A method for specifically eliminating adipocytes that present perilipin-1 fragments

Abstract

The present invention relates to antigen-binding peptides that specifically bind to at least one perilipin-1 epitope as defined herein. The present invention further relates to antigen-binding peptides for use in the prevention or treatment of obesity. Furthermore, the present invention relates to bispecific antigen-binding peptides comprising a first binding site that specifically binds to a perilipin-1 epitope and a second binding site, and uses thereof. The present invention also relates to pharmaceutical compositions comprising the antigen-binding peptides or bispecific antigen-binding peptides for use in the prevention or treatment of obesity. The present invention further relates to isolated nucleic acids encoding the antigen-binding peptides or bispecific antigen-binding peptides. Furthermore, the present invention relates to recombinant cells comprising the nucleic acids, and methods for producing them as means for specifically targeting perilipin-1 fragment-presenting adipocytes.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to antigen-binding peptides that specifically bind to at least one perilipin-1 epitope as defined herein. The present invention further relates to antigen-binding peptides for use in the prevention or treatment of obesity. Furthermore, the present invention relates to bispecific antigen-binding peptides comprising a first binding site that specifically binds to a perilipin-1 epitope and a second binding site, and uses thereof. The present invention also relates to pharmaceutical compositions comprising the antigen-binding peptides or bispecific antigen-binding peptides for use in the prevention or treatment of obesity. The present invention further relates to isolated nucleic acids encoding the antigen-binding peptides or bispecific antigen-binding peptides. Furthermore, the present invention relates to recombinant cells comprising the nucleic acids, and to methods for producing means for specifically targeting perilipin-1 fragment-presenting adipocytes. [Background technology]

[0002] Background of the Invention Obesity is a global problem, and according to the World Health Organization, over 1.9 billion people are overweight. Obese individuals are commonly susceptible to a variety of medical conditions. While surgical intervention is currently a successful therapeutic approach, it often results in complications associated with subsequent gastrointestinal problems. Therefore, surgical intervention is considered acceptable only in severely obese patients with high comorbidities. Relatively promising pharmacological therapies have been forced to withdraw from the market due to severe side effect profiles. The multifaceted nature of the signaling mechanisms involved has led to unacceptable side effects. Therefore, there is a lack of therapeutic approaches that are successful over the long term and have negligible side effects.

[0003] The harmful long-term effects of excess body fat are widely known. However, there are also medical conditions, namely, lipodystrophy, that result in the loss of body fat. While metabolic complications vary with the degree of fat loss, common consequences of lipodystrophy include insulin resistance or diabetes mellitus, the absence of ketosis, an increased metabolic rate, hyperlipidemia, and hepatomegaly, including fatty liver. These outcomes, as demonstrated in various animal models, result from the absence of metabolically active adipocytes, regardless of the underlying cause of adipocyte loss. Lipodystrophy has attracted widespread attention as a common side effect of some antiretroviral drugs, which cause partial redistribution of fat.

[0004] Perilipin-1 is specifically expressed in adipocytes but not in other cells that store triacylglycerol, such as hepatocytes. [1] In these other cells, other members of the lipid droplet-associated protein family, such as adipophilin and TIP47, are present in place of perilipin-1. Because perilipin-1 shields lipid storage droplets in adipocytes from the activity of lipolytic enzymes, any mechanism that results in depletion of lipid storage droplet-associated perilipin-1 can lead to increased lipolysis and release of lipids stored in the droplets (and thereby worsen metabolic conditions). Therefore, a reduction in fat mass is expected at the expense of unfavorable metabolic conditions, which can be prevented by completely eliminating the adipocytes. Because other types of lipid-storing cells do not display perilipin-1, targeting perilipin-1 to activate immune effector mechanisms against adipocytes can enable their specific elimination. WO02 / 028410A1 discloses that perilipin may be a target for regulating body weight, muscle mass, and diabetes. Greenberg et al. (J. Biol. Chem., 1991, vol. 266, no. 17, pp. 11341-11346) describe perilipin-1 and disclose that it is present intracellularly on the surface of lipid droplets in adipocytes, such as lipid storage droplets.

