RU2807067C2 - Antibodies against cxcr2 and their use - Google Patents

Abstract

FIELD: biochemistry.

SUBSTANCE: invention is related to a human antibody molecule that binds in an immune-specific way to human CXCR2. A nucleic acid encoding said antibody; a vector or cell containing said nucleic acid; a pharmaceutical composition containing said antibody, are also disclosed. Method of treatment using said antibody; use of said antibody to produce a medicinal product; a method of blocking neutrophil chemotaxis using said antibody; a method of blocking CXCR2 signalling in response to CXCL1 and/or CXCL5 in cells expressing CXCR2 signalling, are disclosed.

EFFECT: invention makes it possible to effectively treat diseases such as neutrophilia of the respiratory tract or acute pneumonia.

25 cl, 12 ex, 19 tbl, 15 dwg

Description

Cross reference to related applications

This application claims priority to U.S. Provisional Patent Application No. 62/713,095, filed August 1, 2018, the contents of which are incorporated herein by reference in their entirety.

List of sequences

This application contains a sequence listing that was filed electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy created on July 30, 2019 is called 102085_005304_SL and is 279,927 bytes in size.

Field of technology to which the invention relates

The present application relates to human antibody molecules that bind immunospecifically to human CXCR2.

State of the art

The most common white blood cells in the blood are neutrophils. They are important effector cells of the innate immune system and play a major role in the elimination of extracellular pathogens. However, if neutrophil recruitment to tissue is not properly controlled, chronic neutrophil infiltration and activation can lead to the continuous release of inflammatory mediators and proteinases that cause distinct tissue damage.

Neutrophil migration and activation is mediated by the interaction of CXC chemokine receptor-1 (CXCR1) and CXC chemokine receptor-2 (CXCR2) on the neutrophil plasma membrane with the ELR+CXC chemokines (CXCL1, 2, 3, 5, 6, 7, and 8). CXCR2 acts as a high-affinity receptor for all ELR+CXC chemokines and plays a key role in the mobilization and recruitment of neutrophils and monocytes from the blood to tissues. And the chemokines CXCL6, 7 and 8 also interact with CXCR1, which modulates oxidative burst activity and protease release from neutrophils, which is critical for immunity to bacteria and fungi.

Increased sputum neutrophil counts have been associated with phenotypes associated with increased asthma severity, corticosteroid insensitivity, and chronic respiratory difficulty. Airway neutrophilia increases with exacerbation of asthma. Airway neutrophilia is also common to all clinical phenotypes of chronic obstructive pulmonary disease (COPD), including emphysema-predominant COPD, frequent exacerbation COPD, and high eosinophil COPD. The degree of airway neutrophilia also correlates with the severity of the disease and the rate of physiological decline. Neutrophil proteinases, especially neutrophil elastase, are involved in all pathological manifestations of COPD. Proteinases secreted by neutrophils are also associated with the development of emphysema, promote the destruction of the extracellular matrix, and are associated with mucus hypersecretion. These associations suggest that neutrophil infiltration into the airways may play a critical role in the pathophysiological processes underlying severe asthma and COPD.

Chemokines CXCL1, CXCL5 and CXCL8 bind to CXCR2 and are involved in neutrophil recruitment. CXCL1, CXCL5 and CXCL8 are increased in chronic airway inflammation and are increased in sputum or bronchial biopsies from subjects with severe neutrophilic asthma or COPD. Counteracting CXCR2 activation by chemokines offers a potential therapeutic strategy by reducing neutrophil recruitment to tissue and neutrophil-mediated pathologies associated with these inflammatory diseases.

However, chemokine receptors have proven to be difficult targets for selective antagonism. Despite difficulties in developing compounds with the required target specificity and antagonistic activity, several small molecule CXCR2 antagonists have been effective in animal models of inflammation. In human clinical trials, small molecule CXCR2 antagonists did not show broad efficacy in subjects with neutrophilic asthma or COPD, although in a study of ozone and lipopolysaccharide inhalation-induced neutrophilia in sputum, they showed marked efficacy in otherwise healthy subjects. There was only a small improvement in baseline lung function (FEV ₁ ) in COPD patients who continued to smoke compared with former smokers. To date, all published clinical trials have used small molecule CXCR2 antagonists. The best studied is danirixin (GSK 1325756), a reversible and selective CXCR2 antagonist (CXCL8 binding _IC50 = 12.5 nM) which has also been shown to block CD11b up-regulation in neutrophils (e.g., see Miller et al. , BMC Pharmacol Toxicol 2015, 16: 18). Danirixin failed to meet primary objectives for COPD in a phase IIb trial. Other CXCR2-selective molecules include SB-566933 (Lazaar et al., Br. J. Clin. Pharmacol. 2011, 72:282-293) and AZD5069, which is selective for CXCR2 (>150-fold less active at CXCR1 receptors and CCR2b) and does not affect CD11b expression induced by C5a, LTB4 or fMLP (Nicolls et al., J. Pharmacol. Exp. Ther. 2015, 353: 340-350). Molecules that inhibit both CXCR2 and CXCR1 include navarixin (SCH 527123, MK-7123; Todd et al., Pulm Pharmacol Ther. 2016 Dec, 41:34-39) and ladarixin (DF2156A; Hirose et al., J Genet Syndr Gene Ther. 2013, S3). These molecules are being studied for many indications, including COPD, asthma and other inflammatory lung diseases, cancer and more.

In some studies, the use of small molecule CXCR2 antagonists resulted in a marked, undesirable decrease in blood neutrophil levels (neutropenia), which may limit the tolerated dose of such agents. Neutropenia may result from the antagonist being less specific for CXCR2 and/or from the antagonist acting on all CXCR2-binding ligands. CXCL8 and related CXC chemokines, for example, play an important role in the mobilization of mature granulocytes into the peripheral blood, so strong antagonism of these ligands on CXCR2 may interfere with normal migration of neutrophils into the blood. Conversely, preferential antagonism of the downstream pathway involving calcium signaling following ligand binding to CXCR2 may counteract unwanted neutrophil migration into the lungs while maintaining the desired ability of neutrophils to mobilize into the blood.

The essence of the invention

Human antibody molecules are provided that bind immunospecifically to human CXCR2. These human antibody molecules are more selective CXCR2 antagonists than currently described small molecule CXCR2 antagonists, more potent antagonists of CXCL1 and CXCL5-induced CXCR2 activation than currently described CXCR2 antagonist antibodies, and antagonize neutrophil recruitment to tissues without severely depleting circulating numbers. neutrophils. Human antibody molecules contain heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 167 and light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 168 or heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 226 and CDR1, CDR2 and Light chain CDR3 of SEQ ID NO: 227 and inhibit activation of human CXCR2 by human CXCL1 or human CXCL5. In some embodiments, the human antibody molecules are capable of inhibiting CXCR2 activation by CXCL1 or CXCL5 in a subject without causing severe persistent neutropenia.

Pharmaceutical compositions containing human antibody molecules are also provided.

Also provided are nucleic acid molecules encoding human antibody molecules, vectors containing nucleic acid molecules, and cells transformed to express human antibody molecules.

Also provided are methods for preventing or treating neutrophilia in peripheral tissues of subjects such as airway neutrophilia. Methods of reducing the level of monocytes in peripheral tissues of subjects are also provided. Methods for reducing eosinophilia in peripheral tissues of subjects are also provided.

Also provided are methods of reducing acute airway inflammation, methods of preventing or reducing chronic airway inflammation such as in bronchiectasis, methods of reducing tumor burden, methods of stopping or slowing the growth of cancer, methods of reducing chronic pain, methods of preventing or reducing neuroinflammation such as in diffuse sclerosis, ways to reduce inflammation in the liver, ways to reduce inflammation in the pancreas, or ways to reduce the symptoms of type I diabetes. The methods include administering to subjects a therapeutically or prophylactically effective amount of any of the specified human antibody molecules or any of the disclosed pharmaceutical compositions for the treatment or prevention of the specified diseases in the subjects.

Also provided are these human antibody molecules or pharmaceutical compositions for use in the prevention or treatment of airway neutrophilia or acute pneumonia. The use of human antibody molecules or pharmaceutical compositions in the manufacture of medicinal products for the prevention or treatment of airway neutrophilia or acute pneumonia is also contemplated.

Brief description of the figures

The spirit of the invention as well as the following detailed description will become clearer when read in conjunction with the accompanying drawings. For the purpose of illustrating the disclosed human antibody molecules, methods, and uses, the drawings show typical embodiments of human antibody molecules, methods, and uses; however, the molecules, methods and uses of human antibodies are not limited to the specific embodiments given.

In fig. 1A and 1B show the results of a representative dose-response study of inhibition of CXCL8-induced CXCR2 activation by representative disclosed anti-CXCR2 antibodies as measured in Tango™ cells.

In fig. 2 is a dot plot showing the binding activity of exemplary described anti-CXCR2 antibodies to native human CXCR2 expressed in human neutrophils and native cynomolgus CXCR2 expressed in macaque neutrophils. Antibody binding activity was determined as mean fluorescence intensity (MFI) values obtained from 4-8 independent samples.

In fig. 3 shows the results of a typical dose-response study of inhibition of CXCL1-mediated activation of CXCR2 by selected anti-CXCR2 antibodies as measured in Tango™ cells.

In fig. 4A and 4B show an amino acid sequence alignment of representative BKO-4A8 variants with potency similar to the parent BKO-4A8, and the consensus sequence is presented. Fig. 4A = heavy chain variable regions (BKO-4A8 SEQ ID NO: 17; consensus sequence SEQ ID NO: 167); fig. 4B = light chain variable regions (BKO-4A8 SEQ ID NO: 18; consensus sequence SEQ ID NO: 168). The location of the CDR regions in these sequences is given according to Kabat. Accordingly, the 53rd amino acid residue in the alignment in FIG. 4A is numbered 52a according to Kabat (although variant G52aD contains a G to D substitution at residue 53, but the residue is referred to as 52a). Exactly the same in FIG. 4B The 96th amino acid residue is named 95a. In fig. 4A shows SEQ ID NOs: 17, 53-75 and 167 and the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 169-171, respectively, in order of appearance. In fig. 4B shows SEQ ID NOs: 18, 76-97 and 168 and the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 172-174, respectively, in order of appearance.

In fig. 5A and 5B show an alignment of the amino acid sequences of representative combinatorial variants of BKO-4A8 (abbreviated as “Var”) with the parent BKO-4A8 and a consensus sequence based on these variants. Fig. 5A = heavy chain variable regions (BKO-4A8 SEQ ID NO: 17; consensus sequence SEQ ID NO: 226); fig. 5B = light chain variable regions (BKO-4A8 SEQ ID NO: 18; consensus sequence SEQ ID NO: 227). In fig. 5A shows SEQ ID NOs: 17, 98-108, 163-165 and 226 and the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 228-230, respectively, in order of appearance. In fig. 5B shows SEQ ID NOs: 18, 109-110 and 227 and the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 231-232, respectively, in order of appearance.

In fig. 6A and 6B show the results of a dose-response study on the inhibition of CXCL1 (Fig. 6A) and CXCL8 (Fig. 6B)-mediated activation of CXCR2 by selected anti-CXCR2 antibodies BKO-4A8 and optimized antibodies BKO-4A8-101c, BKO-4A8-103c, BKO -4A8-104c and BKO-4A8-105c when measured on Tango™ cells.

In fig. 7 shows the results of a dose-response study on the inhibition of ELR+CXC chemokine-mediated activation of CXCR2 by the indicated anti-CXCR2 antibody BKO-4A8-101c as measured in Tango™ cells.

In fig. 8 shows the results of a dose-response study on the inhibition of CXCL1, CXCL5 and CXCL8-mediated activation of CXCR2 by the indicated anti-CXCR2 antibody BKO-4A8-101c as measured by calcium fluxes.

In fig. 9 shows the CXCR2 binding activity of the indicated anti-CXCR2 antibody BKO-4A8, reformatted on other human IgG constant regions, as determined by flow cytometry.

In fig. 10 shows the results of a dose-response study on the inhibition of CXCL8-mediated activation of CXCR2 by the indicated anti-CXCR2 antibody BKO-4A8 reformatted on other human IgG constant regions, as measured in Tango™ cells.

In fig. 11 shows the results of a binding study of the anti-CXCR2 antibody BKO-4A8-101c with various members of the human CXCR family. The results showed that among human CXCR family members, only BKO-4A8-101c binds to CXCR2.

In fig. 12A and 12B show a representative binding profile of BKO-4A8-101c (shaded histogram) to phenotypically defined human peripheral blood hematopoietic cells as determined by flow cytometry (N=8), including isotype controls (open human IgG histogram). Expression was high in neutrophils (Fig. 12A), whereas intermediate levels of CXCR2 were observed in monocytes (Fig. 12B).

In fig. 13A, 13B and 13C show the results of subcutaneous sensitization and intranasal challenge (on day 14) with house dust mite (HDM) antigen, which caused signs of acute allergic (asthmatic) inflammation in the lungs of human CXCR2 transgenic mice. Specifically, after challenge, mice exhibited moderate to severe multifocal pulmonary inflammation with eosinophils and mild to moderate goblet cell hyperplasia in the bronchioles, compared with control (untreated) mice that had little or no inflammation. Treatment with BKO-4A8-mIgG resulted in a reduction in pathological severity, including a significant reduction in the number of neutrophils (Fig. 13A) and eosinophils in the lungs (Fig. 13B) and mucus density index (Fig. 13C) compared to vehicle.

In fig. 14A and 14B show results of the activity of the anti-CXCR2 antibody BKO-4A8-101c in a cynomolgus monkey model of acute pneumonia. Aerosolized inhalation of lipopolysaccharide (LPS) (on day 0) successfully induced signs of acute neutrophilic inflammation in the lungs of cynomolgus monkeys. Treatment with anti-CXCR2 antibody BKO-4A8-101c (1 mg/kg) 1 hour before LPS administration on day 0 resulted in a significant reduction in the number of neutrophils in bronchoalveolar lavage 24 hours after LPS administration (Fig. 14A). Peripheral blood neutrophil counts were not affected by three repeated doses of BKO-4A8-101c when administered at two-week intervals on days 0, 14, and 28. Medians and intervals by group are presented, N = 4 (FIG. 14B).

In fig. 15A, 15B and 15C show selective inhibition of chemokine-induced CD11b up-regulation in enriched human neutrophils. Antibody against CXCR2 significantly inhibited the response to CXCL1 (p=0.0002) (Fig. 15A) and CXCL5 (p=0.0001) (Fig. 15B). The anti-CXCR2 antibody inhibited significantly more than small molecule antagonist 5 in the same assays (p < 0.0058) (Fig. 15A-B). The CXCL8-mediated increase in CD11b levels was reduced by a CXCR1 antagonist (data not shown) but not by anti-CXCR2 antibody or antagonist 5 (Fig. 15C).

Disclosure of the invention

The disclosed human antibody molecules, methods and uses will become better understood by reference to the following detailed description in conjunction with the accompanying drawings, which are incorporated herein. It should be understood that the disclosed human antibody molecules, methods and uses are not limited to the specific human antibody molecules, methods and uses described and/or provided herein, and the terminology used herein is intended to describe specific embodiments by way of example only and not to limit the claimed molecules human antibodies, methods and applications.

Unless specifically stated otherwise, any description of a possible mechanism or mode of action or reason for improvement is for illustrative purposes only, and the disclosed human antibody molecules, methods and uses are not intended to be limited by the correctness or incorrectness of such proposed mechanism or mode of action or reason for improvement.

Descriptions throughout the text relate to human antibody molecules and methods of using these human antibody molecules. If a feature or embodiment is described or claimed to be associated with human antibody molecules, then such feature or embodiment is equally applicable to methods of using those human antibody molecules. Likewise, if a feature or embodiment is set forth or claimed in connection with a method of using human antibody molecules, then such feature or embodiment is equally applicable to human antibody molecules.

If a range of numeric values is given or specified here, then it includes its endpoints and all the individual integers and fractions within that range, and also includes each of the narrower ranges within it formed by all the different possible combinations of those endpoints and inner integers numbers and fractions, forming subgroups of a larger group of values within a specified range to the same extent as if each of these narrower ranges had been stated directly. If a range of numerical values greater than the specified value is given, the range is nonetheless finite and is limited at its upper end to that value that is valid within the context of the invention as described herein. If a range of numerical values is given that is less than the specified value, then the range is nevertheless limited at its lower end by a value other than zero. The scope of the invention should not be limited to the specific values indicated when defining the range. All ranges are inclusive and can be combined.

It should be understood that certain features of the disclosed human antibody molecules, methods and uses, which for clarity are described herein in the context of individual embodiments, may also be presented in combination as a single embodiment. Conversely, various features of the disclosed human antibody molecules, methods and uses, which for brevity are described in the context of a single embodiment, may also be presented individually or in any subcombination.

Throughout the specification and claims, various terms are used to refer to aspects of the specification. Such terms should be given their art-accepted meaning unless otherwise noted. Other specifically defined terms should be construed in accordance with the definitions provided herein.

In the present invention, singular forms include plural meanings.

The term “about” in relation to numerical ranges, cut-offs or specific values is used to indicate that the values shown may differ by up to 10% from the stated value. Since many of the numerical values used herein are determined experimentally, those skilled in the art will appreciate that such determinations can, and often will, vary from experiment to experiment. The values used here should not be considered overly restrictive due to these inherent variations. Thus, the term “about” generally covers variations of ±10% or less, variations of ±5% or less, variations of ±1% or less, variations of ±0.5% or less, or variations of ±0.1% or less. less than the specified value. When meanings are expressed using the prefix “about,” it is understood that certain forms of meaning constitute other embodiments.

A reference to a particular numerical value includes at least that value itself, unless the context clearly requires otherwise.

The term “including” shall include examples covered by the terms “consisting essentially of” and “consisting of”; likewise, the term “consisting essentially of” must include examples covered by the term “consisting of.”

In the present invention, the expression “wherein the antibody molecule inhibits CXCL1-induced CXCR2 activation or CXCL5-induced CXCR2 activation” and similar expressions refer to the ability of the described human antibody molecules to reduce CXCL1-induced or CXCL5-induced CXCR2 activation, as determined by β-arrestin recruitment on Tango™ cells, by approximately 80%, 85%, 90%, 92%, 95%, 97%, or 100% compared to the level of CXCL1 and/or CXCL5-induced CXCR2 activation in the absence of described human antibody molecules with an IC ₅₀ value of 0 .08 to 0.5 nM at a concentration of 1.5-3.4 nM for CXCL1 and 47.7-150 nM for CXCL5.

In the present invention, “treatment” and similar terms refer to at least one of the following: reducing the severity and/or frequency of symptoms, eliminating symptoms, reducing or eliminating the underlying cause of symptoms, reducing the frequency or likelihood of symptoms and/or their underlying cause, and/or correcting or correction of damage caused directly or indirectly by the diseases or disorders described. Treatment may also include prolonging survival beyond expected survival in subjects not receiving the disclosed human antibody molecules or pharmaceutical compositions containing them.

In the present invention, “prevention” and similar terms refer to preventive or supportive measures. Subjects to receive such preventive or supportive measures are those who are at risk for the described diseases or disorders, for example, due to genetic predisposition or environmental factors, or those who have previously been treated for the described diseases or disorders and are receiving therapeutically effective doses of the described molecules human antibodies or pharmaceutical compositions as a maintenance medication (eg, to maintain low levels of neutrophils in the lungs).

In the present invention, “subject administration” and similar terms mean a procedure in which the described human antibody molecules or pharmaceutical compositions containing them are administered/provided to patients so that target cells, tissues or body segments of the subject are contacted with the described human antibody molecules.

The expression “therapeutically effective amount” means that amount of the described human antibody molecules or pharmaceutical compositions containing them as described herein that is effective to achieve a particular biological, therapeutic or prophylactic result, without limitation, the type of biological or therapeutic results disclosed, described or provided herein . The therapeutically effective amount may vary depending on factors such as the disease, age, sex and weight of the individual, as well as the ability of the composition to produce the desired responses in the subject. Typical indicators of a therapeutically effective amount include, for example, an improvement in the well-being of the subject, a decrease in neutrophilia in one or more peripheral tissues such as a decrease in airway neutrophilia, a decrease in the number of monocytes in one or more peripheral tissues, a decrease in acute airway inflammation, a decrease in chronic airway inflammation, for example, in bronchiectasis, reducing tumor burden, stopping or slowing cancer growth, reducing chronic pain, reducing neuroinflammation of the type seen in multiple sclerosis, reducing inflammation in the liver, reducing inflammation in the pancreas, or reducing symptoms of type I diabetes.

As used herein, “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” means that material which, when combined with an active ingredient (such as disclosed human antibody molecules), allows the ingredient to retain biological activity and does not react with the immune system of a subject. Examples include, but are not limited to, any standard pharmaceutical carriers such as phosphate buffered saline, water, various types of wetting agents (such as Polysorbate 20, Polysorbate 80) and tris(hydroxymethyl)aminomethane (“Tris”) salts such as hydrochloride , acetate, maleate and lactate. Amino acids (such as histidine, glutamine, glutamate, glycine, arginine), sugars (such as sucrose, glucose, trehalose), chelators (such as ETDA) and antioxidants (such as reduced cysteine) can also be added as stabilizing agents. . The preferred diluents for aerosol or parenteral administration are phosphate saline or normal (0.9%) saline. Compositions containing such carriers are formulated by well known standard methods (eg, see Remington's Pharmaceutical Sciences, 18th edition, A. Gennaro, ed., Mack Publishing Co., Easton, PA, 1990; and Remington, The Science and Practice of Pharmacy, 20th Ed., Mack Publishing, 2000). In preferred embodiments, the pharmaceutical compositions are compositions for parenteral administration.

The term “subject” in the present invention refers to cynomolgus monkeys and humans, most preferably humans. In this case, “subject” and “patient” are used in an interdependent manner.

The term “antibody” and similar terms are used in a broad sense to include immunoglobulin molecules, including monoclonal antibodies, antibody fragments, bispecific or multispecific antibodies, dimeric, tetrameric or multimeric antibodies and single chain antibodies. Immunoglobulins can fall into five main classes, namely IgA, IgD, IgE, IgG and IgM, depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3 and IgG4. The light chains of antibodies from any vertebrate species can be classified into one of two clearly distinguishable types, namely kappa (κ) and lambda (λ), based on the amino acid sequences of their constant domains.

“Antibody fragment” means a portion of an immunoglobulin molecule that retains the specific antigen-binding properties of the original full-length antibody (ie, its antigen-binding fragment). Typical antibody fragments contain heavy chain complementarity determining regions (HCDR) 1, 2 and 3 and light chain complementarity determining regions (LCDR) 1, 2 and 3. Other typical antibody fragments contain a heavy chain variable region ( _VH ) and a light chain variable region ( _VL ). Antibody fragments include, but are not limited to: Fab fragment, a monovalent fragment consisting of V _L , V _H , light chain ( _CL ) and heavy chain 1 ( _CH 1 ) constant domains; F(ab) ₂ fragment, a bivalent fragment containing two Fab fragments connected by a disulfide bridge in the hinge region; and an Fv fragment consisting of the V _L and V _H domains of one arm of the antibody. The V _H and V _L domains can be engineered and linked together via a synthetic linker to form various types of single chain antibody constructs in which the V _H /V _L domains form intramolecular or intermolecular pairs when the V _H and V _L domains are expressed separate single-chain antibody constructs to form a monovalent antigen-binding site, such as single-chain Fv (scFv) or diabodies; as described, for example, in Int. Pat. Pub. Nos. WO 1998/044001, WO 1988/001649, WO 1994/013804 and WO 1992/001047. These antibody fragments are prepared by methods well known to those skilled in the art, and the fragments are tested for suitability in the same manner as full-length antibodies.

Each variable region of an antibody heavy chain or light chain consists of four “framework” regions (FR), which alternate with three “complementarity determining regions” (CDR) in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (from N - to the C-terminus). The three CDRs in V _H are designated as HCDR1, HCDR2, HCDR3, and the three CDRs in V _L are designated as LCDR1, LCDR2, LCDR3, respectively. The location and size of CDRs are determined by rules that identify areas of sequence variation in immunoglobulin variable regions (Wu and Kabat, J Exp Med 132:211-50, 1970; Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service , National Institutes of Health, Bethesda, Md., 1991). Amino acid residues of the variable region are numbered according to the Kabat (ibid.) system. “Framework regions” are the remaining sequences of the variable region other than those defined as CDRs.

“Human antibody” means an antibody containing heavy and light chain variable regions in which both framework regions and antigen binding sites are derived from sequences of human origin. If the antibody contains a constant region, then the constant region is also derived from sequences of human origin. A human antibody contains heavy or light chain variable regions that are “derived from” sequences of human origin if the antibody variable regions are derived from a system that uses human germline immunoglobulin genes or rearranged human immunoglobulin genes. Such systems include libraries of human immunoglobulin genes in phage display systems and transgenic animals such as mice or rats carrying human immunoglobulin loci. A “human antibody” may contain amino acid differences compared to human germline immunoglobulin sequences or rearranged immunoglobulin sequences, for example due to naturally occurring somatic mutations or deliberate introduction of substitutions into framework regions or antigen binding sites. Typically, a “human antibody” in amino acid sequence is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence encoded by human germline immunoglobulin sequences or the rearranged human immunoglobulin gene. In some cases, the “human antibody” may contain consensus framework sequences derived from analysis of human frameworks, for example, as described in Knappik et al., J Mol Biol 296: 57-86, 2000, or synthetic HCDR3 included in a gene library human immunoglobulin in a phage display system, as described, for example, in Shi et al., J Mol Biol 397:385-96, 2010; and Int. Pat. Pub. No. WO 2009/085462.

Human antibodies, although derived from human immunoglobulin sequences, can be produced in phage display type systems incorporating synthetic CDRs and/or synthetic framework regions and/or can be mutagenized in vitro to improve antibody properties in the variable regions or constant regions or both by producing antibodies that do not naturally exist in the human germline antibody repertoire in vivo.

