, ISSN: 1090-0233, [retrieved on 20210817], DOI: 10.1016/J.TVJL.2021.105741 *
FETTELSCHOSS-GABRIEL ET AL., ALLERGY, vol. 74, no. 3, 2019, pages 572 - 582
FETTELSCHOSS-GABRIEL ET AL., THE VETERINARY JOURNAL, vol. 276, 2021, pages 105741
FETTELSCHOSS-GABRIEL ET AL., VETERINARY JOURNAL, vol. 276, 2021, pages 105741
GHASSEMIAN ET AL., ALLERGY, ASTHMA AND CLINICAL IMMUNOLOGY, vol. 17, 2021, pages 1 - 7
IVESTER ET AL., J VET INTERN MED, vol. 28, no. 3, 2014, pages 918 - 924
JANSSEN ET AL., FRONTIERS IN IMMUNOLOGY, vol. 13, 2022, pages 921077
LANGREDER NORA ET AL: "Development of an inhibiting antibody against equine interleukin 5 to treat insect bite hypersensitivity of horses", SCIENTIFIC REPORTS, vol. 13, no. 1, 10 March 2023 (2023-03-10), US, XP093117457, ISSN: 2045-2322, Retrieved from the Internet <URL:https://www.nature.com/articles/s41598-023-31173-y> DOI: 10.1038/s41598-023-31173-y *
LAVOIE ET AL., AM. J. RESPIR. CRIT. CARE MED., vol. 164, 2001, pages 1410 - 1413
LEWIS ET AL., MOLECULAR IMMUNOLOGY, vol. 45, 2008, pages 818 - 827
MITSON-SALAZARPRUSSIN, FRONTIERS IN MEDICINE, vol. 4, 2017, pages 165
NAGASE ET AL., ALLERGOLOGY INTERNATIONAL, vol. 69, 2020, pages 178 - 186
NEDOROST, DERMATOLOGIC CLINICS, vol. 38, no. 3, 2020, pages 301 - 308
RIIHIMAKI ET AL., CAN J VET RES, vol. 72, no. 5, 2008, pages 432
ROUFOSSE FLORENCE: "Targeting the Interleukin-5 Pathway for Treatment of Eosinophilic Conditions Other than Asthma", FRONTIERS IN MEDICINE, vol. 5, 6 April 2018 (2018-04-06), XP055780050, DOI: 10.3389/fmed.2018.00049 *
ROUFOSSE, FRONT. MED., vol. 5, pages 49
SIMOES, J, EQUINE VET. SCI., vol. 87, 2020, pages 102937
SMITH ET AL., CLINICAL PHARMACOKINETICS, vol. 50, 2011, pages 215 - 227
WANG ET AL., CPT.-PHARMACOMETRICS AND SYSTEMS PHARMACOLOGY, vol. 6, 2017, pages 249 - 257

Similar Documents

Publication Publication Date Title
AU2018203261B2 (en) Engineered anti-TGF-beta antibodies and antigen-binding fragments
US20200370051A1 (en) Mrgprx2/mrgprb2 expressing cell based assay to detect pseudo-allergic drug reactions and to identify blockers to prevent the adverse reactions
US10584180B2 (en) Anti-AGE antibodies for treating inflammation and auto-immune disorders
US10336831B2 (en) Use of anti-endoglin antibodies for treating ocular fibrosis
JP4188353B2 (en) Enhanced vaccine
BR112020017703A2 (en) INTERLEUKIN-31 MONOCLONAL ANTIBODIES FOR VETERINARY USE
WO2021031716A1 (en) Anti-pd-l1 single domain antibodies
DE69126607T2 (en) EXTRA CELLULAR PIECES OF HUMAN IGE IMMUNOGLOBULIN ANCHOR PEPTIDES AND SPECIFIC ANTIBODIES THEREFOR
JP2014529295A (en) Interleukin-31 monoclonal antibody
US9926375B2 (en) Anti-endoglin antibodies and uses thereof
US9474794B2 (en) Antibodies that bind to adrenomedullin and to adrenomedullin receptors and the uses thereof as drugs
DE3887031T2 (en) Form of administration for an antagonist of gamma interferon for controlling a GHK-associated autoimmune disease.
CN105408355A (en) Methods for increasing immunoglobulin A levels
BR112020017715A2 (en) PEPTIDE VACCINES AGAINST INTERLEUCIN-31
CN110678196B (en) anti-HEKR-K envelope antibody and application thereof
US20060292138A1 (en) Allergen vaccine proteins for the treatment and prevention of allergic diseases
WO2024083847A1 (en) Antibodies and antibody derivatives against equine il-5
US20220273793A1 (en) Horse il-31 induced pruritus model
JP2022172170A (en) Therapeutic anti-IgE antibodies and methods and compositions thereof
JP2022172170A5 (en)
DE69922873T2 (en) Specific binding proteins for the treatment of allergies in dogs
EP3568411B1 (en) Methods and compositions for treating hpa hyperactivity
US12134660B2 (en) Anti-age antibodies for treating inflammation and auto-immune disorders

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23793714

Country of ref document: EP

Kind code of ref document: A1