[0005] Autoimmune responses against adipocytes result in the near-complete loss of adipose tissue, as observed in generalized acquired lipodystrophy (AGL) [2]. There is extensive clinical evidence for an autoimmune etiology in many cases of classical acquired lipodystrophy. Serum autoantibodies against adipose tissue have been found in patients with acquired generalized lipodystrophy [2]. In particular, autoantibodies against perilipin-1 have been found to be associated with acquired generalized lipodystrophy [3].

[0006] Therefore, the effector system that causes fat loss in lipodystrophy patients can be used to specifically target adipose tissue in obese patients.However, such a specific effector system, which can be, for example, an antibody or antigen-binding fragment or cytotoxic T cell, has not been obtained.In particular, despite great efforts, no adipocyte-specific epitope that can potentially be targeted as a specific therapeutic target has been identified so far [4]. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 02 / 028410 [Non-patent literature]

[0008] [Non-Patent Document 1] Greenberg AS, Egan JJ, Wek SA, Moos MC, Jr., Londos C, Kimmel AR. Isolation of cDNAs for perilipins A and B: sequence and expression of lipid droplet proteins associated of adipocytes. Proc Natl Acad Sci USA, 1993;90:12035-9. [Non-patent document 2] Greenberg et al. (J. Biol. Chem., 1991, vol. 266, no. 17, pp.11341-11346 [Non-patent document 3] Huebler A, Abendroth K, Keiner T, Storcker W, Kauf E, Hein G et al. Dysregulation of insulin-like growth factors in a case of generalized acquired lipoatrophic diabetes mellitus (Lawrence Syndrome) connected with autoantibodies against adipocyte membranes. Exp Clin Endocrinol Diabetes, 1998;106:79-84. [Non-patent document 4] Corvillo F, Aparicio V, Lopez-Lera A, Garrido S, Araujo-Vilar D, et al. Autoantibodies against perilipin 1 as a cause of acquired generalized lipodystrophy. Front Immunol, 2018; 9:2142. [Non-Patent Document 5] Misra A, Garg A. Clinical features and metabolic derangements in acquired generalized lipodystrophy: case reports and review of the literature. Medicine (Baltimore), 2003;82:129-46. Summary of the Invention [Problem to be solved by the invention]

[0009] Therefore, an object of the present invention is to provide a means for specifically targeting and / or eliminating adipocytes, such as an antigen-binding peptide or cytotoxic T cells, etc. Furthermore, an object of the present invention is to provide a method for producing the means for specifically targeting adipocytes. [Means for solving the problem]

[0010] Summary of the Invention The present invention relates to epitopes of perilipin-1, which are molecular target structures for specifically targeting and / or eliminating adipocytes. In particular, the epitopes disclosed herein are target structures of the intracellular protein perilipin-1, which are presented on the surface of adipocytes. Antigen-binding peptides targeting the epitopes disclosed herein can be used alone or in combination with any therapeutic agent to activate immune effector systems, such as extrinsic or endogenous effector systems, to target and / or eliminate adipocytes. The epitopes disclosed herein are also useful for generating perilipin-1-specific cytotoxic T cells that can target perilipin-1 fragment-presenting adipocytes.

[0011] The present inventors disclose perilipin-1 as an autoantigen targeted by autoantibodies derived from the serum of AGL patients. Furthermore, the present inventors disclose various epitopes of perilipin-1 having any of SEQ ID NOS: 1-6, 330-333, and provide antibodies and antigen-binding fragments targeting the epitopes, which are useful in the treatment of obesity. The present inventors surprisingly found that perilipin-1 and / or fragments thereof are present on the surface of adipocytes, rather than simply covering or surrounding intracellular lipid droplets, such as lipid storage droplets, within the adipocytes. Without wishing to be bound by any theory, the present inventors have found that perilipin-1 and / or fragments thereof become presented or are otherwise present on the surface of adipocytes, and thus can be specifically targeted.

[0012] The present invention further relates to the production of means for specifically targeting adipocytes, which are useful in the treatment or prevention of obesity-related diseases and / or medical conditions.