“Monoclonal antibody” means a population of antibody molecules with essentially the same molecular composition. A monoclonal antibody exhibits only one binding specificity and affinity for a particular epitope, and in the case of a bispecific monoclonal antibody, dual binding specificity for two different epitopes. Thus, a monoclonal antibody means a population of antibodies with a single amino acid composition in each heavy and each light chain, excluding possible well-known variations such as removal of the C-terminal lysine from the heavy chain of the antibody and processing variants in which incomplete cleavage of the N-terminal leader sequence occurs , which is produced in the cell and is usually broken off during secretion. For example, US Pat. No. 8,241,630 describes a commercial antibody in which 5-15% of the antibody population retains the leader sequence. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. A monoclonal antibody may be monospecific or multispecific, or monovalent, bivalent or polyvalent. The term monoclonal antibody also includes bispecific antibodies.

“Epitope” means that portion of an antigen to which an antibody specifically binds. Epitopes typically consist of chemically active (such as polar, nonpolar, or hydrophobic) surface moieties of molecules such as amino acids or polysaccharide side chains, and may have specific three-dimensional structural characteristics as well as specific charge characteristics. An epitope may consist of contiguous and/or non-contiguous amino acids forming a conformational spatial unit. In a non-contiguous epitope, amino acids from different parts of the linear antigen sequence come into close proximity in three-dimensional space due to the folding of the protein molecule.

“Variant” means a polypeptide or polynucleotide that differs from a reference polypeptide or reference polynucleotide by one or more modifications, for example, substitutions, insertions or deletions.

The expression “immunospecifically binds” refers to the ability of the described human antibody molecules to preferentially bind to their target (CXCR2 in the case of anti-CXCR2 antibodies) without preferential binding to other molecules of the CXCR family in samples containing mixed populations of molecules. Human antibody molecules that immunospecifically bind to CXCR2 contain virtually no other antibodies that have different antigen specificities (eg, anti-CXCR2 antibody contains virtually no antibodies that specifically bind antigens other than CXCR2). However, antibody molecules that immunospecifically bind to human CXCR2 may be cross-reactive to other antigens such as human CXCR2 orthologs, including Macaca fascicularis CXCR2. The antibody molecules described herein are capable of immunospecifically binding to both native human CXCR2 and human CXCR2 that is produced recombinantly in mammalian cells or prokaryotic cells.

In the present invention, “severe persistent neutropenia” means that the absolute neutrophil count (ANC) in the peripheral blood is less than 0.4×10 ⁹ cells/L for more than 2 weeks. Severe persistent neutropenia can be classified as follows:

• Grade 1 means mild case (0.8-1.0×10 ⁹ cells/l)

• Grade 2 means moderate case (0.6-0.8×10 ⁹ cells/l)

• Grade 3 means severe case (0.4-0.6×10 ⁹ cells/l)

• Grade 4 means potentially life-threatening (less than 0.4x10 ⁹ cells/l)

(See Table for Adverse Event Severity Classification in Adults and Children from the Division of AIDS (DAIDS), National Institute of Allergy and Infectious Diseases, National Institutes of Health, US Department of Health and Human Services, version 2.0, November 2014).

In the present invention, the following abbreviations are used: heavy chain variable region ( _VH ); light chain variable region ( _VL ); complementarity determining region (CDR); heavy chain CDR (HCDR); light chain CDR (LCDR); CXC chemokine receptor-2 (CXCR2); and CXC chemokine ligand 1, 2, 3, 5, 6, 7, and 8 (CXCL1, 2, 3, 5, 6, 7, and 8).

The disclosed antibody molecules may contain one or more substitutions, deletions or insertions in framework regions and/or constant regions. In some embodiments, IgG4 antibody molecules may contain the S228P substitution. S228 (EU residue numbering) is located in the hinge region of IgG4 antibody molecules. The replacement of serine (“S”) with proline (“P”) serves to stabilize the IgG4 hinge and prevent Fab arm turnover in vitro and in vivo. In some embodiments, the antibody molecules may contain one or more modifications that increase the half-life of the antibody molecules in vivo. For example, in some embodiments, antibodies may contain the M252Y substitution, the S254T substitution, and the T256E substitution (collectively referred to as the “YTE” substitution). M252, S254 and T256 (EU residue numbering) are located in the C _H 2 domain of the heavy chain. Replacement of these residues with tyrosine (“Y”), threonine (“T”), and glutamate (“E”), respectively, protects antibody molecules from lysosomal degradation, thereby increasing the half-life of antibody molecules in serum. In some embodiments, the antibody molecules may contain a deletion of the C-terminal heavy chain lysine residue. Deletion of the C-terminal heavy chain lysine residue reduces the heterogeneity of antibody molecules produced by mammalian cells. In some embodiments, antibody molecules may contain a combination of substitutions, deletions, or insertions. For example, in some aspects, the disclosed antibody molecules may comprise an S228P substitution and a deletion of a C-terminal heavy chain lysine residue. Constant regions of various classes of antibodies are known to be involved in modulating antibody effector functions such as antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and antibody-dependent phagocytosis (ADP). In some embodiments, the described antibody molecules may contain one or more substitutions, deletions or insertions in constant regions that modulate one or more effector functions of the antibodies, such as reducing or eliminating one or more effector functions. Other changes are known that affect the effector functions of antibodies and the half-life in the bloodstream. For example, see Saunders KO “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life” Front. Immunol. (2019) 10:1296.

Human antibody molecules

Human antibody molecules are provided that bind immunospecifically to human CXCR2. The human antibody molecules may contain the heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 167 and the light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 168 and inhibit the activation of CXCR2 by CXCL1 or CXCL5. Given in table. 19 and FIG. 4A and 4B, SEQ ID NO: 167 and 168 represent the consensus heavy chain variable region and the light chain variable region, respectively (SEQ ID NO: 167 = “consensus _VH ” and SEQ ID NO: 168 = “consensus _VL ”), described human antibody molecules. The consensus CDR sequences are provided as SEQ ID NOs: 169, 170, 171, 172, 173 and 174. Numbering in the names of these CDR sequences, unless otherwise noted, is according to Kabat. In some embodiments, human antibody molecules may include:

Heavy chain CDR1 having the amino acid sequence SX ₁ X ₂ X ₃ S, where X ₁ is S, Q, H, L, W or Y; X ₂ - T or A; and X ₃ is M, Q, D, H or W as set forth in SEQ ID NO: 169;

Heavy chain CDR2 having the amino acid sequence AX ₄ SX ₅ X ₆ X ₇ RX ₈ TYYADSVKG, where X ₄ is I or H; X ₅ - G or D; X ₆ means R, S or Q; X ₇ - G or D; and X ₈ is N or S as set forth in SEQ ID NO: 170;

A heavy chain CDR3 having the amino acid sequence QX ₁₀ X ₁₁ X ₁₂ , where X ₁₀ is M, A, Q or K; X ₁₁ - G or D; and X ₁₂ is Y, S or K as set forth in SEQ ID NO: 171;

Light chain CDR1 having the amino acid sequence IGTSSDVGGYNYVS as set forth in SEQ ID NO: 172;

A light chain CDR2 having the amino acid sequence X ₁₃ VX ₁₄ X ₁₅ X ₁₆ PS, where X ₁₃ is E or D; X ₁₄ - N, D or S; X ₁₅ means K, A, D or H; and X ₁₆ is R or Q as set forth in SEQ ID NO: 173; And

A light chain CDR3 having the amino acid sequence SSX ₁₇ AGX ₁₈ NX ₁₉ FGX ₂₀ , where X ₁₇ is Y or A; X ₁₈ - N, A, S, K, L, W or Y; X ₁₉ means N, Q, D, H, K, L or Y; and X ₂₀ is V, A, or K as set forth in SEQ ID NO: 174.

Human antibody molecules may contain CDR1, CDR2 and CDR3 of the heavy chain according to SEQ ID NO: 226 and CDR1, CDR2 and CDR3 of the light chain according to SEQ ID NO: 227. Shown in table. 19 and FIG. 5A and 5B, SEQ ID NO: 226 and 227 represent the consensus heavy chain variable region and the light chain variable region, respectively (SEQ ID NO: 226 = “consensus _VH ” and SEQ ID NO: 227 = “consensus _VL ”). described human antibody molecules. The consensus CDR sequences are provided as SEQ ID NOs: 228, 229, 230, 201, 231 and 232. In some embodiments, human antibody molecules may include:

A heavy chain CDR1 having the amino acid sequence SSTX ₂₁ S, where X ₂₁ is M or Q, as shown in SEQ ID NO: 228;

Heavy chain CDR2 having the amino acid sequence AISGX ₂₃ GX ₂₄ X ₂₅ TYYADSVKG, where X ₂₃ is R or S; X ₂₄ - R or G; X ₂₅ means N or S, as presented in SEQ ID NO: 229;

A heavy chain CDR3 having the amino acid sequence QX ₂₈ GY, wherein X ₂₈ is M, K or A as set forth in SEQ ID NO: 230;

A light chain CDR1 having the amino acid sequence IGTSSDVGGY‌NYVS as set forth in SEQ ID NO: 201;

A light chain CDR2 having the amino acid sequence EVX ₃₀ KRPS, where X ₃₀ is N or S, as shown in SEQ ID NO: 231; And

A light chain CDR3 having the amino acid sequence SSYAGX ₃₁ NN FGV, where X ₃₁ is N or S, as set forth in SEQ ID NO: 232.

The described human antibody molecules may contain combinations of heavy chain and light chain CDRs presented in table. 1. In some embodiments, for example, human antibody molecules may comprise a heavy chain CDR1 having the amino acid sequence of any of SEQ ID NOs: 175-185, a heavy chain CDR2 having the amino acid sequence of any of SEQ ID NOs: 186-192, Heavy chain CDR3 having the amino acid sequence of any of SEQ ID NO: 194-200, Light chain CDR1 having the amino acid sequence of SEQ ID NO: 201, Light chain CDR2 having the amino acid sequence of any of SEQ ID NO: 202-209, and a light chain CDR3 having the amino acid sequence of any one of SEQ ID NO: 210-225.

Table 1. Heavy chain and light chain CDR sequences

Antibody chain with position substitution according to Kabat SEQ ID NO: CDR1 CDR2 CDR3 Heavy chain (HC) variable region V _H S32Q 4A8 SEQ ID NO: 53 SEQ ID NO: 176 SEQ ID NO: 186 SEQ ID NO: 194 V _H S32H 4A8 SEQ ID NO: 54 SEQ ID NO: 177 SEQ ID NO: 186 SEQ ID NO: 194 V _H S32L 4A8 SEQ ID NO: 55 SEQ ID NO: 178 SEQ ID NO: 186 SEQ ID NO: 194 V _H S32W 4A8 SEQ ID NO: 56 SEQ ID NO: 179 SEQ ID NO: 186 SEQ ID NO: 194 V _H S32Y 4A8 SEQ ID NO: 57 SEQ ID NO: 180 SEQ ID NO: 186 SEQ ID NO: 194 V _H T33A 4A8 SEQ ID NO: 58 SEQ ID NO: 181 SEQ ID NO: 186 SEQ ID NO: 194 V _H M34Q 4A8 SEQ ID NO: 59 SEQ ID NO: 182 SEQ ID NO: 186 SEQ ID NO: 194 V _H M34D 4A8
SEQ ID NO: 60 SEQ ID NO: 183 SEQ ID NO: 186 SEQ ID NO: 194 V _H M34H 4A8
SEQ ID NO: 61 SEQ ID NO: 184 SEQ ID NO: 186 SEQ ID NO: 194 V _H M34W 4A8
SEQ ID NO: 62 SEQ ID NO: 185 SEQ ID NO: 186 SEQ ID NO: 194 V _H I51H 4A8
SEQ ID NO: 63 SEQ ID NO: 175 SEQ ID NO: 187 SEQ ID NO: 194 V _H G52aD 4A8
SEQ ID NO: 64 SEQ ID NO: 175 SEQ ID NO: 188 SEQ ID NO: 194 V _H R53S 4A8
SEQ ID NO: 65 SEQ ID NO: 175 SEQ ID NO: 189 SEQ ID NO: 194 V _H R53Q 4A8
SEQ ID NO: 66 SEQ ID NO: 175 SEQ ID NO: 190 SEQ ID NO: 194 _VH G54D 4A8
SEQ ID NO: 67 SEQ ID NO: 175 SEQ ID NO: 191 SEQ ID NO: 194 V _H N56S 4A8
SEQ ID NO: 68 SEQ ID NO: 175 SEQ ID NO: 192 SEQ ID NO: 194 V _H M96A 4A8
SEQ ID NO: 70 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 195 V _H M96Q 4A8
SEQ ID NO: 71 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 196 V _H M96K 4A8
SEQ ID NO: 72 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 197 V _H G101D 4A8
SEQ ID NO: 73 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 198 V _H Y102S 4A8
SEQ ID NO: 74 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 199 V _H Y102K 4A8
SEQ ID NO: 75 SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 200 V _H M34Q, N56S 4A8
(SEQ ID NO: 98) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 194 V _H M34Q, A40P, N56S 4A8 (SEQ ID NO: 99) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 194 V _H M34Q, A40P, N56S, R75K 4A8 (SEQ ID NO: 100) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 194 V _H M34Q, A40P, N56S, M96K 4A8 (SEQ ID NO: 101) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H M34Q, A40P, N56S, R75K, M96K 4A8
(SEQ ID NO: 102) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H M34Q, A40P, N56S, R75K, I94K, M96K 4A8
(SEQ ID NO: 103) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H M34Q, A40P, N56S, I94K, M96K 4A8
(SEQ ID NO: 104) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H M34Q, N56S, M96K 4A8
(SEQ ID NO: 105) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H M34Q, N56S, R75K, M96K 4A8 (SEQ ID NO: 106) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 197 V _H I94K, M96K 4A8
(SEQ ID NO: 107) SEQ ID NO: 175 SEQ ID NO: 186 SEQ ID NO: 197 V _H M34Q, A40P, N56S, R75K, M96A 4A8
(SEQ ID NO: 108) SEQ ID NO: 182 SEQ ID NO: 192 SEQ ID NO: 195 Light chain (LC) variable region V _L E50D 4A8
SEQ ID NO: 76 SEQ ID NO: 201 SEQ ID NO: 203 SEQ ID NO: 210 V _L N52D 4A8
SEQ ID NO: 77 SEQ ID NO: 201 SEQ ID NO: 204 SEQ ID NO: 210 V _L N52S 4A8
SEQ ID NO: 78 SEQ ID NO: 201 SEQ ID NO: 205 SEQ ID NO: 210 V _L K53A 4A8
SEQ ID NO: 79 SEQ ID NO: 201 SEQ ID NO: 206 SEQ ID NO: 210 V _L K53D 4A8
SEQ ID NO: 80 SEQ ID NO: 201 SEQ ID NO: 207 SEQ ID NO: 210 V _L K53H 4A8
SEQ ID NO: 81 SEQ ID NO: 201 SEQ ID NO: 208 SEQ ID NO: 210 V _L R54Q 4A8
SEQ ID NO: 82 SEQ ID NO: 201 SEQ ID NO: 209 SEQ ID NO: 210 V _L Y91A 4A8
SEQ ID NO: 83 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 211 V _L N94A 4A8
SEQ ID NO: 84 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 212 V _L N94S 4A8
SEQ ID NO: 85 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 213 V _L N94K 4A8
SEQ ID NO: 86 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 214 V _L N94L 4A8
SEQ ID NO: 87 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 215 V _L N94W 4A8
SEQ ID NO: 88 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 216 V _L N94Y 4A8
SEQ ID NO: 89 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 217 V _L N95aQ 4A8
SEQ ID NO: 90 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 218 V _L N95aD 4A8
SEQ ID NO: 91 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 219 V _L N95aH 4A8
SEQ ID NO: 92 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 220 V _L N95aK 4A8
SEQ ID NO: 93 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 221 V _L N95aL 4A8
SEQ ID NO: 94 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 222 V _L N95aY 4A8
SEQ ID NO: 95 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 223 V _L V97A 4A8
SEQ ID NO: 96 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 224 V _L V97K 4A8
SEQ ID NO: 97 SEQ ID NO: 201 SEQ ID NO: 202 SEQ ID NO: 225 V _L N52S, N94S 4A8
(SEQ ID NO: 109) SEQ ID NO: 201 SEQ ID NO: 205 SEQ ID NO: 213 V _L D41G, N52S, N94S 4A8
(SEQ ID NO: 110) SEQ ID NO: 201 SEQ ID NO: 205 SEQ ID NO: 213

Residue substitution positions were determined according to Kabat.

In some aspects, human antibody molecules may comprise:

CDR1, CDR2 and CDR3 of the heavy chain

SEQ ID NO: 176, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 177, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 178, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 179, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 180, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 181, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 182, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 183, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 184, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 185, SEQ ID NO: 186 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 187 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 188 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 189 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 190 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 191 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 192 and SEQ ID NO: 194, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 195, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 196, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 197, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 198, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 199, respectively; or

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 200, respectively; And

Light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 210, respectively.

In some aspects, human antibody molecules may comprise:

Heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 194, respectively, and

CDR1, CDR2 and CDR3 of the light chain

SEQ ID NO: 201, SEQ ID NO: 203 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 204 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 205 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 206 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 207 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 208 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 209 and SEQ ID NO: 210, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 211, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 212, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 213, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 214, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 215, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 216, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 217, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 218, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 219, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 220, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 221, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 222, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 223, respectively;

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 224, respectively; or

SEQ ID NO: 201, SEQ ID NO: 202 and SEQ ID NO: 225, respectively.

In some aspects, human antibody molecules may comprise:

CDR1, CDR2 and CDR3 of the heavy chain

SEQ ID NO: 182, SEQ ID NO: 192 and SEQ ID NO: 194, respectively;

SEQ ID NO: 182, SEQ ID NO: 192 and SEQ ID NO: 197, respectively;

SEQ ID NO: 175, SEQ ID NO: 186 and SEQ ID NO: 197, respectively; or

SEQ ID NO: 182, SEQ ID NO: 192 and SEQ ID NO: 195, respectively; And

Light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 201, SEQ ID NO: 205, and SEQ ID NO: 213, respectively.

Human antibody molecules may comprise a heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 182, a heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 192, a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 195, a light chain CDR1 having the amino acid sequence of SEQ ID NO: 201, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 205, and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 213.

Given in table. 19 and FIG. 4A and 4B, SEQ ID NO: 167 and 168 represent the consensus heavy chain variable region and the light chain variable region, respectively (SEQ ID NO: 167 = “consensus _VH ” and SEQ ID NO: 168 = “consensus _VL ”), described human antibody molecules. Thus, the described human antibody molecules may contain a heavy chain variable region having the amino acid sequence EVQLLESGGGLVQPGGSLRLSCAASGTFFS WVRQAPGKGLEWVS RFTISRDNSRNTLYLQMNSLRAEDTAVYYCAX ₉ WGQGILVTVSS, where X ₁ means S, Q, H, L, W or Y; X ₂ - T or A; X ₃ means M, Q, D, H or W; X ₄ - I or H; X ₅ - G or D; X ₆ means R, S or Q; X ₇ - G or D; X ₈ - N or S; X ₉ - I or K; X ₁₀ means M, A, Q or K; X ₁₁ - G or D; and X ₁₂ is Y, S or K as set forth in SEQ ID NO: 167, and a light chain variable region having the amino acid sequence QSALTQPPSASGSPGQSVTISC GVPDRFSGSKSGNTASLTVSGLQAEDEADYYC where X ₁₃ means E or D; X ₁₄ - N, D or S; X ₁₅ means K, A, D or H; X ₁₆ - R or Q; X ₁₇ - Y or A; X ₁₈ means N, A, S, K, L, W or Y; X ₁₉ means N, Q, D, H, K, L or Y; and X ₂₀ is V, A or K, as shown in SEQ ID NO: 168. The underlined residues represent consensus CDRs, as described above.

Human antibody molecules may contain a heavy chain variable region having the amino acid sequence

where X ₂₁ means M or Q; X ₂₂ - A or P; X ₂₃ means R or S; X ₂₄ - R or G; X ₂₅ means N or S; X ₂₆ - R or K; X ₂₇ means I or K; and X ₂₈ is M, K or A as set forth in SEQ ID NO: 226, and a light chain variable region having the amino acid sequence QSALTQPPSASGSPGQSVTISC GVPDRFSGSKSGNTASLTVSGLQAEDEADYYC FGGGTKLTVL, where X ₂₉ means D or G; X ₃₀ - N or S; and X ₃₁ is N or S as set forth in SEQ ID NO: 227. The underlined residues represent consensus CDRs as described above.

Human antibody molecules may include:

a) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 98, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

b) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 99, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

c) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 100, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

d) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 101, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

e) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 102, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

f) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 103, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

g) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 104, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

h) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 105, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

i) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 106, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

j) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 107, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

k) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 108, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

l) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 162, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

m) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 163, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

n) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 164, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110;

o) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 165, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110; or

p) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 166, and a light chain variable region having the amino acid sequence of SEQ ID NO: 109 or 110.

In some embodiments, the human antibody molecules include:

a) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 108, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110;

b) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 162, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110;

c) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 163, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110;

d) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 164, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110;

e) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 165, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110; or

f) a heavy chain variable region having the amino acid sequence of SEQ ID NO: 166, and a light chain variable region having the amino acid sequence of SEQ ID NO: 110.

Human antibody molecules may include a heavy chain variable region having the amino acid sequence of SEQ ID NO: 108 and a light chain variable region having the amino acid sequence of SEQ ID NO: 110.

The described human antibody molecules may contain a human IgG1, IgG2 or IgG4 heavy chain constant region. In some embodiments, the human antibody molecules comprise a human IgG1 heavy chain constant region. Suitable human IgG1 heavy chain constant regions include, for example, the amino acid sequence of SEQ ID NO: 122 or 124. In some aspects, the human IgG1 heavy chain constant region includes the amino acid sequence of SEQ ID NO: 122. In some aspects, the human IgG1 heavy chain constant region includes the amino acid sequence of SEQ ID NO: 122. In some aspects, the IgG1 heavy chain constant region human comprises the amino acid sequence of SEQ ID NO: 124. In some embodiments, the human antibody molecules comprise a human IgG2 heavy chain constant region. In some aspects, the human IgG2 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 120. In some embodiments, the human antibody molecules comprise a human IgG4 heavy chain constant region. Suitable human IgG4 heavy chain constant regions include, for example, the amino acid sequence of SEQ ID NO: 116 or 118. In some embodiments, the human IgG4 heavy chain constant region includes the amino acid sequence of SEQ ID NO: 116. In some aspects, the human IgG4 heavy chain constant region includes the amino acid sequence of SEQ ID NO: 116. In some aspects, the IgG4 heavy chain constant region human comprises the amino acid sequence of SEQ ID NO: 118. In some embodiments, the human IgG4 heavy chain constant region contains the S228P substitution. In some embodiments, the human IgG4 heavy chain constant region contains substitutions M252Y, S254T, and T256E. In some embodiments, the human IgG4 heavy chain constant region contains a deletion of the C-terminal lysine residue relative to wild-type IgG4.

Human antibody molecules may contain a human lambda light chain constant region (λ) or a human kappa light chain constant region (κ). In some embodiments, the human antibody molecules comprise a human lambda II light chain constant region (λ2).

Human antibody molecules may contain a human IgG1 heavy chain constant region and a human lambda II light chain constant region. Human antibody molecules may contain a human IgG2 heavy chain constant region and a human lambda II light chain constant region. Human antibody molecules may contain a human IgG4 heavy chain constant region and a human lambda II light chain constant region.

Human antibody molecules can be full-length antibodies or antigen-binding fragments thereof. Suitable antigen binding fragments include, for example, Fab fragments, F(ab) ₂ fragments or single chain antibodies.

The described human antibody molecules are selective antagonists of human CXCR2, thereby inhibiting CXCR2 activation induced by CXCL1 or CXCL5. The described human antibody molecules can also partially inhibit CXCL8-induced CXCR2 activation. The described human antibody molecules may also exhibit one or more of the following properties:

a) inhibit CXCL1-induced calcium influx in HTS002C-Chemiscreen™ cells with calcium-optimized chemokine receptor CXCR2 with an IC ₅₀ value of 0.8 to 2.4 nM at a CXCL1 concentration of 1.5 to 3.4 nM;

b) virtually no inhibition of CXCL8-induced calcium influx in HTS002C-Chemiscreen™ cells with the calcium-optimized chemokine receptor CXCR2;

c) inhibit CXCL1 or CXCL5 induced β-arrestin recruitment in Tango™ cell assays with an _IC50 value of 0.08 to 0.5 nM at concentrations of 1.5 to 3.4 nM for CXCL1 and 47.7 to 150 nM for CXCL5; or

d) reduce airway neutrophilia in subjects with airway neutrophilia without causing severe persistent neutropenia.

Pharmaceutical compositions containing any of the human antibody molecules described herein are also provided. In some embodiments, the pharmaceutical compositions may contain any of the human antibody molecules described herein in combination with a pharmaceutically acceptable carrier.

Nucleic acid molecules encoding any of the human antibody molecules described herein are also provided. Exemplary polynucleotides encoding human antibodies or fragments thereof as described herein are provided as SEQ ID NOs: 233-247. Exemplary polynucleotides encoding the heavy chain constant regions of human antibodies are shown as SEQ ID NO: 248-256.

Vectors containing the nucleic acid molecules described herein are also provided.

Cells transformed to express any of the human antibody molecules described herein are also provided.

Methods of treatment and application

CXCR2 antagonists have been the subject of research, including clinical trials, in a range of diseases, including pathologies associated with neutrophilic and/or monocytic inflammation, and certain cancers that express CXCR2 or have elements of neutrophil suppression of anticancer responses. Small molecule antagonists of CXCR2 have been developed, for example from:

(a) COPD (for example, see Miller B. et al. (2017) Late Breaking Abstract - “Danirixin (GSK1325756) improves respiratory symptoms and health status in mild to moderate COPD - results of a 1 year first time in patient study .” European Respiratory Journal, 50);

(b) influenza (for example, see study NCT02469298, described on ClinicalTrials.gov under the title “Safety, Tolerability and Clinical Effect of Danirixin in Adults With Influenza”);

(c) bronchiectasis (for example, see De Soyza et al. (2015) “A randomized, placebo-controlled study of the CXCR2 antagonist AZD5069 in bronchiectasis” Eur. Respir. J., 46, 1021-32);

(d) cystic fibrosis (for example, see Moss et al. (2013) “Safety and early treatment effects of the CXCR2 antagonist SB-656933 in patients with cystic fibrosis” J. Cyst. Fibros., 12, 241-8);

(e) severe asthma (for example, see Nair et al. (2012) “Safety and efficacy of a CXCR2 antagonist in patients with severe asthma and sputum neutrophils: a randomized, placebo-controlled clinical trial” Clin. Exp. Allergy, 42, 1097-103); And

(f) prostate cancer (for example, see study NCT03177187, described on ClinicalTrials.gov under the title “Combination Study of AZD5069 and Enzalutamide”).

In addition, there is evidence that CXCR2 antagonism may be beneficial in chronic upper respiratory diseases such as chronic rhinosinusitis (for example, see Tomassen et al. (2016) “Inflammatory endotypes of chronic rhinosinusitis based on cluster analysis of biomarkers” J Allergy Clin. Immunol., 137, 1449-1456 e4); for vascular diseases, including ischemia-reperfusion injury (Stadtmann and Zarbock (2012) “CXCR2: From Bench to Bedside” Front Immunol., 3, 263) and coronary heart disease (for example, see Joseph et al. (2017) “CXCR2 inhibition - a no-vel approach to treating coronary heart disease (CICADA): study protocol for a randomized controlled trial” Trials, 18, 473); for chronic pain (for example, see Silva et al. (2017) “CXCL1/CXCR2 signaling in pathological pain: Role in peripheral and central sensitization” Neurobiol. Dis., 105, 109-116); in neuroinflammatory diseases (for example, see Veenstra and Ransohoff (2012) “Chemokine receptor CXCR2: physiology regulator and neuroinflammation controller?” J. Neuroimmunol., 246, 1-9), including multiple sclerosis (for example, see Pierson et al (2018) “The contribution of neutrophils to CNS autoimmunity” Clin. Immunol., 189, 23-28) and Alzheimer's disease (for example, see Liu et al. (2014) “Neuroinflammation in Alzheimer's disease: chemokines produced by astrocytes and chemokine receptors" Int. J. Clin. Exp. Pathol., 7, 8342-55); in alcoholic and non-alcoholic steatohepatitis (for example, see French et al. (2017) “The role of the IL-8 signaling pathway in the infiltration of granulocytes into the livers of patients with alcoholic hepatitis” Exp. Mol. Pathol., 103 , 137-140; and Ye et al. (2016) “Lipocalin-2 mediates non-alcoholic steatohepatitis by promoting neutrophil-macrophage crosstalk via the induction of CXCR2” J. Hepatol., 65, 988-997); for pancreatitis (for example, see Steele et al. (2015) “CXCR2 inhibition suppresses acute and chronic pancreatic inflammation” J. Pathol., 237, 85-97); for diabetes (for example, see Citro et al. (2015) “CXCR1/2 inhibition blocks and reverses type 1 diabetes in mice” Diabetes, 64, 1329-40); and in many types of cancer (for example, see Liu et al. (2016) “The CXCL8-CXCR1/2 pathways in cancer” Cytokine Growth Factor Rev., 31, 61-71). Behcet's disease is characterized by activation of neutrophils and has also been associated with CXCR2 (Qiao et al. “CXCR2 expression on neutrophils is upregulated during the relapsing phase of ocular Behcet disease” Curr. Eye Res. 2005, 30:195-203).

The methods include administering to subjects a therapeutically effective amount of any of the human antibody molecules described herein or the pharmaceutical compositions described herein for the treatment or prevention of the inflammatory diseases described herein. In some embodiments, human antibody molecules or pharmaceutical compositions containing them are administered in a therapeutically effective amount to treat airway neutrophilia, as measured by, for example, the number of neutrophils in sputum, or acute pneumonia. In such embodiments, subjects receiving human antibody molecules or pharmaceutical compositions containing them have airway neutrophilia or acute pneumonia. In some embodiments, human antibody molecules or pharmaceutical compositions containing them are administered in a therapeutically effective amount for the prevention of airway neutrophilia or acute pneumonia. In such embodiments, subjects receiving human antibody molecules or pharmaceutical compositions containing them are at risk of developing airway neutrophilia or acute pneumonia, for example, due to genetic predisposition or environmental factors, or have previously been treated for airway neutrophilia or acute inflammation lungs and are receiving or should receive therapeutically effective doses of the described human antibody molecules or pharmaceutical compositions as maintenance medication (eg, to maintain low neutrophil levels in the lungs).

The described human antibody molecules or the described pharmaceutical compositions are also provided for use in the prevention or treatment of respiratory neutrophilia or acute pneumonia, and the use of any of the described human antibody molecules or any of the described pharmaceutical compositions in the manufacture of medicaments for the prevention or treatment of neutrophilia in peripheral tissues, airway neutrophilia, or acute or chronic pneumonia.

Airway neutrophilia, acute pneumonia, or both may present as chronic obstructive pulmonary disease, severe neutrophilic asthma, or both.

The antibodies described here have been shown to inhibit eosinophil migration into the lung in response to inflammatory stimuli in at least one experimental model. Accordingly, these antibodies are useful for the treatment of inflammatory diseases characterized by eosinophilia, such as eosinophilic asthma, allergic rhinitis, skin diseases, fungal and parasitic infections, autoimmune diseases (such as inflammatory bowel disease, neuromyelitis optica, bullous pemphigoid, autoimmune myocarditis, primary biliary liver cirrhosis, eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome), some types of cancer and bone marrow diseases.

Without being bound by any theory, the ability of the present anti-CXCR2 antibodies to block the migration of eosinophils into the lungs is unexpected since eosinophils lack the CXCR2 receptor. In order for inflammatory cells to migrate to a site, they must first cross the lining of blood vessels, which is made up of endothelial cells, which are known to express CXCR2. Accordingly, antibodies are also capable of inhibiting cell migration by affecting endothelial cells.

The ability to target endothelial cells also suggests that the present antibodies may influence angiogenesis and metastasis, which is important in cancer. Accordingly, a method for treating cancer is provided herein.

EXAMPLES

The following examples are provided to further describe some of the embodiments described herein. The examples are intended to be illustrative and not limiting of the embodiments described.

General methods

Creation of plasmids for antibody production

The amino acid sequences of the variable regions were reverse translated into DNA sequences prior to synthesis of the resulting de novo DNA. The synthesized heavy chain variable region genes were subcloned into an expression vector containing a polynucleotide sequence encoding a human IgG4 constant region containing the S228P substitution stabilizing the hinge region (SEQ ID NO: 118). The synthesized lambda light chain variable region genes were subcloned into an expression vector containing a polynucleotide sequence encoding the amino acid sequence of the human lambda light chain constant region (SEQ ID NO: 134). The synthesized kappa light chain variable region genes were subcloned into an expression vector containing a polynucleotide sequence encoding the amino acid sequence of the human kappa light chain constant region (SEQ ID NO: 135).

Short-term antibody expression using the Expi293F™ system

Antibodies were generated by cotransfecting plasmids for the antibody heavy and light chains into Expi293™ cells (Life Technologies). For every 20 ml of transfection, 3.6 x 10 ⁷ cells in 20 ml of Expi293™ expression medium were required. Transfection was performed using ExpiFectamine™ 293 reagent according to the manufacturer's instructions.

Antibodies were collected by centrifugation (3000 × g for 20 min) between 72 and 84 h after transfection. Unless otherwise stated, all antibodies were generated as human IgG4 incorporating the S228P substitution stabilizing the hinge region.

Antibody Purification and Buffer Exchange

Antibodies were purified from short-term transfection material using protein A resin (MabSelect™ SuRe™, GE Healthcare) according to the manufacturer's instructions.

After elution of the antibodies, the buffer was replaced from citric acid with PBS according to Sorensen, pH 5.8 (59.5 mM KH ₂ PO ₄ , 7.3 mM Na ₂ HPO ₄ , 2H ₂ 0, 150 mM NaCl), using desalting columns PD-10 (52-1308-00 BB, GE Healthcare) containing 8.3 ml of Sephadex™ G-25 resin.

Short-term transfection of CXCR family members into Expi293F™ cells

Expi293F™ cells (Life Technologies) were transiently transfected with the commercially available mammalian expression vector pTT5 (Durocher, 2002) containing a polynucleotide encoding: human CXCR1 (SEQ ID NO: 133); or human CXCR2 (SEQ ID NO: 125); or human CXCR3 (SEQ ID NO: 128); or human CXCR4 (SEQ ID NO: 129); or human CXCR5 (SEQ ID NO: 130); or human CXCR6 (SEQ ID NO: 131); or human CXCR7 (SEQ ID NO: 132); or cynomolgus CXCR2 (SEQ ID NO: 127).

For each 10 mL transfection, lipid-DNA complexes were prepared by diluting 10 μg of plasmid DNA in Opti-MEM™ I Reconstituted Serum Medium (Cat. No. 31985-062) to a total volume of 1.0 mL. Dilute 54 µl of ExpiFectamine™ 293 reagent in Opti-MEM™ I medium to a total volume of 1.0 ml. Transfection was performed according to the manufacturer's instructions. Cells were incubated at 37°C in a humidified incubator with 8% CO ₂ in air on a circular shaker at 200 rpm. Approximately 18-24 hours after transfection, cell viability was determined and transfected cells were harvested for use.

Flow cytometry binding assay on short-term transfected Expi293F™ cells

Collected cells were resuspended in FACS buffer (1×PBS + 0.5% (w/v) bovine serum albumin (BSA) + 2 mM EDTA, pH 7.2). Approximately 2×10 ⁵ cells per well were added to a 96-well round-bottom plate. Cells were pelleted by centrifugation at 400 g for 5 min at 4°C, and the supernatants were discarded. 25 μl of each test antibody or control antibody was added to the cells and incubated for 30 minutes at 4°C. Cells were washed twice with 100 μl of FACS buffer, followed by centrifugation at 400 g for 5 min at 4°C.

For detection, 50 μl of secondary antibody was added to the corresponding samples (Table 2). Commercially available antibodies binding to transfected CXCR family members were used as a positive control to ensure that the cells expressed the receptors. Cells were washed twice with 200 μl of FACS buffer, followed by centrifugation at 400 g for 5 min at 4°C. Before sample analysis, cells were resuspended in 100 μl FACS buffer. Samples were analyzed on a FACSCanto™ II cytometer (Beckton-Dickinson) using a high-throughput sampler.

Table 2. Reagents used for flow cytometry method

Reagent Catalog no. Manufacturer or supplier Recommended final dilution Anti-human CXCR1 antibody with FITC FAB330F R&D Systems 1 to 10 Anti-human CXCR2 antibody with FITC 551126 BD Biosciences 1 to 5 Anti-human CXCR3 antibody with FITC 558047 BD Biosciences 1 to 10 Anti-human CXCR4 antibody with FITC 561735 BD Biosciences 1 to 10 Anti-human CXCR5 antibody with FITC 558112 BD Biosciences 1 to 10 Human CXCR6 antibody with PE 356004 BioLegend 1 to 20 Human CXCR7 antibody with PE 331104 Bio Legend 1 to 20 Human IgG4, kappa (isotype control) I4639-1MG Sigma-Aldrich 10 µg/ml Anti-human IgG-Fc antibody with FITC F9512-2ML Sigma-Aldrich 1 to 200 Anti-human Ig λ light chain antibody with APC 316610 BioLegend 1 to 20 Antibody to human CXCR2 with APC 320710 BioLegend 1 to 20 Anti-human CXCR2 antibody with Alexa Fluor™ 647 FAB331R-100 R&D Systems 1 to 20 Antibody against human CXCR2 555932 BD Biosciences Antibody against human CXCR2 MAB331 R&D Systems 7AAD 559925 BD Biosciences 1 in 50

Analysis of activity on cells with human CXCR2

Tango™ cell assay with CXCR2. To assess the ability of antibodies to inhibit CXCR2 activation by CXCL8 and CXCL1, a commercially available Tango™ CXCR2-bla U2OS reporter cell line (ThermoFisher Scientific, Australia) was used. Cells were thawed, expanded, cultured, and frozen according to the manufacturer's instructions.

Preparation of Tango™ CXCR2-bla U2OS cells for use in cell-based assays. The manufacturer's protocol was modified to use 96-well plates instead of 384-well plates. Briefly, dividing cells were collected the day before use. Cells were harvested and resuspended in assay medium (100% FreeStyle™ Expression Medium; Life Technologies; cat. #12338-018) at a viable cell density of 312,500 cells/mL. 128 μl of cell suspension per well was added to 96-well plates with black walls and a transparent bottom, treated for tissue culture. Cells were incubated for 16-20 hours at 37°C in an atmosphere of 5% CO ₂ before being used in the assay.

Tango™ Analysis Procedure. Assays were set up and performed as described in the manufacturer's protocol. Agonists used in the analysis of both agonist and antagonist activity are presented in Table. 3. For antagonist assays, agonists were used at concentrations in the EC ₅₀ -EC ₈₀ range. Data were read on a FlexStation ^® 3 fluorescence reader (Molecular Devices) configured with the parameters shown in Table. 4. The blue/green emission ratios for each well were calculated by dividing the blue emission values by the green emission values. All inhibition curves were generated using four dose-response parameters using GraphPad Prism™ (version 7.01).

Table 3. Agonists used in cellular activity assays

Agonist for humans Manufacturer Catalog no. EC ₅₀ (nM) according to Tango™ EC ₅₀ (nM) for calcium fluxes average range average range CXCL1 Miltenyi Biotec 130-108-974 2.30 1.5- 3.4 2.62 0.7-6.7 CXCL2 R&D Systems 276-GB 33.3 15-45.7 37.02 not enough data CXCL3 R&D Systems 277-GG 11.9 6.0-22.7 12.25 10-18 CXCL5 R&D Systems 254-XB 97.7 47.7-150 39.17 10.3-20.5 CXCL6 R&D Systems 333-GC 9.40 5.60-12.4 28.94 26.8-32.4 CXCL7 R&D Systems 393-NP 39.6 15.3-77.2 7.14 5.3-9.0 CXCL8 Miltenyi Biotec 130-108-979 2.20 1.6-4.0 3.20 0.9-6.5

Table 4: FlexStation ^® 3 Fluorescence Reader Settings

Scan 1 (blue) Scan 2 (green) Excitation filter 409/20 nm 409/20 nm Radiation filter 460/40 nm 530/30 nm

Peripheral blood products

Whole human buffy coat, PBMC-enriched fractions obtained from human peripheral blood buffy coat, or whole blood of cynomolgus macaques were used for flow cytometry analysis of antibody binding. Briefly, blood was left untreated or diluted 1:1 in sterile room temperature filtered phosphate-buffered saline (PBS). 30 ml samples were layered onto 15 ml Lymphoprep™ (Stem Cell Technologies, cat. #07851). Peripheral blood mononuclear cells (PBMCs) were enriched by centrifugation at room temperature at 700 g for 30 min without inhibition. The PBMC layer was removed and the cells were washed with PBS containing 2 mM EDTA with low-speed centrifugation at 200 g to remove platelet contamination. Whole blood or PBMC-enriched fractions were resuspended in red blood cell lysis solution (BioLegend, 420301). The number of viable cells was determined and the cells were resuspended at 1×10 ⁷ cells per ml in FACS buffer (1×PBS + 0.5% (w/v) BSA + 2 mM EDTA, pH 7.2).

Flow cytometric binding assay on whole blood or PBMC-enriched preparations. Binding assays to detect CXCR2 in blood neutrophil populations using the test antibodies were essentially performed as described for the transiently transfected Expi293F™ cell assay. Antibody binding to human CXCR2 on the surface of neutrophils was measured using anti-CXCR2 antibodies directly conjugated to the fluorophore allophycocyanin (APC). Appropriate isotype control antibodies were included for comparison. Cells were incubated with lineage-specific antibodies and 2 μg/ml APC-conjugated anti-CXCR2 antibody or isotype control prepared in ice-cold 3% BSA/PBS for at least 30 min at 4°C. A commercial anti-CXCR2 antibody (clone 48331, R&D Systems FAB331A) was used as a positive control. Neutrophil populations were identified by their characteristic granularity and size, as well as by the binding of commercial anti-CD10 antibodies (BioLegend, clone Hl10a, cat. no. 312204). Cells were washed and fixed (BioLegend Fixation Buffer, 420801) before analysis. The level of fluorescence on the cell surface was measured by flow cytometry. This method showed that the tested anti-CXCR2 antibodies bind to both human and cynomolgus macaque neutrophils.

Generation of transgenic mice with human CXCR2

hCXCR2-introduced mice were generated by homologous recombination in embryonic stem (ES) cells with the insertion of human CXCR2 into exon 1 of the mouse CXCR2 gene. The single coding exon of human CXCR2 was inserted into a vector with 5′ and 3′ arms homologous to the genomic location of the mouse CXCR2 coding exon, yielding a gene structure encoding human CXCR2 but retaining mouse noncoding and regulatory elements. This vector was electroporated into ES cells from C57Bl/6 mice, which were introduced into blastocysts from C57Bl/6 mice and transplanted into pseudopregnant female mice. The pups were backcrossed to the original C57Bl/6 line, and the progeny were tested by Southern blotting for gametic transmission of the human CXCR2 gene. Mice heterozygous for human CXCR2 were crossed with each other to produce a line homozygous for human CXCR2, and the phenotype was tested by binding of antibodies against human and mouse CXCR2 by flow cytometry.

Comparative antibody and small molecule antagonists of CXCR2

Antagonist 1 is the HY29GL antibody described in Int. Pub. No. WO2015/169811 A2 (V _H and V _L of SEQ ID NO: 20 and 29 of this link). Antagonist 2 - antibody CX1_5, described in Int. Pub. No. WO2014/170317 A1 (V _H and V _L of SEQ ID NOs: 115 and 114 of this link). Antagonist 3 - anti-CXCR2 antibody clone 48311 (R&D Systems, cat. no. MAB331-500). Antagonist 4 - anti-CXCR2 antibody clone 6C6 (BD Biosciences, cat. #555932). Antagonist 5 is the small molecule CXCR2 antagonist danirixin. Antagonist 6 is a small molecule antagonist of CXCR2 SCH 527123.

Example 1: Preparation of anti-CXCR2 antibodies

Production of antibodies against CXCR2 with human variable regions

To generate antibodies against human CXCR2, transgenic rats engineered to express human variable region antibodies were used (as described in Int. Pub. No. WO2008/151081). Briefly, animals were genetically immunized once a week with a plasmid encoding the amino acid sequence of human CXCR2 (SEQ ID NO: 125) until anti-human CXCR2 antibody titers were obtained as measured by flow cytometry in CXCR2 positive transients. HEK-293 cells.

Production and propagation of hybridomas expressing antibodies against CXCR2

To obtain hybridomas that produce monoclonal antibodies to human CXCR2, splenocytes and/or lymph node cells from animals with the highest titers of anti-CXCR2 antibodies were isolated and fused with mouse myeloma cells (ATCC, CRL-1580). Cells were seeded at approximately 1×10 ⁵ cells/ml in flat-bottomed plates, followed by two weeks of incubation in selective media (10% FCS and 1×HAT (Sigma). Hybridomas were propagated by serial passage with four changes of media in 96-well plates (96- well stages 1-4) and then, if necessary, expanded into T25 and T75 flasks. During hybridoma propagation, supernatants were monitored for CXCR2 binding activity by cellular ELISA (cELISA) on cells transiently transfected to express human CXCR2 or mouse CXCR2 ( SEQ ID NO: 125 or 126, respectively) Bound antibodies were detected using secondary goat anti-rat IgG-HRP antibody (Southern Biotech, #3030-05).

Using lymph node and spleen cells from transgenic rats engineered to express human variable region sequences, panels of hybridomas were prepared. To detect binding activity to human CXCR2 in supernatants obtained during hybridoma propagation, the cell-based ELISA (cELISA) method was used (Table 5). Hybridomas that retained expression of CXCR2-binding antibodies after several passages were selected for DNA sequencing.

Table 5. Binding of hybridoma supernatants to cells transfected with human CXCR2 or mouse CXCR2 by cELISA

CLONE cELISA on rat hybridoma supernatants with human variable regions Human CXCR2 CXCR2 mice % positive* % positive* BKO-1A1 38% 6% BKO-1B10 75% 4% BKO-1C1 3% 5% BKO-1C6 88% 5% BKO-1D1 67% 7% BKO-1D5 46% 6% BKO-1D9 85% 5% BKO-1E3 3% 6% BKO-1H3 41% 3% BKO-2A3 3% 3% BKO-2B10 79% 5% BKO-2C2 77% 5% BKO-2D1 28% 4% BKO-2D8 100% 7% BKO-2E4 80% 7% BKO-2F6 eleven% 3% BKO-2G4 83% 4% BKO-2G7 75% 3% BKO-2G10 139% 4% BKO-2H4 2% 3% BKO-2H7 23% 5% BKO-3A9 2% 4% BKO-3C3 8% 10% BKO-3D3 3% 6% BKO-3D6 2% 3% BKO-3E3 107% 3% BKO-3F4 86% 42% BKO-3F5 5% 5% BKO-3F6 6% 5% BKO-3G11 61% 7% BKO-3H11 24% 5% BKO-4A4 32% 7% BKO-4A5 2% 7% BKO-4A8 110% 5% BKO-4A10 eleven% 5% BKO-4B2 107% 6% BKO-4B7 2% 3% BKO-4B11 103% 7% BKO-4C1 97% 6% BKO-4E8 53% 7% BKO-4F10 118% 7% BKO-4F11 77% 8% BKO-4G3 2% 2% BKO-4G4 93% 3% BKO-4H5 81% 4% BKO-4H6 108% 4% BKO-4H11 4% 6% BKO-5A5 66% 5% BKO-5B8 53% 5% BKO-5C4 101% 5% BKO-5E8 109% 8% BKO-5F9 2% 4% BKO-5F10 119% 4% BKO-5G6 103% 4% BKO-5G11 101% 22% BKO-5H1 4% 7% BKO-5H4 5% 8% BKO-6A1 60% 6% BKO-6A2 4% 7% BKO-6A3 8% 7% BKO-6B8 53% 4% BKO-6C2 3% 6% BKO-6C4 39% 5% BKO-6D10 2% 4% BKO-6E4 79% 3% BKO-6F3 44% 6% BKO-6G1 101% 4% BKO-6H1 52% 5% BKO-7A3 21% 8% BKO-7A9 2% 4% BKO-7B1 15% 5% BKO-7B2 84% 4% BKO-7C11 73% 4% BKO-7D8 67% 6% BKO-7D9 5% 7% BKO-7E4 4% 6% BKO-7E7 94% 12% BKO-7F3 95% 7% BKO-7F11 2% 3% BKO-7G1 21% 3% BKO-7G2 2% 8% BKO-7G10 104% 26% BKO-7H7 3% 7% BKO-7H8 105% 7% BKO-7H11 4% 9% BKO-8B6 95% 6% BKO-8C2 5% 8% BKO-8C4 124% 6% BKO-8D2 eleven% 7% BKO-8D4 38% 7% BKO-8F3 56% 3% BKO-8G3 114% 6% BKO-8H8 104% 7% BKO-8H10 81% 8% BKO-9A8 124% 8% BKO-9C3 56% 7% BKO-9C4 2% 3% BKO-9E7 2% 3% BKO-9G1 66% 4% BKO-9G6 34% 8% BKO-9H5 4% 7% BKO-10A2 25% 5% BKO-10B4 109% 6% BKO-10D1 2% 6% BKO-10D8 thirty% 10% BKO-10F3 7% 4% BKO-10G10 89% 4% BKO-10H6 6% 6% Positive control 100% 100% Negative control 4% 8%

* Fluorescence units relative to positive control (100%)

Sequencing of antibodies produced by hybridoma cells

Variable domains of antibodies were isolated by reverse transcription-polymerase chain reaction (RT-PCR), using RNA obtained from sediments of nonclonal hybridoma cells as a template. RNA was isolated from hybridoma plates using the Genelute™ 96-well Total RNA Purification Kit (Sigma #RTN9602, RTN9604) according to the manufacturer's procedure. For standard RT-PCR, RNA was reverse transcribed into cDNA using the oligo(dT) primer and the AccuScript PfuUltra ^® II RT-PCR kit (Agilent #600184). The cDNA synthesis reactions were prepared according to the manufacturer's procedure, and cDNA synthesis was carried out at 42°C for 30 min. For PCR with rapid 5′ amplification of cDNA ends (5′-RACE), reverse transcription of RNA into cDNA was performed using the SMARTer ^® RACE kit (Takara) according to the manufacturer's instructions, resulting in cDNA ready for 5′-RACE.

Amplification of human antibody variable regions from a panel of hybridomas was performed by PCR using PfuUltra II (Agilent) or Q5 (NEB) high-fidelity DNA polymerase according to the manufacturer's instructions. Heavy chains from the hybridoma kit were amplified using primer pairs specific for the rodent heavy chain constant region DNA sequence and human heavy chain leader DNA sequences. Lambda light chain variable regions from the hybridoma panel were amplified in the same manner using primer pairs specific for the human lambda constant region DNA sequence and human lambda chain leader DNA sequences.

The concentration of the resulting purified DNA was determined using a spectrophotometer using the Nanodrop method. Sanger sequencing of PCR fragments was performed using oligonucleotides designed to bind within heavy or light chain amplicons. The resulting DNA sequences were subjected to virtual translation into amino acid sequences for further analysis before their use in the production of full-length antibody chains. Antibodies with unique amino acid sequences were selected for conversion to full-length human antibodies.

Recombinant monoclonal antibodies binding to human CXCR2

Hybridomas selected from those that secrete antibodies that bind to CXCR2 (as described in Table 5) were sequenced to identify variable region DNA and amino acids by RT-PCR as described above. These antibody variable regions were then prepared by gene synthesis and subcloned into mammalian expression vectors as described in General Methods. Antibodies were generated by coexpression of heavy and light chain plasmids in Expi293F™ cells and purified by protein A column chromatography as described in General Methods. When multiple heavy and/or light chains were identified from the same hybridoma cells, each heavy chain was paired with each light chain, and the resulting antibodies were given the suffixes a, b, c, etc. Purified antibodies were desalted in Sorensen's PBS pH 5.8 and tested by flow cytometry for binding to Expi293F™ cells transfected with human CXCR2 or human CXCR1, as well as to mock-transfected Expi293F™ cells. From the hybridoma set, 26 antibodies were identified for further characterization that bind to human CXCR2, but not to human CXCR1 or to mock-transfected Expi293F™ cells. The sequences of these antibodies are given in table. 6.

Table 6. Sequences of human variable region antibodies binding to human CXCR2 but not to the closely related human CXCR1 or to mock-transfected Expi293F™ cells

Antibody name V _H (SEQ ID NO) V _L (SEQ ID NO) BKO-1A1 BKO_1A1_VH
(SEQ ID NO: 1) BKO_1A1_VL
(SEQ ID NO: 2) BKO-1B10 BKO_1B10_VH
(SEQ ID NO: 3) BKO_1B10_VL
(SEQ ID NO: 4) BKO-1D1 BKO_1D1_VH
(SEQ ID NO: 5) BKO_1D1_VL
(SEQ ID NO: 6) BKO-1H3 BKO_1H3_VH
(SEQ ID NO: 7) BKO_1H3_VL
(SEQ ID NO: 8) BKO-2D8 BKO_2D8_VH
(SEQ ID NO: 9) BKO_2D8_VL
(SEQ ID NO: 10) BKO-3A9_b BKO_3A9_VH
(SEQ ID NO: 11) BKO_3A9_L3_E03_VL
(SEQ ID NO: 12) BKO-3D6 BKO_3D6_VH
(SEQ ID NO: 13) BKO_3D6_L6_G06_VL (BKO_5H4_VL)
(SEQ ID NO: 14) BKO-3F4 BKO_3F4_VH
(SEQ ID NO: 15) BKO_3F4_L11_A11_VL
(SEQ ID NO: 16) BKO-4A8 BKO_4A8_VH
(SEQ ID NO: 17) BKO_4A8_VL
(SEQ ID NO: 18) BKO-4F10 BKO_4F10_VH
(SEQ ID NO: 19) BKO_4F10_VL
(SEQ ID NO: 20) BKO-5E8 BKO_5E8_H5_C05_VH
SEQ ID NO: 21) BKO_5E8_L3_C03_VL
(SEQ ID NO: 22) BKO-5G11 BKO_5G11_VH
(SEQ ID NO: 23) BKO_5G11_VL
(SEQ ID NO: 24) BKO-5G6_c BKO_5G6_VH
(SEQ ID NO: 25) BKO_5G6_L12_E12_VL
(SEQ ID NO: 26) BKO-6A1_b BKO_6A1_H4_A04_VH
(SEQ ID NO: 27) BKO_6A1_E10_VL
(SEQ ID NO: 28) BKO-6A2_a BKO_6A2_H1_B01_VH
(SEQ ID NO: 29) BKO_6A2_L6_A06_VL
(SEQ ID NO: 30) BKO-7C11 BKO_7C11_H6_B06_VH
(SEQ ID NO: 31) BKO_7C11_G01_VL
(SEQ ID NO: 32) BKO-7G10_a BKO_7G10_H1_B01_VH
(SEQ ID NO: 33) BKO_7G10_L6_E06_VL
(SEQ ID NO: 34) BKO-7H8_b BKO_7H8_H3_C03_VH
(SEQ ID NO: 35) BKO_7H8_L10_F10_VL
(SEQ ID NO: 36) BKO-8B6 BKO_8B6_VH
(SEQ ID NO: 37) BKO_8B6_VL
(SEQ ID NO: 38) BKO-8C4 BKO_8C4_VH
(SEQ ID NO: 39) BKO_8C4_VL
(SEQ ID NO: 40) BKO-8G3_b BKO_8G3_H4_D04_VH
(SEQ ID NO: 41) BKO_8G3_L1_G01_VL
(SEQ ID NO: 42) BKO-8H10 BKO_8H10_VH
(SEQ ID NO: 43) BKO_8H10_VL
(SEQ ID NO: 44) BKO-8H8_b BKO_8H8_H5_E05_VH
(SEQ ID NO: 45) BKO_8H8_L7_H08_VL
(SEQ ID NO: 46) BKO-9A8 BKO_9A8_H3_F03_VH
(SEQ ID NO: 47) BKO_9A8_L1_H02_VL
(SEQ ID NO: 48) BKO-9C3_a BKO_9C3_H8_G08_VH
(SEQ ID NO: 49) BKO_9C3_L1_F01_VL
(SEQ ID NO: 50) BKO-10G10 BKO_10G10_VH
(SEQ ID NO: 51) BKO_10G10_VL
(SEQ ID NO: 52)

Example 2. Functional characterization of anti-CXCR2 antibodies

Binding to cynomolgus CXCR2

All recombinant antibodies that bind to human CXCR2, but not to human CXCR1 or to mock-transfected Expi293™ cells, were tested for binding to cynomolgus CXCR2 (SEQ ID NO: 127) on Expi293F™ cells transiently transfected with a plasmid encoding the CXCR2 protein cynomolgus macaque, as described here. Antibodies with detectable levels of binding to cynomolgus CXCR2 were further studied.

Association with other members of the human CXCR family

Antibodies cross-reactive with human and cynomolgus CXCR2 were tested by flow cytometry for binding to other human CXCR family members CXCR3, CXCR4, CXCR5, CXCR6 and CXCR7 on Expi293F™ cells transiently transfected with a plasmid encoding CXCR3 (SEQ ID NO: 128), CXCR4 (SEQ ID NO: 129), CXCR5 (SEQ ID NO: 130), CXCR6 (SEQ ID NO: 131) or human CXCR7 (SEQ ID NO: 132). Antibodies that were not selective for CXCR2 were excluded from further analysis.

Inhibition of CXCL8-mediated activation of human CXCR2

Antibodies were tested on Tango™ cells with CXCR2 using CXCL8 as the agonist as described in General Methods. As can be seen from Fig. 1A and 1B, eight antibodies inhibited CXCL8-induced CXCR2 activation. In this case, the commercially available anti-CXCR2 antibody 6C6 (BD Biosciences; “BD6C6”) was used as a positive control.

Antibody binding to human CXCR2 expressed in human PBMC and to cynomolgus CXCR2 expressed in cynomolgus PBMC

Eight antibodies that inhibited CXCL8-mediated activation of CXCR 2 were tested for binding to native CXCR2 expressed in human and cynomolgus macaque PBMCs. As can be seen from Fig. 2, two antibodies, BKO-4A8 and BKO-8G3_b, exhibited high binding activity to human and cynomolgus CXCR2, while the third, BKO-9C3_a, exhibited significant levels of binding to human CXCR2 and above average binding to cynomolgus CXCR2.

Inhibition of CXCL1-mediated activation of human CXCR2

Antibodies BKO-4A8, BKO-8G3_b and BKO-9C3_a were tested on Tango™ cells with CXCR2 using CXCL1 as the agonist as described in General Methods. The BKO-4A8 antibody was the strongest of the antibodies tested in this format. Typical CXCL1 inhibition of CXCR2 activation curves for these antibodies are shown below in FIG. 3.

Example 3: Optimization of the amino acid sequence of the anti-CXCR2 antibody BKO-4A8

Heavy and light chain variable region variants of the anti-CXCR2 antibody BKO-4A8 were designed to optimize the sequence of this molecule to ensure stability and manufacturability.

Variants of the BKO-4A8 antibody containing one amino acid substitution were generated as described below. A summary of these options is presented in Table. 7.

Table 7. BKO-4A8 variants

Antibody chain Replacement SEQ ID NO: variable region Heavy, CDR1 S32Q 53 S32D 136 S32H 54 S32L 55 S32W 56 S32Y 57 T33A 58 M34Q 59 M34D 60 M34H 61 M34W 62 S35Q 137 S35D 138 S35K 139 Heavy, CDR2 A50S 140 I51H 63 G52aD 64 R53S 65 R53Q 66 G54D 67 R55Q 141 R55D 142 R55H 143 N56S 68 Heavy, CDR3 I94K 69 M96A 70 M96S 144 M96Q 71 M96D 145 M96H 146 M96K 72 M96L 147 M96W 148 M96Y 149 G101D 73 Y102S 74 Y102Q 150 Y102D 151 Y102K 75 Lightweight, CDR2 E50D 76 V51D 152 V51Y 153 N52D 77 N52S 78 K53A 79 K53D 80 K53H 81 R54Q 82 R54D 154 Lightweight, CDR3 Y91A 83 Y91S 155 Y91H 156 N94A 84 N94S 85 N94H 157 N94K 86 N94L 87 N94W 88 N94Y 89 N95aS 158 N95aQ 90 N95aD 91 N95aH 92 N95aK 93 N95aL 94 N95aW 159 N95aY 95 V97A 96 V97S 160 V97D 161 V97K 97

Preparation of plasmids encoding antibody variants using site-directed mutagenesis

Plasmids encoding antibody chains requiring substitution at a single amino acid were obtained by replicating plasmid strands under the action of mutagenic primers using high-precision DNA polymerase. This process uses supercoiled double-stranded plasmid DNA as a template and two complementary synthetic oligonucleotide primers, both containing the desired mutation. Oligonucleotide primers complementary to each of the opposite strands of the plasmid undergo elongation in cyclic PCR reactions without primer displacement, thereby producing copies of the mutant plasmid containing stepwise cuts. After a round of PCR reactions, they were treated with the restriction enzyme DpnI. DpnI preferentially cuts the original vector DNA, leaving the newly synthesized strands intact.

To generate variants involving multiple amino acid substitutions that required mutagenic oligonucleotide primers longer than 40 nucleotides, the above process was repeated to introduce substitutions sequentially, or genes were synthesized de novo by assembling synthetic oligonucleotides. The sequences of all DNA constructs were verified by Sanger DNA sequencing before use.

Obtaining plasmids encoding antibody variants by polynucleotide synthesis

The amino acid sequences of the variable regions were reverse translated into DNA sequences using GeneOptimizer ^® technology before synthesis of the resulting de novo DNA by assembling synthetic oligonucleotides (GeneArt, Germany). Polynucleotides encoding synthesized heavy chain variable regions coupled with a human IgG4 constant region containing the S228P hinge stabilizing substitution (SEQ ID NO: 118) were subcloned into an expression vector. Polynucleotides encoding the synthesized lambda light chain variable region genes were subcloned into an expression vector encoding the human lambda light chain constant region (SEQ ID NO: 134).

Linking BKO-4A8 variants to CXCR2

Each heavy chain variant was expressed together with the original light chain and vice versa. The resulting antibodies were purified and tested for binding to CXCR2 relative to the parent antibody BKO-4A8 as described here. Antibody variants with binding levels close to the original BKO-4A8 are presented in Table. 8.

Table 8. BKO-4A8 point variants with levels of CXCR2 binding close to the original BKO-4A8

Antibody chain Replacement SEQ ID NO: variable region Binding level relative to BKO-4A8 Heavy, CDR1 S32Q 53 1.71 S32H 54 0.86 S32L 55 1.33 S32W 56 0.83 S32Y 57 1.33 T33A 58 1.30 M34Q 59 1.00 M34D 60 1.60 M34H 61 0.80 M34W 62 1.17 Heavy, CDR2 I51H 63 1.80 G52aD 64 1.18 R53S 65 1.00 R53Q 66 0.64 G54D 67 1.00 N56S 68 1.00 Heavy, CDR3 I94K ^# 69 0.7 M96A 70 1.5 M96Q 71 1.5 M96K 72 1.0 G101D 73 1.0 Y102S 74 1.5 Y102K 75 1.3 Lightweight, CDR2 E50D 76 2.0 N52D 77 1.7 N52S 78 1.0 K53A 79 1.4 K53D 80 1.4 K53H 81 1.4 R54Q 82 1.6 Lightweight, CDR3 Y91A 83 1.7 N94A 84 1.0 N94S 85 1.0 N94K 86 1.33 N94L 87 0.88 N94W 88 0.63 N94Y 89 0.50 N95aQ 90 1.0 N95aD 91 2.3 N95aH 92 1.3 N95aK 93 1.0 N95aL 94 1.7 N95aY 95 1.7 V97A 96 1.7 V97K 97 1.0

# Indicates that this framework residue flanks the CDR3 of the heavy chain

In fig. 4A and 4B are sequence alignments of the heavy chain variable region and light chain variable region, respectively, of the above antibody variants. Also shown in these figures is a heavy chain consensus variable region (“consensus V _H ”; SEQ ID NO: 167) and a light chain consensus variable region (“consensus V _L ”; SEQ ID NO: 168).

BKO-4A8 point variants with similar efficacy to BKO-4A8 in inhibiting CXCL1 or CXCL8-induced CXCR2 activation

Sixteen antibody variants exhibiting similar levels of binding to CXCR2 as the parent BKO-4A8 were tested for efficacy using CXCL1 or CXCL8 as the agonist as described in General Methods. As can be seen from table. 9, twelve antibodies exhibited potencies similar to the parent BKO-4A8. In fig. 4A and 4B show the amino acid sequence alignment of these variants relative to the parent heavy and light chain sequences.

Table 9. BKO-4A8 point variants with similar efficacy to BKO-4A8 in inhibiting CXCL1 or CXCL8-induced CXCR2 activation

Antibody chain Replacement SEQ ID NO: variable region IC ₅₀ on Tango cells (nM) CXCL1 CXCL8 BKO-4A8 - 17 0.177 0.112 Heavy, CDR1 M34Q 59 0.242 0.0776 M34H 61 0.186 0.107 Heavy, CDR2 N56S 68 0.158 0.129 Heavy, CDR3 I94K ^# 69 0.078 0.071 M96A 70 0.140 0.126 M96Q 71 0.330 0.140 M96K 72 0.135 0.091 Lightweight, CDR2 N52S 78 0.214 0.126 Lightweight, CDR3 N94S 85 0.149 0.169 N94K 86 0.123 0.113 N94Y 89 0.164 0.105 N95aQ 90 0.213 0.084

# Indicates that this framework residue flanks the CDR3 of the heavy chain

Combinatorial variants of BKO-4A8

The 3 non-germline framework amino acids were replaced back to those in the closest human germline sequence R75K and I94K in the heavy chain and D41G in the light chain. An A40P substitution was also introduced into framework region 2 of the heavy chain.

A panel of combinatorial variants containing two or more amino acid substitutions was compiled to explore the possibility of further optimization of these sequences. In table 10 shows all two light chain variants and 11 heavy chain variants that were obtained. In fig. 5A and 5B show amino acid sequence alignments with the heavy chain variable region and light chain variable region of the parent BKO-4A8, respectively. Also shown in these figures is a heavy chain consensus variable region (“consensus V _H ”; SEQ ID NO: 226) and a light chain consensus variable region (“consensus V _L ”; SEQ ID NO: 227).

Table 10. Combinatorial variants of BKO-4A8

Chain Option Substitutions (per BKO-4A8: relative to the original HC variable region of SEQ ID NO: 17 or the original LC variable region of SEQ ID NO: 18) Heavy 1 M34Q, N56S (SEQ ID NO: 98) 2 M34Q, A40P, N56S (SEQ ID NO: 99) 3 M34Q, A40P, N56S, R75K (SEQ ID NO: 100) 4 M34Q, A40P, N56S, M96K (SEQ ID NO: 101) 5 M34Q, A40P, N56S, R75K, M96K (SEQ ID NO: 102) 6 M34Q, A40P, N56S, R75K, I94K, M96K (SEQ ID NO: 103) 7 M34Q, A40P, N56S, I94K, M96K (SEQ ID NO: 104) 8 M34Q, N56S, M96K (SEQ ID NO: 105) 9 M34Q, N56S, R75K, M96K (SEQ ID NO: 106) 10 I94K, M96K (SEQ ID NO: 107) 101 M34Q, A40P, N56S, R75K, M96A (SEQ ID NO: 108) Lightweight b N52S, N94S (SEQ ID NO: 109) c D41G, N52S, N94S (SEQ ID NO: 110)

Each heavy chain was expressed together with each light chain to produce sequence optimized antibody variants. Antibodies were purified and tested for CXCR2 binding activity as described in General Methods. As can be seen from table. 11, most combinatorial antibody variants retained CXCR2 binding activity similar to the parent antibody BKO-4A8.

Table 11. Summary of EC ₅₀ values for linking combinatorial variants to CXCR2

Antibody name Substitutions in V _H Substitutions in V _L EC ₅₀ (nM) for binding c
CXCR2 Improvement (times) before
4A8 wild type 1b M34Q, N56S N52S, N94S
(SEQ ID NO: 109) 0.3 1.33 1c (SEQ ID NO: 98)
D41G, N52S, N94S (SEQ ID NO: 110) 0.3 1.33 Control 4A8 for V _H 1 n/a n/a 0.4 n/a 2b M34Q, A40P, N56S N52S, N94S
(SEQ ID NO: 109) 0.4 1.25 2c (SEQ ID NO: 99)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.3 1.67 Control 4A8 for V _H 2 n/a n/a 0.5 n/a 3b M34Q, A40P, N56S, R75K N52S, N94S
(SEQ ID NO: 109) 0.1 4.00 3c (SEQ ID NO: 100)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.2 2.00 Control
4A8 for _VH 3 n/a n/a 0.4 n/a 4b M34Q, A40P, N56S, M96K N52S, N94S
(SEQ ID NO: 109) 0.4 1.25 4c (SEQ ID NO: 101)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.4 1.25 Control
4A8 for _VH 4 n/a n/a 0.5 n/a 5b M34Q, A40P, N56S, R75K, M96K N52S, N94S
(SEQ ID NO: 109) 0.4 1.25 5c (SEQ ID NO: 102)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.4 1.25 Control
4A8 for _VH 5 n/a n/a 0.5 n/a 6b M34Q, A40P, N56S, R75K, I94K, M96K N52S, N94S
(SEQ ID NO: 109) 0.4 1.00 6c (SEQ ID NO: 103)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.9 0.44 Control 4A8 for V _H 6 n/a n/a 0.4 n/a 7b M34Q, A40P, N56S, I94K, M96K N52S, N94S
(SEQ ID NO: 109) 0.8 0.63 7c (SEQ ID NO: 104)
D41G, N52S, N94S
(SEQ ID NO: 110) 1.0 0.50 Control
4A8 for _VH 7 n/a n/a 0.5 n/a 8b
M34Q, N56S, M96K N52S, N94S
(SEQ ID NO: 109) n/a n/a 8c (SEQ ID NO: 105)
D41G, N52S, N94S
(SEQ ID NO: 110) n/a n/a Control
4A8 for _VH 8 n/a n/a 0.6 n/a 9b M34Q, N56S, R75K, M96K N52S, N94S
(SEQ ID NO: 109) 0.3 3.00

9c (SEQ ID NO: 106)
D41G, N52S, N94S (SEQ ID NO: 110) 6.4 0.14 Control 4A8 for V _H 9 n/a n/a 0.9 n/a

10b I94K, M96K N52S, N94S
(SEQ ID NO: 109) 0.3 1.67 10c (SEQ ID NO: 107)
D41G, N52S, N94S
(SEQ ID NO: 110) 0.5 1.00 Control 4A8 for V _H 10 n/a n/a 0.5 n/a

Six more combinatorial variants of the heavy chain were obtained containing 4 or more amino acid substitutions, shown in Table. 12. An amino acid sequence alignment with the parent heavy chain of BKO-4A8 is shown in FIG. 5A. To generate antibody variants, each heavy chain was co-expressed with light chain c (SEQ ID NO: 110). Antibodies were purified and tested for CXCR2 binding activity as described in General Methods. CXCR2 binding activity was reduced in antibody 102c and antibody 106c.

Table 12. Summary of EC ₅₀ values for linking combinatorial variants to CXCR2

Antibody name Substitutions in V _H EC ₅₀ (nM) for binding c
CXCR2 Max. binding (MFI) Max. binding (MFI as % of 4A8 wt) 101c M34Q, A40P, N56S, R75K, M96A (SEQ ID NO: 108) 3.05 55048 89% 102c M34Q, A40P, N56S, R75K, M96E (SEQ ID NO: 162) 8.70 32980 53% 103c M34Q, A40P, R53S, R55G, N56S, R75K (SEQ ID NO: 163) 5.15 44445 72% Control 4A8
for 101c-103c n/a 1.58 62074 n/a 104c M34Q, A40P, R53S, R55G, N56S, R75K, M96K (SEQ ID NO: 164) 6.45 45097 54% 105c M34Q, A40P, R53S, R55G, N56S, R75K, M96A (SEQ ID NO: 165) 5.73 42741 52% 106c M34Q, A40P, R53S, R55G, N56S, R75K, M96E (SEQ ID NO: 166) > 100 11633 n/a Control 4A8 for 104c-106c n/a 3.82 82797 n/a

Antibodies were tested for potency using CXCL1 or CXCL8 as the agonist as described in General Methods. The combinatorial variants of antibodies 103c, 104c and 105c showed less efficiency compared to the parent antibody, while the 101c variant showed better efficiency compared to the parent antibody, as can be seen in Table. 13 and fig. 6A and 6B.

Table 13. Summary of IC ₅₀ values for the effectiveness of combinatorial options on CXCR2

Antibody name Substitutions in V _H
(SEQ ID NO: for _VH ) IC ₅₀ (nM)
by CXCL8 IC ₅₀ (nM)
by CXCL1 Improvement (times) on CXCL8 before 4A8 wt Improvement (times) on CXCL1 before 4A8 wt 101c M34Q, A40P, N56S, R75K, M96A (SEQ ID NO: 108) 0.28 0.63 2.39 1.49 103c M34Q, A40P, R53S, R55G, N56S, R75K (SEQ ID NO: 163) 1.25 2.40 0.53 2.55 104c M34Q, A40P, R53S, R55G, N56S, R75K, M96K (SEQ ID NO: 164) 0.75 2.17 0.88 0.43 105c M34Q, A40P, R53S, R55G, N56S, R75K, M96A (SEQ ID NO: 165) 1.85 4.00 0.36 0.24 Control 4A8 n/a 0.66 94.0 n/a n/a

Antibody 101c, containing the heavy chain variable region “101” and the light chain variable region “c”, was selected for cell-based efficacy evaluation because of its favorable properties observed. Antibody 101c was renamed BKO-4A8-101c. BKO-4A8-101c contains five optimizing substitutions in the heavy chain and three in the light chain. Comparison of BKO-4A8 with BKO-4A8-101c in a cell-based potency assay for their ability to inhibit CXCL1- or CXCL8-induced CXCR2 activation showed that sequence changes to optimize BKO-4A8-101c also increased its potency compared to the parent BKO-4A8. as can be seen from Fig. 6A and 6B.

Example 5 Characterization of CXCR2 Antagonism

Characterization of BKO-4A8-101c inhibition of ligand-mediated β-arrestin recruitment

Human CXCR2 Tango™ cells were used to evaluate the ability of the BKO-4A8-101c antibody to inhibit β-arrestin recruitment to agonist-activated CXCR2. All CXCR2 ligands of the human ELR+CXC chemokine type were tested in antagonist dose-response experiments using calculated EC values of ₅₀ agonists. BKO-4A8-101c inhibited CXCR2-mediated β-arrestin signaling induced by all ELR+CXC chemokines, with comparable IC ₅₀ values for all agonists tested (Table 14). BKO-4A8-101c completely inhibited human CXCR2 activation by human CXCL1, 2, 3, 5, and 6 in a dose-dependent manner, but only partially inhibited CXCL7 and CXCL8 in the same dose range. Representative data from 4 independent experiments are shown in Fig. 7.

Without committing to any proposed mechanism of action, it is believed that the observed selective antagonistic effect provides a therapeutic window allowing virtually complete inhibition of CXCL1- and CXCL5-mediated neutrophil migration from the circulation into tissues without significantly affecting CXCL8-mediated neutrophil migration from the bone marrow into the circulation. The partial inhibition of CXCL8-mediated β-arrestin recruitment in the reporter assay suggests that the β-arrestin-mediated receptor internalization pathway is functional.

Table 14. Antagonistic effect of BKO-4A8-101c on CXCR2 in a reporter assay for ligand-mediated β-arrestin recruitment (n = 7-13)

Human (hu) ligand IC ₅₀ (nM) Max. inhibition (%) average range average value Hu CXCL1 0.27 0.08-0.42 98 Hu CXCL2 0.36 0.23-0.44 100 Hu CXCL3 0.39 0.30-0.46 96 Hu CXCL5 0.30 0.11-0.47 98 Hu CXCL6 0.28 0.16-0.38 98 Hu CXCL7 0.44 0.29-0.62 76 Hu CXCL8 0.34 0.18-0.66 78

Characterization of BKO-4A8-101c inhibition of ligand-mediated calcium influx

One of the signaling pathways downstream of CXCR2 activation, which plays a role in cell chemotaxis, is characterized by calcium mobilization (influx). The ability of BKO-4A8-101c to inhibit human CXCR2 ligand-induced calcium influx was tested in the commercially available calcium-optimized human CXCR2 chemokine receptor cell line HTS002C-Chemiscreen™. All CXCR2 ligands of the human ELR+CXC chemokine type were tested in antagonist dose-response experiments using calculated EC values of ₅₀ agonists.

BKO-4A8-101c strongly inhibited human CXCL1, 2, 3, 5, and 6-induced calcium influx in a dose-dependent manner, but only weakly inhibited CXCL7 and modestly inhibited CXCL8 in the same dose range, as shown in Table 1. 15. Representative data for CXCL1, CXCL5 and CXCL8 are shown (Fig. 8).

The chemotactic response of neutrophils is known to be mediated through CXCR2-activated calcium mobilization. Without committing to any proposed mechanism of action, the selective antagonistic action of the antibodies described here potentially provides a therapeutic window allowing virtually complete inhibition of CXCL1- and CXCL5-mediated neutrophil migration into the lung without significantly affecting CXCL8-mediated neutrophil migration from the bone marrow into the bloodstream. This may help block neutrophil-mediated pathology at sites of chronic inflammation without necessarily compromising the original neutrophil-mediated antimicrobial functions.

Table 15. Summary of mean BKO-4A8-101c IC ₅₀ values for effects of human ELR+CXC chemokines on human CXCR2 in calcium influx assay (n = 4-9)

Human (hu) ligand IC ₅₀ (nM) Max. inhibition (%) average range average value Hu CXCL1 1.55 0.84 – 2.31 91 Hu CXCL2 1.62 0.75 – 2.30 88 Hu CXCL3 0.63 0.25 – 1.15 52 Hu CXCL5 1.03 n/ ^a 81 Hu CXCL6 1.71 1.57 – 2.00 81 Hu CXCL7 2.14 0.47 – 5.50 43 Hu CXCL8 n/o ^b - 9

^a Insufficient data ^b Does not inhibit or data are not suitable for constructing a four-point dose-response curve

Example 6. Binding and functional activity of anti-CXCR2 antibodies in vitro is independent of the Fc region

The heavy chain variable regions from BKO-4A8 were formatted onto the other human IgG constant regions shown in Table 1. 16. The ability of purified antibodies to bind to human CXCR2 was routinely assessed in Expi293F™ cells transiently transfected to express human CXCR2 (SEQ ID NO: 125). Binding of BKO-4A8 and its variants was detected by incubation of a fluorochrome-conjugated anti-human IgG lambda light chain antibody. The binding activity of the tested antibodies, measured as mean fluorescence intensity, was independent of the Fc region of the antibody, as can be seen in FIG. 9.

Table 16. BKO-4A8 heavy chain variants

Heavy chain variant BKO-4A8 SEQ ID NO: heavy chain BKO-4A8 IgG4* 115 BKO-4A8 IgG4 117 BKO-4A8 IgG2* 119 BKO-4A8 IgG1* 121 BKO-4A8 IgG1 123 * Means modified Fc

To assess the ability of anti-CXCR2 antibodies to inhibit β-arrestin recruitment to agonist-activated CXCR2, the commercially available Tango™ CXCR2-bla U2OS reporter cell line (ThermoFisher Scientific) was used. Agonists were added to samples at a concentration of EC ₅₀ for analysis of antagonists. The dose-response curves indicate that the functional activity of BKO-4A8 was not affected by sequence modification in the Fc region, with all antibodies tested exhibiting comparable antagonistic activity, as seen in FIG. 10.

Example 7 Anti-CXCR2 Antibody BKO-4A8-101c Specificity

The specificity of the anti-CXCR2 antibody BKO-4A8-101c was tested by assessing binding activity on Expi293F™ cells transiently transfected to express closely related human CXCR family members as shown in FIG. 11. The sequences of the studied representatives of the human CXCR family are given in table. 19 BKO-4A8 binding was detected by incubation of a fluorochrome-conjugated anti-human IgG antibody. The anti-CXCR2 antibody BKO-4A8-101c has been shown to bind strongly and selectively to human CXCR2.

Example 8 Flow Cytometry Binding Analysis on Primary Blood Cells

To further characterize the anti-CXCR2 antibody BKO-4A8-101c, its ability to bind to CXCR2 on neutrophils in anticoagulated human blood was tested. Binding was measured using BKO-4A8-101c conjugated directly to the APC fluorophore. For comparison, isotype-matched control antibodies conjugated to APC were included. Cells were incubated with lineage-specific antibodies and 2 μg/ml APC-conjugated anti-CXCR2 antibody or isotype control. The level of fluorescence on the cell surface was measured by flow cytometry. Cell debris and nonviable cells were excluded based on light scattering characteristics and inclusion of the viability fixable dye Zombie Violet (BioLegend, 423113). Hematopoietic cell subsets were identified based on CD45 expression along with characteristic size (forward scatter, FSC) and granularity (side scatter, SSC) together with the expression of phenotypic markers as follows: T lymphocytes = CD3; B lymphocytes = CD20; monocytes = CD14; and natural killer lymphocytes = CD56 (or CD3 ⁻ CD20 ⁻ CD16 ⁺ lymphocytes). Granulocytes were identified by size and granularity, as well as by the lack of binding to lineage-specific markers: CD3, CD19, CD20, CD56 and CD14. In addition, neutrophils were identified by high expression levels of CD16 and CD177, and eosinophils were identified by Siglec-8 expression. By this technique, BKO-4A8-101c binds to neutrophils (Fig. 12A) and monocytes (Fig. 12B) from human blood. A commercial anti-CXCR2 antibody (BioLegend, 5E8/CXCR2) was used as a positive control in the experiment.

Example 9: Ability of Purified Antibodies to Inhibit Agonist-Induced CXCR2-Mediated Functional Responses Compared to Comparative Antibodies and Small Molecules

Characterization of BKO-4A8-101c inhibition of ligand-mediated β-arrestin recruitment compared to comparative reagents

The ability of BKO-4A8-101c to block ligand-activated CXCR2 signaling was compared with comparative antibodies and small molecules (see General Methods) in the Tango™ (Thermo) β-arrestin recruitment assay to CXCR2 using human CXCL1, CXCL5 and CXCL8 ligands at calculated values of EC ₅₀ .

BKO-4A8-101c inhibited CXCR2-mediated β-arrestin functional activity induced by a panel of known CXCR2 ligands, with comparable IC ₅₀ values for the different ligands. The antibody exhibited ligand-selective inhibition of CXCR2-dependent β-arrestin recruitment, with complete inhibition of CXCL1- and CXCL5-induced and partial inhibition of CXCL8-induced β-arrestin recruitment (range 70-80%), as shown in Table 1. 17.

The efficiency of BKO-4A8-101c inhibition of CXCL1 and CXCL5-mediated β-arrestin signaling was similar to or greater than that observed for other comparator antibodies and small molecules. Based on _IC50 values, BKO-4A8-101c was found to be 2- to 39-fold more effective in inhibiting CXCL1 or CXCL5 ligand-induced β-arrestin activation, but did not completely inhibit CXCL8-induced CXCR2 activation. The incomplete inhibition of CXCL8-mediated β-arrestin reporter activity suggests that the β-arrestin-mediated receptor internalization pathway is functional.

Table 17. Summary of mean IC ₅₀ values of CXCR2 antagonists for the effect of human ELR+CXC chemokines on human CXCR2 in β-arrestin recruitment assay

Compound CXCL1 CXCL5 CXCL8 IC ₅₀ (nM) max.inhibition (%) IC ₅₀ (nM) max.inhibition (%) IC ₅₀ (nM) max.inhibition (%) BKO-4A8-101c 0.38 97 0.34 98 0.34 78 Antagonist 1 1.20 98 0.80 98 0.58 97 Antagonist 2 1.14 93 0.56 96 0.82 74 Antagonist 3 14.80 85 8.12 96 n/ ^a 82 Antagonist 4 1.47 96 0.75 99 1.01 94 Antagonist 5 2.77 99 1.75 99 1.23 99 Antagonist 6 0.71 99 0.29 97 1.22 98

^a Does not inhibit or the data are not suitable for constructing a four-point dose-response curve

Characteristics of BKO-4A8-101c inhibition of calcium mobilization

The ability of antagonists to inhibit CXCR2 activation of calcium mobilization induced by CXCL1 and CXCL8 was assessed in the commercially available calcium-optimized human chemokine receptor CXCR2 stable cell line HTS002C-Chemiscreen™ (Eurofins Pharma Discovery Services). CXC ligands were added to the samples at a concentration of EC ₅₀ (see Table 3) for the analysis of antagonists. To measure calcium mobilization in cells, a Fluo-4 NW calcium assay kit (Life Technologies) was used according to the manufacturer's procedure and read on a FLIPR Tetra high-throughput cell screen (Molecular Devices). The peak signal minus the basal signal from each well was used to construct dose-response inhibition curves using a 4-parameter logistic equation and determine IC ₅₀ values.

The ability of BKO-4A8-101c to block ligand-induced calcium influx was compared with comparative antibodies and small molecules (Table 18) using human CXCL1 and CXCL8 ligands at calculated EC ₅₀ values. In table 18 shows IC ₅₀ values from at least 3 independent replicates. BKO-4A8-101c was an equivalent or more potent CXCL1 antagonist than comparator antibodies and small molecules and potently inhibited human CXCL1-induced calcium influx. In contrast, BKO-4A8-101c and other comparative antibodies showed little or no inhibition of CXCL8-induced calcium influx, whereas comparative small molecule CXCR2 antagonists were able to completely inhibit CXCL8-induced calcium influx in this assay. The CXCR2-mediated neutrophil chemotactic response is dependent on calcium mobilization. Without committing to any proposed mechanism of action, the selective antagonistic activity observed in BKO-4A8-101c provides a therapeutic window to virtually completely inhibit CXCL1- and CXCL5-mediated neutrophil migration into tissues without significantly affecting CXCL8-mediated migration from the bone marrow. This ligand selectivity may also influence chemokine gradients that direct neutrophil chemotactic responses.

Table 18. Summary of IC ₅₀ values of human CXCR2 antagonists for the effect of human ELR+CXC chemokines on human CXCR2 in a calcium influx assay

Compound CXCL1 CXCL8 IC ₅₀ (nM) Max. inhibition (%) IC ₅₀ (nM) Max. inhibition (%) BKO-4A8-101c 1.47 93 n/ ^a 9 Antagonist 1 n/ ^a 28 n/ ^a 8 Antagonist 2 1.02 79 n/ ^a 3 Antagonist 3 30.74 37 n/ ^a 18 Antagonist 4 1.29 93 n/ ^a 1 Antagonist 5 48.94 95 115.12 97 Antagonist 6 8.46 96 16.09 98

^a Does not inhibit or the data are not suitable for constructing a four-point dose-response curve

Example 10 Anti-CXCR2 Antibody Inhibits Lung Neutrophilia in a Mouse Model of Severe Asthma

Sensitized mice following intranasal administration of a challenge dose of house dust mite (HDM) extract develop an inflammatory profile with mixed pulmonary eosinophilia and neutrophilia along with goblet cell hyperplasia. A model of severe asthma using SCH527123, a small molecule CXCR2 antagonist, has been previously reported at doses of 10 and 30 mg/kg (discussed in Young A. et al. The Effect of the CXCR1/2 Antagonist SCH257123 in a Mouse Model of Severe Asthma. Experimental Biology 2016 Meeting, 2016 San Diego, USA: The FASEB Journal, 1202.10), which works in mouse models of neutrophil inflammation (as discussed in Chapman R.W. et al. A novel, orally active CXCR1/2 receptor antagonist, Sch527123, inhibits neutrophil recruitment, mucus production , and goblet cell hyperplasia in animal models of pulmonary inflammation. J Pharmacol Exp Ther. (2007) 322, 486-93). Pretreatment with 10 and 30 mg/kg SCH527123 only suppressed neutrophil cell counts by a maximum of 30% in BAL, which was not significantly different from the house dust mite vehicle (HDM) group (as discussed in Young et al., supra).

The anti-CXCR2 antibody BKO-4A8-mIgG1 (SEQ ID NOs: 113 and 114) was generated by formatting the heavy and light chain variable regions of BKO-4A8 to the IgG1 constant region of mice with reduced effector function. Female hCXCR2-introduced mice were sensitized with 50 μg HDM in complete Freund's adjuvant (CFA) administered subcutaneously on day 0 and an intranasal boost with 50 mg HDM without CFA on day 14. Animals were treated with vehicle or BKO-4A8-mIgG1 (10 mg/kg) by intraperitoneal injection on days 5 and 12. Inflammatory responses were monitored at the endpoint of day 16 by total and differential cell counts in the bronchoalveolar lavage fluid (BAL) of the right lung. The left lung was fixed in 10% formalin for histopathology and assessment of mucus production.

Treatment with the anti-CXCR2 antibody BKO-4A8-mIgG1 prior to HDM challenge resulted in a reduction in disease severity, including a significant (>60%) reduction in eosinophils (p = 0.017) and neutrophils in BAL and a reduction in goblet cell hyperplasia (p = 0.0052) by compared to the vehicle-treated control group, as seen in FIG. 13A, 13B and 13C. To identify statistically significant differences between vehicle and treatment groups, statistical analysis was performed using a nonparametric unpaired Mann-Whitney t test and outliers were identified using Grubbs' test (alpha = 0.01). All statistical analyzes were performed using GraphPad Prism™ 7.01.

The reduction in BAL eosinophil numbers in this model was unexpected. The ability of BKO-4A8-mIgG1 to inhibit eosinophil migration supports its utility in the treatment of eosinophilic diseases such as eosinophilic asthma, along with neutrophilic diseases.

Example 11: LPS-Induced Acute Pneumonia in Cynomolgus Macaques

Biologic-naïve (i.e., not previously treated with exogenous biologics) male cynomolgus monkeys were randomly assigned to receive vehicle or anti-CXCR2 antibody BKO-4A8-101c intravenously at a dose of 1 mg/kg on days 0, 14, and 28. 1 hour after administration of test antibodies on day 0, animals were exposed to aerosolized bacterial lipopolysaccharide (LPS) by inhalation of 20 μg/L over 5 min (total dose 20 μg/kg). This is a well-validated model of acute pneumonia. At 24 hours after LPS exposure, inflammatory responses were monitored by total and differential cell counts in left lung bronchoalveolar lavage fluid and compared with corresponding counts in naïve animals on day 14 before treatment. At various time points, blood was drawn for differential cell counts and serum was collected for pharmacokinetic analysis.

A single treatment with LPS aerosol induced an influx of neutrophils into the lungs of vehicle-treated animals. Pretreatment with 1 mg/kg anti-CXCR2 antibody BKO-4A8-101c markedly inhibited LPS-induced lung neutrophilia in cynomolgus monkeys, as seen in FIG. 14A. The treatment was well tolerated without loss of body weight. As can be seen from Fig. 14B, treatment with BKO-4A8-101c did not cause significant changes in the number of neutrophils in the blood after repeated administration of antibodies.

CXCR2 signaling is involved in the trafficking of neutrophils both from the bone marrow and into peripheral tissues in response to chemokines produced by tissue resident cells following stress, injury, or infection. CXCR2 binds to several chemokines involved in neutrophil recruitment and chronic inflammation, including CXCL1, CXCL5, and CXCL8. These chemokines are also elevated in patients with severe neutrophilic asthma and COPD. Here, BKO-4A8-101c was shown to be a potent and specific antagonist of CXCR2-mediated signaling. Without adhering to any proposed mechanism of action, based on its in vitro profile and efficacy in cynomolgus monkeys, the anti-inflammatory activity of BKO-4A8-101c appears to be mediated by antagonism of CXCL1 and CXCL5-mediated CXCR2 signaling. These data support the concept that the CXCR2 receptor is the predominant chemokine receptor controlling neutrophil migration into the lung under inflammatory conditions and are consistent with the lack of detectable efficacy of a humanized neutralizing antibody against CXCL8 when administered to patients with COPD (as discussed in Mahler D.A. et al. Efficacy and safety of a monoclonal antibody recognizing interleukin-8 in COPD: a pilot study. Chest (2004) 126, 926-34), an approach that only partially suppresses the CXCR1/CXCR2 inflammatory axis. Some component of the anti-inflammatory activity of BKO-4A8-101c may be mediated through endothelial and epithelial cells, since CXCR2 expression in these cell types is also involved in neutrophil recruitment and lung injury (as discussed in Reutershan J. et al. Critical role of endothelial CXCR2 in LPS-induced neutrophil migration into the lung. J. Clin. Invest. (2006) 116, 695-702). The demonstrated in vivo efficacy of BKO-4A8-101c in inhibiting neutrophil migration into the lung in response to LPS stimulation, without affecting circulating neutrophil counts or any other safety parameters measured, may be a consequence of its high specificity and selective antagonistic activity.

Example 12 Anti-CXCR2 Antibody Effectively Occupies the CXCR2 Receptor and Selectively Inhibits CXCR2-Mediated Neutrophil Responses

Receptor occupancy assays measure the binding of a specific molecule or drug to a receptor expressed on specific cells. This is a quantitative assay that can be used to evaluate receptor binding and other pharmacodynamic properties.

Receptor occupancy was examined in a cell line expressing human CXCR2 and human neutrophils enriched from whole blood. At least 85% of CXCR2 receptors were occupied at 2 μg/ml antibody. This amount was sufficient to suppress CXCL1-induced calcium influx by more than 85% in HTS002C-Chemiscreen™ cells with calcium-optimized human chemokine receptor CXCR2 (results not shown).

Although 2 μg/mL of antibody was sufficient to effectively antagonize CXCR2-mediated signaling, it did not affect neutrophil function. The final targets of the chemoattractants C5a and fMLF bind to the C5a receptor (CD88) and FPR1, respectively. Both of these agents induce chemotaxis and expression of CD11b, a common marker of neutrophil activation, in response to infection and inflammation. Antibody to CXCR2 did not suppress the increase in neutrophil CD11b levels or chemotaxis in response to C5a and fMLF (results not shown).

Anti-CXCR2 antibody at 10 μg/ml (70 nM) effectively and specifically antagonized CXCR2-mediated responses to CXCL1 (p < 0.0002, Figure 15A) and CXCL5 (p < 0.0001, Figure 15B) in human neutrophils isolated (up to 95%) from whole blood. Anti-CXCR2 antibody at 70 nM was more potent in this assay than small molecule antagonist 5 (danirixin) at 1400 nM. This is consistent with previous reports that agonist 5 inhibits ex vivo CXCL1-induced CD11b expression on the surface of neutrophils with _IC50 = 69 ng/ml [156 nM] and _IC90 = 620 ng/ml (range 158-1080 ng/ml) [1400 nM, range 356-2443 nM] (Miller et al. “The pharmacokinetics and pharmacodynamics of danirixin (GSK1325756) - a selective CXCR2 antagonist - in healthy adult subjects” BMC Pharmacology and Toxicology 2015, 16).

Neither CXCR2 nor antagonist 5 significantly affected the neutrophil response to CXCL8 in this assay (Figure 15C). CXCL8 binds to both CXCR1 and CXCR2.

These data indicate that anti-CXCR2 antibody is a potent and selective inhibitor of CXCR2 on human neutrophils.

Those skilled in the art will appreciate that various changes and modifications can be made to the preferred embodiments of the invention, and such changes and modifications can be made without departing from the spirit of the invention. It is therefore intended that the appended claims cover all such equivalent embodiments as are consistent with the true spirit of the invention and fall within the scope thereof.

The contents of each patent, patent application and publication cited or described herein are hereby incorporated by reference in their entirety.

Incarnations

The following list of embodiments is intended to complement, and not to supersede or replace, the preceding description.

Embodiment 1. A human antibody molecule that immunospecifically binds to human CXCR2, the antibody molecule comprising:

Heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 167 and light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 168; or

Heavy chain CDR1, CDR2 and CDR3 of SEQ ID NO: 226 and light chain CDR1, CDR2 and CDR3 of SEQ ID NO: 227;

and wherein the antibody molecule inhibits CXCL1-induced activation of CXCR2 or CXCL5-induced activation of CXCR2.

Embodiment 2. The human antibody molecule from Embodiment 1, wherein:

Heavy chain CDR1 has the amino acid sequence of SEQ ID NO: 169, heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 170, heavy chain CDR3 has the amino acid sequence of SEQ ID NO: 171, light chain CDR1 has the amino acid sequence of SEQ ID NO: 172, the light chain CDR2 has the amino acid sequence of SEQ ID NO: 173, and the light chain CDR3 has the amino acid sequence of SEQ ID NO: 174; or

Heavy chain CDR1 has the amino acid sequence of SEQ ID NO: 228, heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 229, heavy chain CDR3 has the amino acid sequence of SEQ ID NO: 230, light chain CDR1 has the amino acid sequence of SEQ ID NO: 201, the light chain CDR2 has the amino acid sequence of SEQ ID NO: 231, and the light chain CDR3 has the amino acid sequence of SEQ ID NO: 232.

Embodiment 3. The human antibody molecule from embodiment 1 or 2, wherein:

the heavy chain variable region has the amino acid sequence of SEQ ID NO: 167, and the light chain variable region has the amino acid sequence of SEQ ID NO: 168; or

the heavy chain variable region has the amino acid sequence of SEQ ID NO: 226, and the light chain variable region has the amino acid sequence of SEQ ID NO: 227.

Embodiment 4. The human antibody molecule from Embodiment 3, wherein:

a) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 98, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

b) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 99, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

c) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 100, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

d) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 101, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

e) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 102, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

f) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 103, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

g) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 104, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

h) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 105 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

i) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 106, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

j) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 107, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

k) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

l) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 162, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

m) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 163, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

n) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 164, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110;

o) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 165, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; or

p) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 166, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110.

Embodiment 5. The human antibody molecule from Embodiment 4, wherein:

a) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110;

b) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 162, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110;

c) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 163, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110;

d) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 164, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110;

e) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 165, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; or

f) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 166, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110.

Embodiment 6. The human antibody molecule of Embodiment 1 or 2, wherein the heavy chain CDR1 has the amino acid sequence of SEQ ID NO: 182, the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 192, the heavy chain CDR3 has the amino acid sequence of SEQ ID NO: : 195, light chain CDR1 has the amino acid sequence of SEQ ID NO: 201, light chain CDR2 has the amino acid sequence of SEQ ID NO: 205, and light chain CDR3 has the amino acid sequence of SEQ ID NO: 213.

Embodiment 7. The human antibody molecule of any one of embodiments 1 to 6, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108 and the light chain variable region has the amino acid sequence of SEQ ID NO: 110.

Embodiment 8. The human antibody molecule of any one of embodiments 1-7, wherein the antibody comprises a human IgG1 heavy chain constant region.

Embodiment 9. The human antibody molecule of Embodiment 8, wherein the human IgG1 heavy chain constant region has the amino acid sequence of SEQ ID NO: 122 or 124.

Embodiment 10. The human antibody molecule of any one of embodiments 1-7, wherein the antibody comprises a human IgG2 heavy chain constant region.

Embodiment 11. The human antibody molecule of Embodiment 10, wherein the human IgG2 heavy chain constant region has the amino acid sequence of SEQ ID NO: 120.

Embodiment 12. The human antibody molecule of any one of embodiments 1-7, wherein the antibody comprises a human IgG4 heavy chain constant region.

Embodiment 13. The human antibody molecule of any one of embodiments 1-12, wherein the antibody molecule is a Fab fragment, an F(ab) ₂ fragment, or a single chain antibody.

Embodiment 14. A pharmaceutical composition containing human antibody molecules according to any of embodiments 1-13.

Embodiment 15. A nucleic acid molecule encoding a human antibody molecule according to any one of embodiments 1-13.

Embodiment 16. A vector containing the nucleic acid molecule from embodiment 15.

Embodiment 17. A cell transformed to express a human antibody molecule according to any one of embodiments 1-13.

Embodiment 18. A method for treating or preventing airway neutrophilia or acute pneumonia in a subject, which comprises:

administering to a subject a therapeutically effective amount of the human antibody molecule of any one of embodiments 1-13 or the pharmaceutical composition of embodiment 14 to prevent or treat airway neutrophilia or acute pneumonia.

Embodiment 19. The method of embodiment 18, wherein the airway neutrophilia or acute pneumonia or both is chronic obstructive pulmonary disease, severe neutrophilic asthma or both.

Embodiment 20. The human antibody molecule of any one of embodiments 1-13 or the pharmaceutical composition of embodiment 14 for use as a medicine.

Embodiment 21. The human antibody molecule of any one of embodiments 1-13 or the pharmaceutical composition of embodiment 14 for use in the prevention or treatment of airway neutrophilia or acute pneumonia.

Embodiment 22. Use of the human antibody molecules of any of embodiments 1-13 or the pharmaceutical composition of embodiment 14 in the preparation of a medicament for the prevention or treatment of respiratory neutrophilia or acute pneumonia.

Embodiment 23. The use of embodiment 21 or 22, wherein the airway neutrophilia, acute pneumonia, or both is chronic obstructive pulmonary disease, severe neutrophilic asthma, or both.

Embodiment 24. A method for blocking neutrophil chemotaxis, comprising exposing neutrophils to a human antibody according to any of embodiments 1-13.

Embodiment 25. The method of embodiment 24, wherein chemotaxis is the migration of neutrophils into the lungs.

Embodiment 26. A method of blocking CXCR2 signaling in response to CXCL1 and/or CXCL5 in cells expressing CXCR2 signaling, comprising exposing the cells to a human antibody according to any of embodiments 1-13.

--->

LIST OF SEQUENCES

<110> CEPHALON, INC.

<120> ANTI-CXCR2 ANTIBODIES AND USES THEREOF

<130> 102085.005304

<140>

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Glu Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Lys Trp Val

35 40 45

Ser Gly Ile Thr Trp Asn Ser Gly Asn Lys Arg Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95

Ala Lys Asp Met Lys Gly Ser Gly Thr Tyr Phe Pro Ala Phe Asp Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 14

<211> 106

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_3D6_L6_G06_VL (BKO_5H4_VL)"

<400> 14

Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp Lys Tyr Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Val Val

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 15

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_3F4_VH"

<400> 15

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Lys Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Val Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 16

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_3F4_L11_A11_VL"

<400> 16

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Met Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Thr Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Ala Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Pro

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 17

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_4A8_VH"

<400> 17

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 18

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_4A8_VL"

<400> 18

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 19

<211> 123

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_4F10_VH"

<400> 19

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Phe Asn Pro Asn Asn Gly Gly Thr Asn Tyr Ala Gln Arg Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Pro Thr Ile Arg Leu Trp Phe Asp Asn Trp Phe Asp Ser

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 20

<211> 110

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_4F10_VL"

<400> 20

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Ile Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 21

<211> 122

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5E8_H5_C05_VH"

<400> 21

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Arg Gly Ser Gly Ala Gly Thr Tyr Tyr Ala Asp Ser Met

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr

65 70 75 80

Leu Leu Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ser Lys Leu Glu Ala Val Ser Gly Thr Gly Lys Tyr Phe Gln His Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 22

<211> 106

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5E8_L3_C03_VL"

<400> 22

Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Asn Ile Thr Cys Ser Gly Asp Thr Leu Gly Asp Lys Phe Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Thr Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Lys Ser Gly Ile Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Phe Tyr Cys Gln Ala Trp Asn Ser Arg Gly Val Val

85 90 95

Phe Gly Gly Gly Thr Arg Leu Thr Val Leu

100 105

<210> 23

<211> 112

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5G11_VH"

<400> 23

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

100 105 110

<210> 24

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5G11_VL"

<400> 24

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Phe Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 25

<211> 123

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5G6_VH"

<400> 25

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Thr Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Phe Asn Pro Asn Asn Gly Gly Thr Asn Tyr Ala Gln Arg Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Pro Thr Ile Arg Leu Trp Phe Asp Asn Trp Phe Asp Ser

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 26

<211> 110

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_5G6_L12_E12_VL"

<400> 26

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Ile Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Trp Val Phe Gly Gly Gly Thr Thr Leu Thr Val Leu

100 105 110

<210> 27

<211> 123

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_6A1_H4_A04_VH"

<400> 27

Gln Val Gln Leu Lys Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Gly Glu Val Arg Gly Leu Ile Thr Leu Tyr Trp Tyr Phe Asp Val

100 105 110

Trp Gly Arg Gly Ser Leu Val Thr Val Ser Ser

115 120

<210> 28

<211> 108

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_6A1_E10_VL"

<400> 28

Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala

20 25 30

Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr

35 40 45

Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His

85 90 95

Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 29

<211> 120

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_6A2_H1_B01_VH"

<400> 29

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Glu Gly Tyr Asn Tyr Gly Tyr Gly Gly Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 30

<211> 106

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_6A2_L6_A06_VL"

<400> 30

Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp Lys Tyr Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Val Val

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 31

<211> 123

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7C11_H6_B06_VH"

<400> 31

Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Ser Arg Asn Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Gly Glu Val Arg Gly Val Phe Thr Leu Tyr Trp Tyr Phe Asp Val

100 105 110

Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 32

<211> 108

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7C11_G01_VL"

<400> 32

Ser Ser Glu Leu Thr Gln Gly Pro Ala Val Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Arg Ile Thr Cys Gln Gly Asn Ser Leu Arg Phe Tyr Tyr Ala

20 25 30

Ser Trp Tyr Gln Gln Arg Pro Gly Gln Ala Pro Ile Leu Val Ile Tyr

35 40 45

Asp Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Tyr Tyr

85 90 95

Met Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 33

<211> 112

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7G10_H1_B01_VH"

<400> 33

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

100 105 110

<210> 34

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7G10_L6_E06_VL"

<400> 34

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Phe Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 35

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7H8_H3_C03_VH"

<400> 35

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Ala Ile Ser Gly Arg Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Leu Gly Asn Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 36

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_7H8_L10_F10_VL"

<400> 36

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Val Ile Tyr Glu Val Asn Met Arg Pro Ser Gly Val Pro Ala Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asp Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Ser Val Leu

100 105 110

<210> 37

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8B6_VH"

<400> 37

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Asn

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Asn Ser Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Leu Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Lys Leu Gly Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser

100 105 110

Ser

<210> 38

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8B6_VL"

<400> 38

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Met Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asp Asn Phe Gly Val Phe Gly Gly Gly Thr Arg Leu Thr Val Leu

100 105 110

<210> 39

<211> 122

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8C4_VH"

<400> 39

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Arg Thr Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Tyr Ile Gly Ser Ile Tyr Tyr Ser Gly Thr Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg His Gly Arg Val Arg Glu Val Pro Pro Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 40

<211> 108

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8C4_VL"

<400> 40

Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Tyr Tyr Tyr Ala

20 25 30

Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr

35 40 45

Asp Glu Asn Ser Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Ser Ser Gly Asn Thr Ala Ser Leu Arg Ile Thr Gly Thr Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Thr Ser Gly Asn His

85 90 95

Trp Ala Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 41

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8G3_H4_D04_VH"

<400> 41

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Thr Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Arg Leu Gly Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser

100 105 110

Ser

<210> 42

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8G3_L1_G01_VL"

<400> 42

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Val Pro Lys Leu

35 40 45

Val Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 43

<211> 116

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8H10_VH"

<400> 43

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Lys Pro Asp Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Ser Gly Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 44

<211> 110

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8H10_VL"

<400> 44

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Phe Thr Gly Thr Ser Arg Asp Val Gly Asp Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Thr Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 45

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8H8_H5_E05_VH"

<400> 45

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Val Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Lys Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Ser Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Val Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 46

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_8H8_L7_H08_VL"

<400> 46

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Met Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Thr Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Val Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Val Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Pro

85 90 95

Asn Asn Phe Gly Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 47

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_9A8_H3_F03_VH"

<400> 47

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Asn

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Arg Thr Tyr Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met His Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Leu Gly Ser Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 48

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_9A8_L1_H02_VL"

<400> 48

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ala Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 49

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_9C3_H8_G08_VH"

<400> 49

Glu Val Gln Val Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Leu Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 50

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_9C3_L1_F01_VL"

<400> 50

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Ile Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Val Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Thr Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 51

<211> 119

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_10G10_VH"

<400> 51

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Arg Arg Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Phe Tyr Asn Ser Gly Asn Thr Tyr Tyr Lys Pro Ser

50 55 60

Leu Lys Ser Arg Val Ala Ile Ser Val Asp Thr Pro Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Gly Tyr Ser Ser Gly Gly Phe Asp Pro Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 52

<211> 110

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO_10G10_VL"

<400> 52

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Cys Gln Gln His Pro Gly Lys Ala Pro Lys Ile

35 40 45

Met Ile Phe Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Trp Val Phe Gly Gly Gly Thr Arg Leu Thr Val Leu

100 105 110

<210> 53

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32Q"

<400> 53

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Gln

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 54

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32H"

<400> 54

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 55

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32L"

<400> 55

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Leu

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 56

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32W"

<400> 56

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Trp

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 57

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32Y"

<400> 57

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 58

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH T33A"

<400> 58

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 59

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M34Q"

<400> 59

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 60

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M34D"

<400> 60

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Asp Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 61

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M34H"

<400> 61

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr His Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 62

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M34W"

<400> 62

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Trp Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 63

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH I51H"

<400> 63

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala His Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 64

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH G52aD"

<400> 64

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Asp Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 65

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH R53S"

<400> 65

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 66

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH R53Q"

<400> 66

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Gln Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 67

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH G54D"

<400> 67

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Asp Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 68

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH N56S"

<400> 68

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 69

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH I94K"

<400> 69

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 70

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96A"

<400> 70

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Ala Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 71

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96Q"

<400> 71

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Gln Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 72

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96K"

<400> 72

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 73

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH G101D"

<400> 73

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Asp Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 74

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Y102S"

<400> 74

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Ser Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 75

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Y102K"

<400> 75

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Lys Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 76

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL E50D"

<400> 76

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 77

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N52D"

<400> 77

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asp Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 78

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N52S"

<400> 78

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 79

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL K53A"

<400> 79

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Ala Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 80

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL K53D"

<400> 80

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Asp Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 81

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL K53H"

<400> 81

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn His Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 82

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL R54Q"

<400> 82

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Gln Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 83

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL Y91A"

<400> 83

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Ala Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 84

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94A"

<400> 84

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ala

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 85

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94S"

<400> 85

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 86

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94K"

<400> 86

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Lys

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 87

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94L"

<400> 87

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Leu

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 88

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94W"

<400> 88

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Trp

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 89

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94Y"

<400> 89

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Tyr

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 90

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aQ"

<400> 90

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Gln Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 91

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aD"

<400> 91

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asp Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 92

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aH"

<400> 92

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn His Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 93

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aK"

<400> 93

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Lys Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 94

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aL"

<400> 94

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Leu Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 95

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aY"

<400> 95

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Tyr Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 96

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V97A"

<400> 96

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Ala Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 97

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V97K"

<400> 97

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Lys Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 98

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 1 (M34Q_N56S)"

<400> 98

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 99

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 2 (M34Q_A40P_N56S)"

<400> 99

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 100

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 3 (M34Q_A40P_N56S_R75K)"

<400> 100

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 101

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 4 (M34Q_A40P_N56S_M96K)"

<400> 101

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 102

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 5 (M34Q_A40P_N56S_R75K_M96K)"

<400> 102

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 103

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 6 (M34Q_A40P_N56S_R75K_I94K_M96K)"

<400> 103

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 104

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 7 (M34Q_A40P_N56S_I94K_M96K)"

<400> 104

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 105

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 8 (M34Q_N56S_M96K)"

<400> 105

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 106

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 9 (M34Q_N56S_R75K_M96K)"

<400> 106

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 107

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 10 I94K_M96K"

<400> 107

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 108

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 101 M34Q_A40P_N56S_R75K_M96A"

<400> 108

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Ala Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 109

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL variant b N52S_N94S"

<400> 109

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 110

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL variant with D41G_N52S_N94S"

<400> 110

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 111

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="4A8 VH M34Q_A40P_N56S_R75K_M96A"

<400> 111

gaagttcagc tgcttgaatc tggcggagga ctggttcagc ctggcggatc tctgagactg

60

tcttgtgccg ccagcggctt cacctttagc agcagcacac agagctgggt ccgacagcct

120

cctggcaaag gactggaatg ggtgtccgcc atctctggca gaggcagaag cacctactac

180

gccgactctg tgaagggcag attcaccatc agccgggaca acagcaagaa caccctgtac

240

ctgcagatga acagcctgag agccgaggac accgccgtgt actattgtgc catccaggcc

300

ggctattggg gccagggaat actcgtgaca gtgtcctca

339

<210> 112

<211> 333

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="4A8 VL D41G_N52S_N94S"

<400> 112

cagtctgctc tgacacagcc tcctagcgcc tctggctctc ctggccagag cgtgaccatc

60

agctgtatcg gcaccagcag cgacgtgggc ggctacaact acgtgtcctg gtatcagcag

120

caccccggta aggcccccaa gctgatgatc tacgaagtgt ccaagcggcc cagcggcgtg

180

cccgatagat tcagcggcag caagagcggc aacaccgcca gcctcacagt gtctggactg

240

caggccgagg acgaggccga ctactactgt agcagctacg ccggcagcaa caacttcggc

300

gtgttcggcg gaggcaccaa gctgacagtc cta

333

<210> 113

<211> 436

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8-mIgG1 VH"

<400> 113

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser

115 120 125

Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu Val Lys Gly

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr

165 170 175

Leu Ser Ser Ser Val Thr Val Pro Ser Ser Pro Arg Pro Ser Glu Thr

180 185 190

Val Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys

195 200 205

Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val

210 215 220

Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val

225 230 235 240

Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys Val Val Val Ala Ile

245 250 255

Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp Phe Val Asp Asp Val

260 265 270

Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser

275 280 285

Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met His Gln Asp Trp Leu

290 295 300

Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala

305 310 315 320

Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro

325 330 335

Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln Met Ala Lys Asp Lys

340 345 350

Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr

355 360 365

Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr

370 375 380

Gln Pro Ile Met Asn Thr Asn Gly Ser Tyr Phe Val Tyr Ser Lys Leu

385 390 395 400

Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser

405 410 415

Val Leu His Glu Gly Leu His Asn His His Thr Glu Lys Ser Leu Ser

420 425 430

His Ser Pro Gly

435

<210> 114

<211> 217

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8-mIgG1 VL"

<400> 114

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

100 105 110

Gln Pro Lys Ser Ser Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu

115 120 125

Glu Leu Glu Thr Asn Lys Ala Thr Leu Val Cys Thr Ile Thr Asp Phe

130 135 140

Tyr Pro Gly Val Val Thr Val Asp Trp Lys Val Asp Gly Thr Pro Val

145 150 155 160

Thr Gln Gly Met Glu Thr Thr Gln Pro Ser Lys Gln Ser Asn Asn Lys

165 170 175

Tyr Met Ala Ser Ser Tyr Leu Thr Leu Thr Ala Arg Ala Trp Glu Arg

180 185 190

His Ser Ser Tyr Ser Cys Gln Val Thr His Glu Gly His Thr Val Glu

195 200 205

Lys Ser Leu Ser Arg Ala Asp Cys Ser

210 215

<210> 115

<211> 439

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8 IgG4"

<400> 115

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

115 120 125

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys

180 185 190

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala

210 215 220

Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

225 230 235 240

Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val

245 250 255

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

260 265 270

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

275 280 285

Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

290 295 300

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

305 310 315 320

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

325 330 335

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr

340 345 350

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

355 360 365

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

370 375 380

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

385 390 395 400

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

405 410 415

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

420 425 430

Ser Leu Ser Leu Ser Leu Gly

435

<210> 116

<211> 326

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="IgG4"

<400> 116

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly

325

<210> 117

<211> 439

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8 IgG4"

<400> 117

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

115 120 125

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys

180 185 190

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala

210 215 220

Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

225 230 235 240

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

245 250 255

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

260 265 270

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

275 280 285

Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

290 295 300

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

305 310 315 320

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

325 330 335

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr

340 345 350

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

355 360 365

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

370 375 380

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

385 390 395 400

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

405 410 415

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

420 425 430

Ser Leu Ser Leu Ser Leu Gly

435

<210> 118

<211> 326

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="IgG4"

<400> 118

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly

325

<210> 119

<211> 438

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8 IgG2"

<400> 119

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

115 120 125

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln

180 185 190

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala

210 215 220

Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

225 230 235 240

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

245 250 255

Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

260 265 270

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

275 280 285

Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp

290 295 300

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

305 310 315 320

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg

325 330 335

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys

340 345 350

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

355 360 365

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

370 375 380

Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

385 390 395 400

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

405 410 415

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

420 425 430

Leu Ser Leu Ser Pro Gly

435

<210> 120

<211> 325

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="IgG2"

<400> 120

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

100 105 110

Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

115 120 125

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

130 135 140

Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

145 150 155 160

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

165 170 175

Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp

180 185 190

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

195 200 205

Ser Ser Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu

210 215 220

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn

225 230 235 240

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

245 250 255

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

260 265 270

Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

275 280 285

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

290 295 300

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

305 310 315 320

Ser Leu Ser Pro Gly

325

<210> 121

<211> 442

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8 IgG1"

<400> 121

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

115 120 125

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

180 185 190

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

210 215 220

Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro

225 230 235 240

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

245 250 255

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

260 265 270

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

275 280 285

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

290 295 300

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

305 310 315 320

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

325 330 335

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

340 345 350

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

355 360 365

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

370 375 380

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

385 390 395 400

Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

405 410 415

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

420 425 430

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440

<210> 122

<211> 329

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="IgG1"

<400> 122

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 123

<211> 442

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="BKO-4A8 IgG1"

<400> 123

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

115 120 125

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

180 185 190

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

210 215 220

Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro

225 230 235 240

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

245 250 255

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

260 265 270

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

275 280 285

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

290 295 300

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

305 310 315 320

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

325 330 335

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

340 345 350

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

355 360 365

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

370 375 380

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

385 390 395 400

Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

405 410 415

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

420 425 430

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440

<210> 124

<211> 329

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="IgG1"

<400> 124

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 125

<211> 360

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="Human CXCR2"

<400> 125

Met Glu Asp Phe Asn Met Glu Ser Asp Ser Phe Glu Asp Phe Trp Lys

1 5 10 15

Gly Glu Asp Leu Ser Asn Tyr Ser Tyr Ser Ser Thr Leu Pro Pro Phe

20 25 30

Leu Leu Asp Ala Ala Pro Cys Glu Pro Glu Ser Leu Glu Ile Asn Lys

35 40 45

Tyr Phe Val Val Ile Ile Tyr Ala Leu Val Phe Leu Leu Ser Leu Leu

50 55 60

Gly Asn Ser Leu Val Met Leu Val Ile Leu Tyr Ser Arg Val Gly Arg

65 70 75 80

Ser Val Thr Asp Val Tyr Leu Leu Asn Leu Ala Leu Ala Asp Leu Leu

85 90 95

Phe Ala Leu Thr Leu Pro Ile Trp Ala Ala Ser Lys Val Asn Gly Trp

100 105 110

Ile Phe Gly Thr Phe Leu Cys Lys Val Val Ser Leu Leu Lys Glu Val

115 120 125

Asn Phe Tyr Ser Gly Ile Leu Leu Leu Ala Cys Ile Ser Val Asp Arg

130 135 140

Tyr Leu Ala Ile Val His Ala Thr Arg Thr Leu Thr Gln Lys Arg Tyr

145 150 155 160

Leu Val Lys Phe Ile Cys Leu Ser Ile Trp Gly Leu Ser Leu Leu Leu

165 170 175

Ala Leu Pro Val Leu Leu Phe Arg Arg Thr Val Tyr Ser Ser Asn Val

180 185 190

Ser Pro Ala Cys Tyr Glu Asp Met Gly Asn Asn Thr Ala Asn Trp Arg

195 200 205

Met Leu Leu Arg Ile Leu Pro Gln Ser Phe Gly Phe Ile Val Pro Leu

210 215 220

Leu Ile Met Leu Phe Cys Tyr Gly Phe Thr Leu Arg Thr Leu Phe Lys

225 230 235 240

Ala His Met Gly Gln Lys His Arg Ala Met Arg Val Ile Phe Ala Val

245 250 255

Val Leu Ile Phe Leu Leu Cys Trp Leu Pro Tyr Asn Leu Val Leu Leu

260 265 270

Ala Asp Thr Leu Met Arg Thr Gln Val Ile Gln Glu Thr Cys Glu Arg

275 280 285

Arg Asn His Ile Asp Arg Ala Leu Asp Ala Thr Glu Ile Leu Gly Ile

290 295 300

Leu His Ser Cys Leu Asn Pro Leu Ile Tyr Ala Phe Ile Gly Gln Lys

305 310 315 320

Phe Arg His Gly Leu Leu Lys Ile Leu Ala Ile His Gly Leu Ile Ser

325 330 335

Lys Asp Ser Leu Pro Lys Asp Ser Arg Pro Ser Phe Val Gly Ser Ser

340 345 350

Ser Gly His Thr Ser Thr Thr Leu

355 360

<210> 126

<211> 359

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="Mouse CXCR2"

<400> 126

Met Gly Glu Phe Lys Val Asp Lys Phe Asn Ile Glu Asp Phe Phe Ser

1 5 10 15

Gly Asp Leu Asp Ile Phe Asn Tyr Ser Ser Gly Met Pro Ser Ile Leu

20 25 30

Pro Asp Ala Val Pro Cys His Ser Glu Asn Leu Glu Ile Asn Ser Tyr

35 40 45

Ala Val Val Val Ile Tyr Val Leu Val Thr Leu Leu Ser Leu Val Gly

50 55 60

Asn Ser Leu Val Met Leu Val Ile Leu Tyr Asn Arg Ser Thr Cys Ser

65 70 75 80

Val Thr Asp Val Tyr Leu Leu Asn Leu Ala Ile Ala Asp Leu Phe Phe

85 90 95

Ala Leu Thr Leu Pro Val Trp Ala Ala Ser Lys Val Asn Gly Trp Thr

100 105 110

Phe Gly Ser Thr Leu Cys Lys Ile Phe Ser Tyr Val Lys Glu Val Thr

115 120 125

Phe Tyr Ser Ser Val Leu Leu Leu Ala Cys Ile Ser Met Asp Arg Tyr

130 135 140

Leu Ala Ile Val His Ala Thr Ser Thr Leu Ile Gln Lys Arg His Leu

145 150 155 160

Val Lys Phe Val Cys Ile Ala Met Trp Leu Leu Ser Val Ile Leu Ala

165 170 175

Leu Pro Ile Leu Ile Leu Arg Asn Pro Val Lys Val Asn Leu Ser Thr

180 185 190

Leu Val Cys Tyr Glu Asp Val Gly Asn Asn Thr Ser Arg Leu Arg Val

195 200 205

Val Leu Arg Ile Leu Pro Gln Thr Phe Gly Phe Leu Val Pro Leu Leu

210 215 220

Ile Met Leu Phe Cys Tyr Gly Phe Thr Leu Arg Thr Leu Phe Lys Ala

225 230 235 240

His Met Gly Gln Lys His Arg Ala Met Arg Val Ile Phe Ala Val Val

245 250 255

Leu Val Phe Leu Leu Cys Trp Leu Pro Tyr Asn Leu Val Leu Phe Thr

260 265 270

Asp Thr Leu Met Arg Thr Lys Leu Ile Lys Glu Thr Cys Glu Arg Arg

275 280 285

Asp Asp Ile Asp Lys Ala Leu Asn Ala Thr Glu Ile Leu Gly Phe Leu

290 295 300

His Ser Cys Leu Asn Pro Ile Ile Tyr Ala Phe Ile Gly Gln Lys Phe

305 310 315 320

Arg His Gly Leu Leu Lys Ile Met Ala Thr Tyr Gly Leu Val Ser Lys

325 330 335

Glu Phe Leu Ala Lys Glu Gly Arg Pro Ser Phe Val Ser Ser Ser Ser

340 345 350

Ala Asn Thr Ser Thr Thr Leu

355

<210> 127

<211> 355

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="cynomolgus CXCR2"

<400> 127

Met Gln Ser Phe Asn Phe Glu Asp Phe Trp Glu Asn Glu Asp Phe Ser

1 5 10 15

Asn Tyr Ser Tyr Ser Ser Asp Leu Pro Pro Ser Leu Pro Asp Val Ala

20 25 30

Pro Cys Arg Pro Glu Ser Leu Glu Ile Asn Lys Tyr Phe Val Val Ile

35 40 45

Ile Tyr Ala Leu Val Phe Leu Leu Ser Leu Leu Gly Asn Ser Leu Val

50 55 60

Met Leu Val Ile Leu His Ser Arg Val Gly Arg Ser Ile Thr Asp Val

65 70 75 80

Tyr Leu Leu Asn Leu Ala Met Ala Asp Leu Leu Phe Ala Leu Thr Leu

85 90 95

Pro Ile Trp Ala Ala Ala Lys Val Asn Gly Trp Ile Phe Gly Thr Phe

100 105 110

Leu Cys Lys Val Val Ser Leu Leu Lys Glu Val Asn Phe Tyr Ser Gly

115 120 125

Ile Leu Leu Leu Ala Cys Ile Ser Val Asp Arg Tyr Leu Ala Ile Val

130 135 140

His Ala Thr Arg Thr Leu Thr Gln Lys Arg Tyr Leu Val Lys Phe Val

145 150 155 160

Cys Leu Ser Ile Trp Ser Leu Ser Leu Leu Leu Ala Leu Pro Val Leu

165 170 175

Leu Phe Arg Arg Thr Val Tyr Leu Thr Tyr Ile Ser Pro Val Cys Tyr

180 185 190

Glu Asp Met Gly Asn Asn Thr Ala Lys Trp Arg Met Val Leu Arg Ile

195 200 205

Leu Pro Gln Thr Phe Gly Phe Ile Leu Pro Leu Leu Ile Met Leu Phe

210 215 220

Cys Tyr Gly Phe Thr Leu Arg Thr Leu Phe Lys Ala His Met Gly Gln

225 230 235 240

Lys His Arg Ala Met Arg Val Ile Phe Ala Val Val Leu Ile Phe Leu

245 250 255

Leu Cys Trp Leu Pro Tyr His Leu Val Leu Leu Ala Asp Thr Leu Met

260 265 270

Arg Thr Arg Leu Ile Asn Glu Thr Cys Gln Arg Arg Asn Asn Ile Asp

275 280 285

Gln Ala Leu Asp Ala Thr Glu Ile Leu Gly Ile Leu His Ser Cys Leu

290 295 300

Asn Pro Leu Ile Tyr Ala Phe Ile Gly Gln Lys Phe Arg His Gly Leu

305 310 315 320

Leu Lys Ile Leu Ala Thr His Gly Leu Ile Ser Lys Asp Ser Leu Pro

325 330 335

Lys Asp Ser Arg Pro Ser Phe Val Gly Ser Ser Ser Gly His Thr Ser

340 345 350

Thr Thr Leu

355

<210> 128

<211> 368

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="human CXCR3"

<400> 128

Met Val Leu Glu Val Ser Asp His Gln Val Leu Asn Asp Ala Glu Val

1 5 10 15

Ala Ala Leu Leu Glu Asn Phe Ser Ser Ser Tyr Asp Tyr Gly Glu Asn

20 25 30

Glu Ser Asp Ser Cys Cys Thr Ser Pro Pro Cys Pro Gln Asp Phe Ser

35 40 45

Leu Asn Phe Asp Arg Ala Phe Leu Pro Ala Leu Tyr Ser Leu Leu Phe

50 55 60

Leu Leu Gly Leu Leu Gly Asn Gly Ala Val Ala Ala Val Leu Leu Ser

65 70 75 80

Arg Arg Thr Ala Leu Ser Ser Thr Asp Thr Phe Leu Leu His Leu Ala

85 90 95

Val Ala Asp Thr Leu Leu Val Leu Thr Leu Pro Leu Trp Ala Val Asp

100 105 110

Ala Ala Val Gln Trp Val Phe Gly Ser Gly Leu Cys Lys Val Ala Gly

115 120 125

Ala Leu Phe Asn Ile Asn Phe Tyr Ala Gly Ala Leu Leu Leu Ala Cys

130 135 140

Ile Ser Phe Asp Arg Tyr Leu Asn Ile Val His Ala Thr Gln Leu Tyr

145 150 155 160

Arg Arg Gly Pro Pro Ala Arg Val Thr Leu Thr Cys Leu Ala Val Trp

165 170 175

Gly Leu Cys Leu Leu Phe Ala Leu Pro Asp Phe Ile Phe Leu Ser Ala

180 185 190

His His Asp Glu Arg Leu Asn Ala Thr His Cys Gln Tyr Asn Phe Pro

195 200 205

Gln Val Gly Arg Thr Ala Leu Arg Val Leu Gln Leu Val Ala Gly Phe

210 215 220

Leu Leu Pro Leu Leu Val Met Ala Tyr Cys Tyr Ala His Ile Leu Ala

225 230 235 240

Val Leu Leu Val Ser Arg Gly Gln Arg Arg Leu Arg Ala Met Arg Leu

245 250 255

Val Val Val Val Val Val Ala Phe Ala Leu Cys Trp Thr Pro Tyr His

260 265 270

Leu Val Val Leu Val Asp Ile Leu Met Asp Leu Gly Ala Leu Ala Arg

275 280 285

Asn Cys Gly Arg Glu Ser Arg Val Asp Val Ala Lys Ser Val Thr Ser

290 295 300

Gly Leu Gly Tyr Met His Cys Cys Leu Asn Pro Leu Leu Tyr Ala Phe

305 310 315 320

Val Gly Val Lys Phe Arg Glu Arg Met Trp Met Leu Leu Leu Arg Leu

325 330 335

Gly Cys Pro Asn Gln Arg Gly Leu Gln Arg Gln Pro Ser Ser Ser Arg

340 345 350

Arg Asp Ser Ser Trp Ser Glu Thr Ser Glu Ala Ser Tyr Ser Gly Leu

355 360 365

<210> 129

<211> 352

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="human CXCR4"

<400> 129

Met Glu Gly Ile Ser Ile Tyr Thr Ser Asp Asn Tyr Thr Glu Glu Met

1 5 10 15

Gly Ser Gly Asp Tyr Asp Ser Met Lys Glu Pro Cys Phe Arg Glu Glu

20 25 30

Asn Ala Asn Phe Asn Lys Ile Phe Leu Pro Thr Ile Tyr Ser Ile Ile

35 40 45

Phe Leu Thr Gly Ile Val Gly Asn Gly Leu Val Ile Leu Val Met Gly

50 55 60

Tyr Gln Lys Lys Leu Arg Ser Met Thr Asp Lys Tyr Arg Leu His Leu

65 70 75 80

Ser Val Ala Asp Leu Leu Phe Val Ile Thr Leu Pro Phe Trp Ala Val

85 90 95

Asp Ala Val Ala Asn Trp Tyr Phe Gly Asn Phe Leu Cys Lys Ala Val

100 105 110

His Val Ile Tyr Thr Val Asn Leu Tyr Ser Ser Val Leu Ile Leu Ala

115 120 125

Phe Ile Ser Leu Asp Arg Tyr Leu Ala Ile Val His Ala Thr Asn Ser

130 135 140

Gln Arg Pro Arg Lys Leu Leu Ala Glu Lys Val Val Tyr Val Gly Val

145 150 155 160

Trp Ile Pro Ala Leu Leu Leu Thr Ile Pro Asp Phe Ile Phe Ala Asn

165 170 175

Val Ser Glu Ala Asp Asp Arg Tyr Ile Cys Asp Arg Phe Tyr Pro Asn

180 185 190

Asp Leu Trp Val Val Val Phe Gln Phe Gln His Ile Met Val Gly Leu

195 200 205

Ile Leu Pro Gly Ile Val Ile Leu Ser Cys Tyr Cys Ile Ile Ile Ser

210 215 220

Lys Leu Ser His Ser Lys Gly His Gln Lys Arg Lys Ala Leu Lys Thr

225 230 235 240

Thr Val Ile Leu Ile Leu Ala Phe Phe Ala Cys Trp Leu Pro Tyr Tyr

245 250 255

Ile Gly Ile Ser Ile Asp Ser Phe Ile Leu Leu Glu Ile Ile Lys Gln

260 265 270

Gly Cys Glu Phe Glu Asn Thr Val His Lys Trp Ile Ser Ile Thr Glu

275 280 285

Ala Leu Ala Phe Phe His Cys Cys Leu Asn Pro Ile Leu Tyr Ala Phe

290 295 300

Leu Gly Ala Lys Phe Lys Thr Ser Ala Gln His Ala Leu Thr Ser Val

305 310 315 320

Ser Arg Gly Ser Ser Leu Lys Ile Leu Ser Lys Gly Lys Arg Gly Gly

325 330 335

His Ser Ser Val Ser Thr Glu Ser Glu Ser Ser Ser Phe His Ser Ser

340 345 350

<210> 130

<211> 372

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="human CXCR5"

<400> 130

Met Asn Tyr Pro Leu Thr Leu Glu Met Asp Leu Glu Asn Leu Glu Asp

1 5 10 15

Leu Phe Trp Glu Leu Asp Arg Leu Asp Asn Tyr Asn Asp Thr Ser Leu

20 25 30

Val Glu Asn His Leu Cys Pro Ala Thr Glu Gly Pro Leu Met Ala Ser

35 40 45

Phe Lys Ala Val Phe Val Pro Val Ala Tyr Ser Leu Ile Phe Leu Leu

50 55 60

Gly Val Ile Gly Asn Val Leu Val Leu Val Ile Leu Glu Arg His Arg

65 70 75 80

Gln Thr Arg Ser Ser Thr Glu Thr Phe Leu Phe His Leu Ala Val Ala

85 90 95

Asp Leu Leu Leu Val Phe Ile Leu Pro Phe Ala Val Ala Glu Gly Ser

100 105 110

Val Gly Trp Val Leu Gly Thr Phe Leu Cys Lys Thr Val Ile Ala Leu

115 120 125

His Lys Val Asn Phe Tyr Cys Ser Ser Leu Leu Leu Ala Cys Ile Ala

130 135 140

Val Asp Arg Tyr Leu Ala Ile Val His Ala Val His Ala Tyr Arg His

145 150 155 160

Arg Arg Leu Leu Ser Ile His Ile Thr Cys Gly Thr Ile Trp Leu Val

165 170 175

Gly Phe Leu Leu Ala Leu Pro Glu Ile Leu Phe Ala Lys Val Ser Gln

180 185 190

Gly His His Asn Asn Ser Leu Pro Arg Cys Thr Phe Ser Gln Glu Asn

195 200 205

Gln Ala Glu Thr His Ala Trp Phe Thr Ser Arg Phe Leu Tyr His Val

210 215 220

Ala Gly Phe Leu Leu Pro Met Leu Val Met Gly Trp Cys Tyr Val Gly

225 230 235 240

Val Val His Arg Leu Arg Gln Ala Gln Arg Arg Pro Gln Arg Gln Lys

245 250 255

Ala Val Arg Val Ala Ile Leu Val Thr Ser Ile Phe Phe Leu Cys Trp

260 265 270

Ser Pro Tyr His Ile Val Ile Phe Leu Asp Thr Leu Ala Arg Leu Lys

275 280 285

Ala Val Asp Asn Thr Cys Lys Leu Asn Gly Ser Leu Pro Val Ala Ile

290 295 300

Thr Met Cys Glu Phe Leu Gly Leu Ala His Cys Cys Leu Asn Pro Met

305 310 315 320

Leu Tyr Thr Phe Ala Gly Val Lys Phe Arg Ser Asp Leu Ser Arg Leu

325 330 335

Leu Thr Lys Leu Gly Cys Thr Gly Pro Ala Ser Leu Cys Gln Leu Phe

340 345 350

Pro Ser Trp Arg Arg Ser Ser Leu Ser Glu Ser Glu Asn Ala Thr Ser

355 360 365

Leu Thr Thr Phe

370

<210> 131

<211> 342

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="human CXCR6"

<400> 131

Met Ala Glu His Asp Tyr His Glu Asp Tyr Gly Phe Ser Ser Phe Asn

1 5 10 15

Asp Ser Ser Gln Glu Glu His Gln Asp Phe Leu Gln Phe Ser Lys Val

20 25 30

Phe Leu Pro Cys Met Tyr Leu Val Val Phe Val Cys Gly Leu Val Gly

35 40 45

Asn Ser Leu Val Leu Val Ile Ser Ile Phe Tyr His Lys Leu Gln Ser

50 55 60

Leu Thr Asp Val Phe Leu Val Asn Leu Pro Leu Ala Asp Leu Val Phe

65 70 75 80

Val Cys Thr Leu Pro Phe Trp Ala Tyr Ala Gly Ile His Glu Trp Val

85 90 95

Phe Gly Gln Val Met Cys Lys Ser Leu Leu Gly Ile Tyr Thr Ile Asn

100 105 110

Phe Tyr Thr Ser Met Leu Ile Leu Thr Cys Ile Thr Val Asp Arg Phe

115 120 125

Ile Val Val Val Lys Ala Thr Lys Ala Tyr Asn Gln Gln Ala Lys Arg

130 135 140

Met Thr Trp Gly Lys Val Thr Ser Leu Leu Ile Trp Val Ile Ser Leu

145 150 155 160

Leu Val Ser Leu Pro Gln Ile Ile Tyr Gly Asn Val Phe Asn Leu Asp

165 170 175

Lys Leu Ile Cys Gly Tyr His Asp Glu Ala Ile Ser Thr Val Val Leu

180 185 190

Ala Thr Gln Met Thr Leu Gly Phe Phe Leu Pro Leu Leu Thr Met Ile

195 200 205

Val Cys Tyr Ser Val Ile Ile Lys Thr Leu Leu His Ala Gly Gly Phe

210 215 220

Gln Lys His Arg Ser Leu Lys Ile Ile Phe Leu Val Met Ala Val Phe

225 230 235 240

Leu Leu Thr Gln Met Pro Phe Asn Leu Met Lys Phe Ile Arg Ser Thr

245 250 255

His Trp Glu Tyr Tyr Ala Met Thr Ser Phe His Tyr Thr Ile Met Val

260 265 270

Thr Glu Ala Ile Ala Tyr Leu Arg Ala Cys Leu Asn Pro Val Leu Tyr

275 280 285

Ala Phe Val Ser Leu Lys Phe Arg Lys Asn Phe Trp Lys Leu Val Lys

290 295 300

Asp Ile Gly Cys Leu Pro Tyr Leu Gly Val Ser His Gln Trp Lys Ser

305 310 315 320

Ser Glu Asp Asn Ser Lys Thr Phe Ser Ala Ser His Asn Val Glu Ala

325 330 335

Thr Ser Met Phe Gln Leu

340

<210> 132

<211> 362

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="human CXCR7"

<400> 132

Met Asp Leu His Leu Phe Asp Tyr Ser Glu Pro Gly Asn Phe Ser Asp

1 5 10 15

Ile Ser Trp Pro Cys Asn Ser Ser Asp Cys Ile Val Val Asp Thr Val

20 25 30

Met Cys Pro Asn Met Pro Asn Lys Ser Val Leu Leu Tyr Thr Leu Ser

35 40 45

Phe Ile Tyr Ile Phe Ile Phe Val Ile Gly Met Ile Ala Asn Ser Val

50 55 60

Val Val Trp Val Asn Ile Gln Ala Lys Thr Thr Gly Tyr Asp Thr His

65 70 75 80

Cys Tyr Ile Leu Asn Leu Ala Ile Ala Asp Leu Trp Val Val Leu Thr

85 90 95

Ile Pro Val Trp Val Val Ser Leu Val Gln His Asn Gln Trp Pro Met

100 105 110

Gly Glu Leu Thr Cys Lys Val Thr His Leu Ile Phe Ser Ile Asn Leu

115 120 125

Phe Gly Ser Ile Phe Phe Leu Thr Cys Met Ser Val Asp Arg Tyr Leu

130 135 140

Ser Ile Thr Tyr Phe Thr Asn Thr Pro Ser Ser Arg Lys Lys Met Val

145 150 155 160

Arg Arg Val Val Cys Ile Leu Val Trp Leu Leu Ala Phe Cys Val Ser

165 170 175

Leu Pro Asp Thr Tyr Tyr Leu Lys Thr Val Thr Ser Ala Ser Asn Asn

180 185 190

Glu Thr Tyr Cys Arg Ser Phe Tyr Pro Glu His Ser Ile Lys Glu Trp

195 200 205

Leu Ile Gly Met Glu Leu Val Ser Val Val Leu Gly Phe Ala Val Pro

210 215 220

Phe Ser Ile Ile Ala Val Phe Tyr Phe Leu Leu Ala Arg Ala Ile Ser

225 230 235 240

Ala Ser Ser Asp Gln Glu Lys His Ser Ser Arg Lys Ile Ile Phe Ser

245 250 255

Tyr Val Val Val Phe Leu Val Cys Trp Leu Pro Tyr His Val Ala Val

260 265 270

Leu Leu Asp Ile Phe Ser Ile Leu His Tyr Ile Pro Phe Thr Cys Arg

275 280 285

Leu Glu His Ala Leu Phe Thr Ala Leu His Val Thr Gln Cys Leu Ser

290 295 300

Leu Val His Cys Cys Val Asn Pro Val Leu Tyr Ser Phe Ile Asn Arg

305 310 315 320

Asn Tyr Arg Tyr Glu Leu Met Lys Ala Phe Ile Phe Lys Tyr Ser Ala

325 330 335

Lys Thr Gly Leu Thr Lys Leu Ile Asp Ala Ser Arg Val Ser Glu Thr

340 345 350

Glu Tyr Ser Ala Leu Glu Gln Ser Thr Lys

355 360

<210> 133

<211> 350

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="Human CXCR1"

<400> 133

Met Ser Asn Ile Thr Asp Pro Gln Met Trp Asp Phe Asp Asp Leu Asn

1 5 10 15

Phe Thr Gly Met Pro Pro Ala Asp Glu Asp Tyr Ser Pro Cys Met Leu

20 25 30

Glu Thr Glu Thr Leu Asn Lys Tyr Val Val Ile Ile Ala Tyr Ala Leu

35 40 45

Val Phe Leu Leu Ser Leu Leu Gly Asn Ser Leu Val Met Leu Val Ile

50 55 60

Leu Tyr Ser Arg Val Gly Arg Ser Val Thr Asp Val Tyr Leu Leu Asn

65 70 75 80

Leu Ala Leu Ala Asp Leu Leu Phe Ala Leu Thr Leu Pro Ile Trp Ala

85 90 95

Ala Ser Lys Val Asn Gly Trp Ile Phe Gly Thr Phe Leu Cys Lys Val

100 105 110

Val Ser Leu Leu Lys Glu Val Asn Phe Tyr Ser Gly Ile Leu Leu Leu

115 120 125

Ala Cys Ile Ser Val Asp Arg Tyr Leu Ala Ile Val His Ala Thr Arg

130 135 140

Thr Leu Thr Gln Lys Arg His Leu Val Lys Phe Val Cys Leu Gly Cys

145 150 155 160

Trp Gly Leu Ser Met Asn Leu Ser Leu Pro Phe Phe Leu Phe Arg Gln

165 170 175

Ala Tyr His Pro Asn Asn Ser Ser Pro Val Cys Tyr Glu Val Leu Gly

180 185 190

Asn Asp Thr Ala Lys Trp Arg Met Val Leu Arg Ile Leu Pro His Thr

195 200 205

Phe Gly Phe Ile Val Pro Leu Phe Val Met Leu Phe Cys Tyr Gly Phe

210 215 220

Thr Leu Arg Thr Leu Phe Lys Ala His Met Gly Gln Lys His Arg Ala

225 230 235 240

Met Arg Val Ile Phe Ala Val Val Leu Ile Phe Leu Leu Cys Trp Leu

245 250 255

Pro Tyr Asn Leu Val Leu Leu Ala Asp Thr Leu Met Arg Thr Gln Val

260 265 270

Ile Gln Glu Ser Cys Glu Arg Arg Asn Asn Ile Gly Arg Ala Leu Asp

275 280 285

Ala Thr Glu Ile Leu Gly Phe Leu His Ser Cys Leu Asn Pro Ile Ile

290 295 300

Tyr Ala Phe Ile Gly Gln Asn Phe Arg His Gly Phe Leu Lys Ile Leu

305 310 315 320

Ala Met His Gly Leu Val Ser Lys Glu Phe Leu Ala Arg His Arg Val

325 330 335

Thr Ser Tyr Thr Ser Ser Ser Val Asn Val Ser Ser Asn Leu

340 345 350

<210> 134

<211> 106

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="Human Lambda light chain constant region"

<400> 134

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro

35 40 45

Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

100 105

<210> 135

<211> 107

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="Human Kappa light chain constant region"

<400> 135

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 136

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S32D"

<400> 136

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Asp

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 137

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S35Q"

<400> 137

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 138

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S35D"

<400> 138

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 139

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH S35K"

<400> 139

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Lys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 140

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH A50S"

<400> 140

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 141

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH R55Q"

<400> 141

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Gln Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 142

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH R55D"

<400> 142

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Asp Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 143

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH R55H"

<400> 143

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly His Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 144

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96S"

<400> 144

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Ser Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 145

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96D"

<400> 145

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Asp Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 146

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96H"

<400> 146

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln His Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 147

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96L"

<400> 147

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Leu Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 148

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96W"

<400> 148

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Trp Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 149

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH M96Y"

<400> 149

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Tyr Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 150

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Y102Q"

<400> 150

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Gln Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 151

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Y102D"

<400> 151

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Asp Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 152

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V51D"

<400> 152

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 153

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V51Y"

<400> 153

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Tyr Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 154

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL R54D"

<400> 154

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Asp Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 155

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL Y91S"

<400> 155

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Ser Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 156

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL Y91H"

<400> 156

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser His Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 157

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N94H"

<400> 157

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly His

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 158

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aS"

<400> 158

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Ser Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 159

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL N95aW"

<400> 159

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Trp Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 160

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V97S"

<400> 160

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Ser Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 161

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VL V97D"

<400> 161

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Asp Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 162

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 102 M34Q_A40P_N56S_R75K_M96E"

<400> 162

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Glu Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 163

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 103 M34Q_A40P_R53S_R55G_N56S_R75K"

<400> 163

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 164

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 104 M34Q_A40P_R53S_R55G_N56S_R75K_M96K"

<400> 164

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Lys Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 165

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 105 M34Q_A40P_R53S_R55G_N56S_R75K_M96A"

<400> 165

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Ala Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 166

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 VH Variant 106 M34Q_A40P_R53S_R55G_N56S_R75K_M96E"

<400> 166

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Gln Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Glu Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 167

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 Consensus VH"

<220>

<221> VARIANT

<222> (32)..(32)

<223> /replace="Q" or "H" or "L" or "W" or "Y"

<220>

<221> VARIANT

<222> (33)..(33)

<223> /replace="A"

<220>

<221> VARIANT

<222> (34)..(34)

<223> /replace="Q" or "D" or "H" or "W"

<220>

<221> VARIANT

<222> (51)..(51)

<223> /replace="H"

<220>

<221> VARIANT

<222> (53)..(53)

<223> /replace="D"

<220>

<221> VARIANT

<222> (54)..(54)

<223> /replace="S" or "Q"

<220>

<221> VARIANT

<222> (55)..(55)

<223> /replace="D"

<220>

<221> VARIANT

<222> (57)..(57)

<223> /replace="S"

<220>

<221> VARIANT

<222> (98)..(98)

<223> /replace="K"

<220>

<221> VARIANT

<222> (100)..(100)

<223> /replace="A" or "Q" or "K"

<220>

<221> VARIANT

<222> (101)..(101)

<223> /replace="D"

<220>

<221> VARIANT

<222> (102)..(102)

<223> /replace="S" or "K"

<220>

<221> SITE

<222> (1)..(113)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 167

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 168

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 Consensus VL"

<220>

<221> VARIANT

<222> (52)..(52)

<223> /replace="D"

<220>

<221> VARIANT

<222> (54)..(54)

<223> /replace="D" or "S"

<220>

<221> VARIANT

<222> (55)..(55)

<223> /replace="A" or "D" or "H"

<220>

<221> VARIANT

<222> (56)..(56)

<223> /replace="Q"

<220>

<221> VARIANT

<222> (93)..(93)

<223> /replace="A"

<220>

<221> VARIANT

<222> (96)..(96)

<223> /replace="A" or "S" or "K" or "L" or "W" or "Y"

<220>

<221> VARIANT

<222> (98)..(98)

<223> /replace="Q" or "D" or "H" or "K" or "L" or "Y"

<220>

<221> VARIANT

<222> (101)..(101)

<223> /replace="A" or "K"

<220>

<221> SITE

<222> (1)..(111)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 168

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 169

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR1"

<220>

<221> VARIANT

<222> (2)..(2)

<223> /replace="Q" or "H" or "L" or "W" or "Y"

<220>

<221> VARIANT

<222> (3)..(3)

<223> /replace="A"

<220>

<221> VARIANT

<222> (4)..(4)

<223> /replace="Q" or "D" or "H" or "W"

<220>

<221> SITE

<222> (1)..(5)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 169

Ser Ser Thr Met Ser

15

<210> 170

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR2"

<220>

<221> VARIANT

<222> (2)..(2)

<223> /replace="H"

<220>

<221> VARIANT

<222> (4)..(4)

<223> /replace="D"

<220>

<221> VARIANT

<222> (5)..(5)

<223> /replace="S" or "Q"

<220>

<221> VARIANT

<222> (6)..(6)

<223> /replace="D"

<220>

<221> VARIANT

<222> (8)..(8)

<223> /replace="S"

<220>

<221> SITE

<222> (1)..(17)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 170

Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 171

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR3"

<220>

<221> VARIANT

<222> (2)..(2)

<223> /replace="A" or "Q" or "K"

<220>

<221> VARIANT

<222> (3)..(3)

<223> /replace="D"

<220>

<221> VARIANT

<222> (4)..(4)

<223> /replace="S" or "K"

<220>

<221> SITE

<222> (1)..(4)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 171

Gln Met Gly Tyr

1

<210> 172

<211> 14

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VL CDR1"

<400> 172

Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser

1 5 10

<210> 173

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VL CDR2"

<220>

<221> VARIANT

<222> (1)..(1)

<223> /replace="D"

<220>

<221> VARIANT

<222> (3)..(3)

<223> /replace="D" or "S"

<220>

<221> VARIANT

<222> (4)..(4)

<223> /replace="A" or "D" or "H"

<220>

<221> VARIANT

<222> (5)..(5)

<223> /replace="Q"

<220>

<221> SITE

<222> (1)..(7)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 173

Glu Val Asn Lys Arg Pro Ser

15

<210> 174

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VL CDR3"

<220>

<221> VARIANT

<222> (3)..(3)

<223> /replace="A"

<220>

<221> VARIANT

<222> (6)..(6)

<223> /replace="A" or "S" or "K" or "L" or "W" or "Y"

<220>

<221> VARIANT

<222> (8)..(8)

<223> /replace="Q" or "D" or "H" or "K" or "L" or "Y"

<220>

<221> VARIANT

<222> (11)..(11)

<223> /replace="A" or "K"

<220>

<221> SITE

<222> (1)..(11)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 174

Ser Ser Tyr Ala Gly Asn Asn Asn Phe Gly Val

1 5 10

<210> 175

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VHCDR1"

<400> 175

Ser Ser Thr Met Ser

15

<210> 176

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 S32Q VHCDR1"

<400> 176

Ser Gln Thr Met Ser

15

<210> 177

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 S32H VHCDR1"

<400> 177

Ser His Thr Met Ser

15

<210> 178

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 S32L VHCDR1"

<400> 178

Ser Leu Thr Met Ser

15

<210> 179

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 S32W VHCDR1"

<400> 179

Ser Trp Thr Met Ser

15

<210> 180

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 S32Y VHCDR1"

<400> 180

Ser Tyr Thr Met Ser

15

<210> 181

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 T33A VHCDR1"

<400> 181

Ser Ser Ala Met Ser

15

<210> 182

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M34Q VHCDR1"

<400> 182

Ser Ser Thr Gln Ser

15

<210> 183

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M34D VHCDR1"

<400> 183

Ser Ser Thr Asp Ser

15

<210> 184

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M34H VHCDR1"

<400> 184

Ser Ser Thr His Ser

15

<210> 185

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M34W VHCDR1"

<400> 185

Ser Ser Thr Trp Ser

15

<210> 186

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VHCDR2"

<400> 186

Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 187

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 I51H VHCDR2"

<400> 187

Ala His Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 188

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 G52aD VHCDR2"

<400> 188

Ala Ile Ser Asp Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 189

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 R53S VHCDR2"

<400> 189

Ala Ile Ser Gly Ser Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 190

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 R53Q VHCDR2"

<400> 190

Ala Ile Ser Gly Gln Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 191

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 G54D VHCDR2"

<400> 191

Ala Ile Ser Gly Arg Asp Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 192

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N56S VHCDR2"

<400> 192

Ala Ile Ser Gly Arg Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 193

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 103,104, 105 VHCDR2"

<400> 193

Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 194

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VHCDR3"

<400> 194

Gln Met Gly Tyr

1

<210> 195

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M96A VHCDR3"

<400> 195

Gln Ala Gly Tyr

1

<210> 196

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M96Q VHCDR3"

<400> 196

Gln Gln Gly Tyr

1

<210> 197

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 M96K VHCDR3"

<400> 197

Gln Lys Gly Tyr

1

<210> 198

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 G101D VHCDR3"

<400> 198

Gln Met Asp Tyr

1

<210> 199

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 Y102S VHCDR3"

<400> 199

Gln Met Gly Ser

1

<210> 200

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 Y102K VHCDR3"

<400> 200

Gln Met Gly Lys

1

<210> 201

<211> 14

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VLCDR1 and Consensus VL CDR1"

<400> 201

Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser

1 5 10

<210> 202

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VLCDR2"

<400> 202

Glu Val Asn Lys Arg Pro Ser

15

<210> 203

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 E50D VLCDR2"

<400> 203

Asp Val Asn Lys Arg Pro Ser

15

<210> 204

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N52D VLCDR2"

<400> 204

Glu Val Asp Lys Arg Pro Ser

15

<210> 205

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N52S VLCDR2"

<400> 205

Glu Val Ser Lys Arg Pro Ser

15

<210> 206

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 K53A VLCDR2"

<400> 206

Glu Val Asn Ala Arg Pro Ser

15

<210> 207

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 K53D VLCDR2"

<400> 207

Glu Val Asn Asp Arg Pro Ser

15

<210> 208

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 K53H VLCDR2"

<400> 208

Glu Val Asn His Arg Pro Ser

15

<210> 209

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 R54Q VLCDR2"

<400> 209

Glu Val Asn Lys Gln Pro Ser

15

<210> 210

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 VLCDR3"

<400> 210

Ser Ser Tyr Ala Gly Asn Asn Asn Phe Gly Val

1 5 10

<210> 211

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 Y91A VLCDR3"

<400> 211

Ser Ser Ala Ala Gly Asn Asn Asn Phe Gly Val

1 5 10

<210> 212

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94A VLCDR3"

<400> 212

Ser Ser Tyr Ala Gly Ala Asn Asn Phe Gly Val

1 5 10

<210> 213

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94S VLCDR3"

<400> 213

Ser Ser Tyr Ala Gly Ser Asn Asn Phe Gly Val

1 5 10

<210> 214

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94K VLCDR3"

<400> 214

Ser Ser Tyr Ala Gly Lys Asn Asn Phe Gly Val

1 5 10

<210> 215

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94L VLCDR3"

<400> 215

Ser Ser Tyr Ala Gly Leu Asn Asn Phe Gly Val

1 5 10

<210> 216

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94W VLCDR3"

<400> 216

Ser Ser Tyr Ala Gly Trp Asn Asn Phe Gly Val

1 5 10

<210> 217

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N94Y VLCDR3"

<400> 217

Ser Ser Tyr Ala Gly Tyr Asn Asn Phe Gly Val

1 5 10

<210> 218

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N95aQ VLCDR3"

<400> 218

Ser Ser Tyr Ala Gly Asn Asn Gln Phe Gly Val

1 5 10

<210> 219

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N95aD VLCDR3"

<400> 219

Ser Ser Tyr Ala Gly Asn Asn Asp Phe Gly Val

1 5 10

<210> 220

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N95aH VLCDR3"

<400> 220

Ser Ser Tyr Ala Gly Asn Asn His Phe Gly Val

1 5 10

<210> 221

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N95aK VLCDR3"

<400> 221

Ser Ser Tyr Ala Gly Asn Asn Lys Phe Gly Val

1 5 10

<210> 222

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 N95aL VLCDR3"

<400> 222

Ser Ser Tyr Ala Gly Asn Asn Leu Phe Gly Val

1 5 10

<210> 223

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="A48 N95aYVLCDR3"

<400> 223

Ser Ser Tyr Ala Gly Asn Asn Tyr Phe Gly Val

1 5 10

<210> 224

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 V97A VLCDR3"

<400> 224

Ser Ser Tyr Ala Gly Asn Asn Asn Phe Gly Ala

1 5 10

<210> 225

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="4A8 V97K VLCDR3"

<400> 225

Ser Ser Tyr Ala Gly Asn Asn Asn Phe Gly Lys

1 5 10

<210> 226

<211> 113

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 Consensus VH"

<220>

<221> VARIANT

<222> (34)..(34)

<223> /replace="Q"

<220>

<221> VARIANT

<222> (40)..(40)

<223> /replace="P"

<220>

<221> VARIANT

<222> (54)..(54)

<223> /replace="S"

<220>

<221> VARIANT

<222> (56)..(56)

<223> /replace="G"

<220>

<221> VARIANT

<222> (57)..(57)

<223> /replace="S"

<220>

<221> VARIANT

<222> (76)..(76)

<223> /replace="K"

<220>

<221> VARIANT

<222> (98)..(98)

<223> /replace="K"

<220>

<221> VARIANT

<222> (100)..(100)

<223> /replace="K" or "A"

<220>

<221> SITE

<222> (1)..(113)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 226

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser

20 25 30

Thr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ile Gln Met Gly Tyr Trp Gly Gln Gly Ile Leu Val Thr Val Ser

100 105 110

Ser

<210> 227

<211> 111

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polypeptide"

<220>

<221> source

<223> /note="4A8 Consensus VL"

<220>

<221> VARIANT

<222> (43)..(43)

<223> /replace="G"

<220>

<221> VARIANT

<222> (54)..(54)

<223> /replace="S"

<220>

<221> VARIANT

<222> (96)..(96)

<223> /replace="S"

<220>

<221> SITE

<222> (1)..(111)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 227

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Ile Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Asp Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asn

85 90 95

Asn Asn Phe Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 228

<211> 5

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR1"

<220>

<221> VARIANT

<222> (4)..(4)

<223> /replace="Q"

<220>

<221> SITE

<222> (1)..(5)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 228

Ser Ser Thr Met Ser

15

<210> 229

<211> 17

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR2"

<220>

<221> VARIANT

<222> (5)..(5)

<223> /replace="S"

<220>

<221> VARIANT

<222> (7)..(7)

<223> /replace="G"

<220>

<221> VARIANT

<222> (8)..(8)

<223> /replace="S"

<220>

<221> SITE

<222> (1)..(17)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 229

Ala Ile Ser Gly Arg Gly Arg Asn Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 230

<211> 4

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VH CDR3"

<220>

<221> VARIANT

<222> (2)..(2)

<223> /replace="K" or "A"

<220>

<221> SITE

<222> (1)..(4)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 230

Gln Met Gly Tyr

1

<210> 231

<211> 7

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VL CDR2"

<220>

<221> VARIANT

<222> (3)..(3)

<223> /replace="S"

<220>

<221> SITE

<222> (1)..(7)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 231

Glu Val Asn Lys Arg Pro Ser

15

<210> 232

<211> 11

<212>PRT

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

peptide"

<220>

<221> source

<223> /note="Consensus VL CDR3"

<220>

<221> VARIANT

<222> (6)..(6)

<223> /replace="S"

<220>

<221> SITE

<222> (1)..(11)

<223> /note="Variant residues given in the sequence have no

preference with respect to those in the annotations

for variant positions"

<400> 232

Ser Ser Tyr Ala Gly Asn Asn Asn Phe Gly Val

1 5 10

<210> 233

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="BKO-4A8 VH"

<400> 233

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacaa tgagctgggt ccgacaggcc

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggaa cacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagatg

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 234

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 1"

<400> 234

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacaggcc

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagatg

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 235

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 2"

<400> 235

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagatg

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 236

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 3"

<400> 236

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcaaaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagatg

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 237

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 4"

<400> 237

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 238

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 5"

<400> 238

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcaaaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 239

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 6"

<400> 239

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcaaaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc caagcagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 240

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 7"

<400> 240

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacagcct

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc caagcagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 241

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 8"

<400> 241

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacaggcc

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 242

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 9"

<400> 242

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacac agagctgggt ccgacaggcc

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggag tacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcaaaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc catccagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 243

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 10"

<400> 243

gaagtgcagc tgctggaatc tggcggagga ctggtgcagc ctggcggcag cctgagactg

60

tcttgtgccg ccagcggctt caccttcagc agcagcacaa tgagctgggt ccgacaggcc

120

cctggcaagg gactggaatg ggtgtccgcc atcagcggca gaggccggaa cacctactac

180

gccgacagcg tgaagggccg gttcaccatc agccgggaca acagcagaaa caccctgtac

240

ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtgc caagcagaag

300

ggctactggg gccagggcat tctcgtgaca gtgtcctca

339

<210> 244

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 101"

<400> 244

gaagttcagc tgcttgaatc tggcggagga ctggttcagc ctggcggatc tctgagactg

60

tcttgtgccg ccagcggctt cacctttagc agcagcacac agagctgggt ccgacagcct

120

cctggcaaag gactggaatg ggtgtccgcc atctctggca gaggcagaag cacctactac

180

gccgactctg tgaagggcag attcaccatc agccgggaca acagcaagaa caccctgtac

240

ctgcagatga acagcctgag agccgaggac accgccgtgt actattgtgc catccaggcc

300

ggctattggg gccagggaat actcgtgaca gtgtcctca

339

<210> 245

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 103"

<400> 245

gaagttcagc tgcttgaatc tggcggagga ctggttcagc ctggcggatc tctgagactg

60

tcttgtgccg ccagcggctt cacctttagc agcagcacac agagctgggt ccgacagcct

120

cctggcaaag gactggaatg ggtgtccgcc atctctggca gcggcggcag cacatattac

180

gccgattctg tgaagggcag attcaccatc agccgggaca acagcaagaa caccctgtac

240

ctgcagatga acagcctgag agccgaggac accgccgtgt actattgcgc catccagatg

300

ggctattggg gccagggaat cctcgtgaca gtgtcctca

339

<210> 246

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 104"

<400> 246

gaagttcagc tgcttgaatc tggcggagga ctggttcagc ctggcggatc tctgagactg

60

tcttgtgccg ccagcggctt cacctttagc agcagcacac agagctgggt ccgacagcct

120

cctggcaaag gactggaatg ggtgtccgcc atctctggca gcggcggcag cacatattac

180

gccgattctg tgaagggcag attcaccatc agccgggaca acagcaagaa caccctgtac

240

ctgcagatga acagcctgag agccgaggac accgccgtgt actattgcgc catccagaaa

300

ggctattggg gccagggcat cctcgtgaca gtgtcctca

339

<210> 247

<211> 339

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VH Variant 105"

<400> 247

gaagttcagc tgcttgaatc tggcggagga ctggttcagc ctggcggatc tctgagactg

60

tcttgtgccg ccagcggctt cacctttagc agcagcacac agagctgggt ccgacagcct

120

cctggcaaag gactggaatg ggtgtccgcc atctctggca gcggcggcag cacatattac

180

gccgattctg tgaagggcag attcaccatc agccgggaca acagcaagaa caccctgtac

240

ctgcagatga acagcctgag agccgaggac accgccgtgt actattgtgc catccaggcc

300

ggctattggg gccagggaat actcgtgaca gtgtcctca

339

<210> 248

<211> 987

<212> DNA

<213> Homo sapiens

<220>

<221> source

<223> /note="Human IgG1"

<400> 248

gctagcacca agggacccag cgtgttcccc ctggccccca gcagcaagag cacatctggc

60

ggaacagccg ccctgggctg cctggtgaaa gactacttcc ccgagcccgt gaccgtgagc

120

tggaacagcg gagccctgac cagcggcgtg cacacctttc cagccgtgct gcagagcagc

180

ggcctgtaca gcctgagcag cgtggtgaca gtgccctcta gcagcctggg cacccagacc

240

tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaaaaa ggtggaaccc

300

aagagctgcg acaagaccca cacctgtccc ccctgccctg cccctgaact gctgggcgga

360

ccctccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc

420

gaagtgacct gcgtggtggt ggacgtgtcc cacgaggacc ctgaagtgaa gttcaattgg

480

tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ccagagagga acagtacaac

540

agcacctacc gggtggtgtc cgtgctgacc gtgctgcacc aggactggct gaacggcaaa

600

gagtacaagt gcaaggtgtc caacaaggcc ctgcctgctc ccatcgagaa aaccatcagc

660

aaggccaagg gccagccccg cgagcctcag gtgtacacac tgccccccag ccgggacgag

720

ctgaccaaga accaggtgtc cctgacctgt ctggtgaaag gcttctaccc cagcgatatc

780

gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg

840

ctggacagcg acggctcatt cttcctgtac agcaagctga ccgtggacaa gagccggtgg

900

cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc

960

cagaagtccc tgagcctgag ccccggc

987

<210> 249

<211> 987

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="Human IgG1"

<400> 249

gctagcacca agggacccag cgtgttcccc ctggccccca gcagcaagag cacatctggc

60

ggaacagccg ccctgggctg cctggtgaaa gactacttcc ccgagcccgt gaccgtgagc

120

tggaacagcg gagccctgac cagcggcgtg cacacctttc cagccgtgct gcagagcagc

180

ggcctgtaca gcctgagcag cgtggtgaca gtgccctcta gcagcctggg cacccagacc

240

tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaaaaa ggtggaaccc

300

aagagctgcg acaagaccca cacctgtccc ccctgccctg cccctgaact ggctggcgct

360

ccctccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc

420

gaagtgacct gcgtggtggt ggacgtgtcc cacgaggacc ctgaagtgaa gttcaattgg

480

tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ccagagagga acagtacaac

540

agcacctacc gggtggtgtc cgtgctgacc gtgctgcacc aggactggct gaacggcaaa

600

gagtacaagt gcaaggtgtc caacaaggcc ctgcctgctc ccatcgagaa aaccatcagc

660

aaggccaagg gccagccccg cgagcctcag gtgtacacac tgccccccag ccgggacgag

720

ctgaccaaga accaggtgtc cctgacctgt ctggtgaaag gcttctaccc cagcgatatc

780

gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg

840

ctggacagcg acggctcatt cttcctgtac agcaagctga ccgtggacaa gagccggtgg

900

cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc

960

cagaagtccc tgagcctgag ccccggc

987

<210> 250

<211> 978

<212> DNA

<213> Homo sapiens

<220>

<221> source

<223> /note="Human IgG4"

<400> 250

gctagcacca agggccccag cgtgttcccc ctggcccctt gtagcagaag caccagcgag

60

agcacagccg ccctgggctg cctggtgaaa gactacttcc ccgagcccgt caccgtgtcc

120

tggaacagcg gagccctgac cagcggcgtg cacacctttc cagccgtgct gcagagcagc

180

ggcctgtaca gcctgagcag cgtggtgaca gtgccctcca gcagcctggg caccaagacc

240

tacacctgta acgtggacca caagcccagc aacaccaagg tggacaagcg ggtggaatct

300

aagtacggcc caccctgccc cccctgccct gcccctgaat ttctgggcgg accctccgtg

360

ttcctgttcc ccccaaagcc caaggacacc ctgatgatca gccggacccc cgaagtgacc

420

tgcgtggtgg tggacgtgtc cccaggaagat cccgaggtcc agttcaattg gtacgtggac

480

ggcgtggaag tgcacaacgc caagaccaag cccagagagg aacagttcaa cagcacctac

540

cgggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agagtacaag

600

tgcaaagtct ccaacaaggg cctgcccagc tccatcgaga aaaccatcag caaggccaag

660

ggccagcccc gcgagcctca ggtgtacaca ctgcccccca gccaggaaga gatgaccaag

720

aaccaggtgt ccctgacctg tctggtgaaa ggcttctacc ccagcgatat cgccgtggaa

780

tggggagagca acggccagcc cgagaacaac tacaagacca ccccccctgt gctggacagc

840

gacggcagct tcttcctgta ctcccggctg accgtggaca agagccggtg gcaggaaggc

900

aacgtcttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc

960

ctgagcctga gcctggggc

978

<210> 251

<211> 978

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="Human IgG4"

<400> 251

gctagcacca agggccccag cgtgttcccc ctggcccctt gtagcagaag caccagcgag

60

agcacagccg ccctgggctg cctggtgaaa gactacttcc ccgagcccgt caccgtgtcc

120

tggaacagcg gagccctgac cagcggcgtg cacacctttc cagccgtgct gcagagcagc

180

ggcctgtaca gcctgagcag cgtggtgaca gtgccctcca gcagcctggg caccaagacc

240

tacacctgta acgtggacca caagcccagc aacaccaagg tggacaagcg ggtggaatct

300

aagtacggcc caccctgccc cccctgccct gcccctgaat ttctgggcgg accctccgtg

360

ttcctgttcc ccccaaagcc caaggacacc ctgtatatca ctcggggagcc cgaagtgacc

420

tgcgtggtgg tggacgtgtc cccaggaagat cccgaggtcc agttcaattg gtacgtggac

480

ggcgtggaag tgcacaacgc caagaccaag cccagagagg aacagttcaa cagcacctac

540

cgggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agagtacaag

600

tgcaaagtct ccaacaaggg cctgcccagc tccatcgaga aaaccatcag caaggccaag

660

ggccagcccc gcgagcctca ggtgtacaca ctgcccccca gccaggaaga gatgaccaag

720

aaccaggtgt ccctgacctg tctggtgaaa ggcttctacc ccagcgatat cgccgtggaa

780

tggggagagca acggccagcc cgagaacaac tacaagacca ccccccctgt gctggacagc

840

gacggcagct tcttcctgta ctcccggctg accgtggaca agagccggtg gcaggaaggc

900

aacgtcttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc

960

ctgagcctga gcctggggc

978

<210> 252

<211> 975

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="Human IgG2"

<400> 252

gctagcacca agggccccag cgtgttccct ctggcccctt gtagcagaag caccagcgag

60

tctacagccg ccctgggctg cctcgtgaag gactactttc ccgagcccgt caccgtgtcc

120

tggaactctg gcgctctgac aagcggcgtg cacacctttc cagccgtgct gcagagcagc

180

ggcctgtact ctctgagcag cgtcgtgacc gtgcccagca gcaatttcgg cacccagacc

240

tacacctgta acgtggacca caagcccagc aacaccaagg tggacaagac cgtggaacgg

300

aagtgctgcg tggaatgccc cccttgtcct gcccctccag tggctggccc ttccgtgttc

360

ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc

420

gtggtggtgg atgtgtccca cgaggacccc gaggtgcagt tcaattggta cgtggacggc

480

gtggaagtgc acaacgccaa gaccaagccc agagaggaac agttcaacag caccttccgg

540

gtggtgtccg tgctgaccgt ggtgcatcag gactggctga acggcaaaga gtacaagtgc

600

aaggtgtcca acaagggcct gcccagctcc atcgagaaaa ccatcagcaa gaccaaaggc

660

cagccccgcg agccccaggt gtacacactg cctccaagcc gggaagagat gaccaagaat

720

caggtgtccc tgacctgtct cgtgaaaggc ttctacccct ccgatatcgc cgtggaatgg

780

gagagcaacg gccagcccga gaacaactac aagaccaccc ccccatgct ggacagcgac

840

ggctcattct tcctgtacag caagctgaca gtggacaagt cccggtggca gcagggcaac

900

gtgttcagct gcagcgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg

960

agcctgagcc ctggc

975

<210> 253

<211> 333

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="BKO-4A8 VL"

<400> 253

cagtctgctc tgacacagcc tcctagcgcc tctggctctc ctggccagag cgtgaccatc

60

agctgtatcg gcaccagcag cgacgtgggc ggctacaact acgtgtcctg gtatcagcag

120

caccccgaca aggcccccaa gctgatgatc tacgaagtga acaagcggcc cagcggcgtg

180

cccgatagat tcagcggcag caagagcggc aacaccgcca gcctcacagt gtctggactg

240

caggccgagg acgaggccga ctactactgt agcagctacg ccggcaacaa caacttcggc

300

gtgttcggcg gaggcaccaa gctgacagtc cta

333

<210> 254

<211> 333

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VL variant b"

<400> 254

cagtctgctc tgacacagcc tcctagcgcc tctggctctc ctggccagag cgtgaccatc

60

agctgtatcg gcaccagcag cgacgtgggc ggctacaact acgtgtcctg gtatcagcag

120

caccccgaca aggcccccaa gctgatgatc tacgaagtgt ccaagcggcc cagcggcgtg

180

cccgatagat tcagcggcag caagagcggc aacaccgcca gcctcacagt gtctggactg

240

caggccgagg acgaggccga ctactactgt agcagctacg ccggcagcaa caacttcggc

300

gtgttcggcg gaggcaccaa gctgacagtc cta

333

<210> 255

<211> 333

<212> DNA

<213> Artificial Sequence

<220>

<221> source

<223> /note="Description of Artificial Sequence: Synthetic

polynucleotide"

<220>

<221> source

<223> /note="Completely synthesized"

<220>

<221> source

<223> /note="4A8 VL variant c"

<400> 255

cagtctgctc tgacacagcc tcctagcgcc tctggctctc ctggccagag cgtgaccatc

60

agctgtatcg gcaccagcag cgacgtgggc ggctacaact acgtgtcctg gtatcagcag

120

caccccggta aggcccccaa gctgatgatc tacgaagtgt ccaagcggcc cagcggcgtg

180

cccgatagat tcagcggcag caagagcggc aacaccgcca gcctcacagt gtctggactg

240

caggccgagg acgaggccga ctactactgt agcagctacg ccggcagcaa caacttcggc

300

gtgttcggcg gaggcaccaa gctgacagtc cta

333

<210> 256

<211> 318

<212> DNA

<213> Homo sapiens

<220>

<221> source

<223> /note="lambda constant light chain"

<400> 256

ggtcagccca aggccgctcc cagcgtgacc ctgttccccc caagcagcga ggaactgcag

60

gccaacaagg ccaccctggt gtgcctgatc agcgacttct accctggggc cgtgaccgtg

120

gcctggaagg ccgatagcag ccctgtgaag gccggcgtgg aaaccaccac cccctccaag

180

cagagcaaca acaaatacgc cgccagcagc tacctgtccc tgacccccga gcagtggaag

240

tcccaccggt cctacagctg ccaggtgaca cacgagggca gcaccgtgga aaagaccgtg

300

gccccccaccg agtgcagc

318

<---

Claims (53)

1. A human antibody molecule that immunospecifically binds to human CXCR2, the antibody molecule comprising: Heavy chain CDR1 having the amino acid sequence SEQ ID NO: 169, Heavy chain CDR2 having the amino acid sequence SEQ ID NO: 170, Heavy chain CDR3 having the amino acid sequence SEQ ID NO: 171, Light chain CDR1 having the amino acid sequence SEQ ID NO: 172, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 173, and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 174; or Heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 228, Heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 229, Heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 230, Light chain CDR1 having the amino acid sequence of SEQ ID NO: : 201, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 231, and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 232, and wherein the antibody molecule inhibits CXCL1-induced activation of CXCR2 or CXCL5-induced activation of CXCR2. 2. Human antibody molecule according to claim 1, wherein: the heavy chain variable region has the amino acid sequence of SEQ ID NO: 167, and the light chain variable region has the amino acid sequence of SEQ ID NO: 168; or the heavy chain variable region has the amino acid sequence of SEQ ID NO: 226 and the light chain variable region has the amino acid sequence of SEQ ID NO: 227. 3. Human antibody molecule according to claim 2, wherein: a) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 98 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; b) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 99, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; c) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 100, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; d) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 101 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; e) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 102 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; f) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 103 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; g) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 104 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; h) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 105 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; i) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 106 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; j) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 107 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; k) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; l) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 162, and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; m) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 163 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; n) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 164 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; o) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 165 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110; or p) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 166 and the light chain variable region has the amino acid sequence of SEQ ID NO: 109 or 110. 4. Human antibody molecule according to claim 3, wherein: a) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; b) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 162, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; c) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 163 and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; d) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 164 and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; e) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 165 and the light chain variable region has the amino acid sequence of SEQ ID NO: 110; or f) the heavy chain variable region has the amino acid sequence of SEQ ID NO: 166, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110. 5. The human antibody molecule according to claim 1, wherein the heavy chain CDR1 has the amino acid sequence SEQ ID NO: 182, the heavy chain CDR2 has the amino acid sequence SEQ ID NO: 192, the heavy chain CDR3 has the amino acid sequence SEQ ID NO: 195, the light CDR1 chain has the amino acid sequence of SEQ ID NO: 201, the light chain CDR2 has the amino acid sequence of SEQ ID NO: 205, and the light chain CDR3 has the amino acid sequence of SEQ ID NO: 213. 6. Human antibody molecule according to any one of paragraphs. 1-5, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 108, and the light chain variable region has the amino acid sequence of SEQ ID NO: 110. 7. Human antibody molecule according to any one of paragraphs. 1-6, wherein the antibody contains the human IgG1 heavy chain constant region. 8. The human antibody molecule according to claim 7, wherein the human IgG1 heavy chain constant region has the amino acid sequence of SEQ ID NO: 122 or 124. 9. Human antibody molecule according to any one of paragraphs. 1-6, wherein the antibody contains the human IgG2 heavy chain constant region. 10. The human antibody molecule according to claim 9, wherein the human IgG2 heavy chain constant region has the amino acid sequence SEQ ID NO: 120. 11. Human antibody molecule according to any one of paragraphs. 1-6, wherein the antibody contains the human IgG4 heavy chain constant region. 12. Human antibody molecule according to any one of paragraphs. 1-11, wherein the antibody molecule is a Fab fragment, an F(ab) ₂ fragment, or a single chain antibody. 13. Pharmaceutical composition for the treatment or prevention of respiratory tract neutrophilia or acute pneumonia, containing a therapeutically effective amount of a human antibody molecule according to any one of paragraphs. 1-12. 14. A nucleic acid molecule encoding a human antibody molecule according to any one of claims. 1-12. 15. An expression vector containing a nucleic acid molecule according to claim 14. 16. A cell transformed to express a human antibody molecule according to any one of claims. 1-12. 17. A method of treating or preventing airway neutrophilia or acute pneumonia in a subject, which includes administering to a subject a therapeutically effective amount of human antibody molecules according to any one of claims. 1-12 or a pharmaceutical composition according to claim 13 for the prevention or treatment of airway neutrophilia or acute pneumonia. 18. The method of claim 17, wherein the airway neutrophilia or acute pneumonia, or both, is chronic obstructive pulmonary disease, severe neutrophilic asthma, or both. 19. Human antibody molecule according to any one of paragraphs. 1-12 or a pharmaceutical composition according to claim 13 for use as a medicinal product for the prevention or treatment of neutrophilia in peripheral tissues. 20. Human antibody molecule according to any one of paragraphs. 1-12 or a pharmaceutical composition according to claim 13 for use in the prevention or treatment of airway neutrophilia or acute pneumonia. 21. The use of human antibody molecules according to any one of paragraphs. 1-12 or the pharmaceutical composition according to claim 13 when obtaining a medicinal product for the prevention or treatment of respiratory tract neutrophilia or acute pneumonia. 22. Use according to claim 21, wherein the airway neutrophilia, acute pneumonia, or both is chronic obstructive pulmonary disease, severe neutrophilic asthma, or both. 23. A method for blocking neutrophil chemotaxis, including exposing neutrophils to a human antibody according to any one of claims. 1-12. 24. The method according to claim 23, wherein chemotaxis is the migration of neutrophils into the lungs. 25. A method of blocking CXCR2 signaling in response to CXCL1 and/or CXCL5 in cells expressing CXCR2 signaling, comprising exposing the cells to a human antibody according to any one of claims. 1-12.