[0013] The present invention aims to use exogenous or endogenous effector systems of the immune system to specifically target and / or eliminate adipocytes. Such exogenous effector systems can be, for example, antibodies and / or cytotoxic T cells generated and / or amplified ex vivo or in vitro. The epitopes of perilipin-1 disclosed herein, which are SEQ ID NOS: 1-6, 330-333, enable specific targeting of adipocytes via perilipin-1 fragments presented on the adipocytes, particularly those present on the surface of the adipocytes. Targeting the adipocytes via perilipin-1 using antigen-binding peptides specific for any of SEQ ID NOS: 1-6, 330-333 marks the adipocytes for effector systems such as the complement system, resulting in activation of FcRIII (CD16), Fc-RII (CD32), Fc-RI (CD64), or other complement and / or Fc-receptors, leading to recruitment and activation of effector systems such as ADCC (antibody-dependent cell-mediated cytotoxicity) by NK (natural killer) cells or ADCP (antibody-dependent cellular phagocytosis) by macrophages, neutrophils, or any other effector system bearing the respective Fc-receptor. Recruitment of one of these effector systems results in targeting and / or elimination of the adipocytes. Alternatively, the adipocytes can be directly targeted by cytotoxic T cells specific for perilipin-1, particularly any of SEQ ID NOS: 1-6, 330-333, which then eliminate the adipocytes presenting at least one of these perilipin-1 epitopes. Thus, the means for specifically targeting and / or eliminating adipocytes according to the present invention may, in the case of ADCP, directly eliminate the adipocytes and thus serve as a means for direct elimination, or may indirectly eliminate the adipocytes by targeting them, resulting in activation of effector systems such as the complement system, ADCC, or cytotoxic T cells and thus serve as a means for indirect elimination.

[0014] The cell type specificity of the effector response is ensured by the specificity of the cellular antigen, i.e., perilipin-1, in particular the epitope having an amino acid sequence according to any of SEQ ID NOS: 1 to 6, 330 to 333. Thus, the present invention provides a means for specifically targeting and / or eliminating adipocytes without off-target effects in other cells and thus with negligible side effects.

[0015] Furthermore, targeting of adipocytes for immune effector systems using antigen-binding peptides allows for systemic application, for example, by injection, and / or allows for fine-tuned, targeted administration to eliminate certain fat depots by administration to the respective fat depot, such as by injection into the targeted fat depot.

[0016] The present invention utilizes adipocyte-specific display of perilipin-1 fragments, typically intracellular proteins, i.e., proteins previously only known to exist intracellularly, on the adipocyte surface. Perilipin-1 is essential for lipid storage in adipocytes. The present inventors disclose herein six peptide sequences that represent epitopes in perilipin-1, identified by analyzing the binding patterns of anti-perilipin-1 autoantibodies from lipodystrophy patients. The present inventors also disclose additional peptide sequences that are variants of these epitopes, which contain additional residues and can function equally well as epitopes. Several of the identified epitopes are highly conserved in mouse and human perilipin-1 sequences.

[0017] The target antigen epitopes of perilipin-1 disclosed herein are: a) immunizing an animal with said target structure to produce polyclonal or monoclonal antibodies and / or antigen-binding fragments against perilipin-1 peptides presented by adipocytes; b) screening a recombinant antibody library and / or a small molecule library to identify specific antibodies, antigen-binding fragments or other suitable affinity ligands for perilipin-1 peptides presented by adipocytes, and / or c) It allows the isolation and / or generation in vitro of cytotoxic T cells specific for perilipin-1 peptides presented by adipocytes.

[0018] The adipocytes can be marked in vivo using antibodies, antibody fragments, or other suitable affinity ligands produced by the method according to a) or b), thereby inducing a specific immune effector response.Furthermore, adipocytes can be indirectly eliminated in vivo using cytotoxic T cells produced by the method according to c).

[0019] The elements of the present invention are described below. While these elements are listed by specific embodiments, it should be understood that they can be combined in any manner and in any number to create additional embodiments. The various described examples and preferred embodiments should not be construed as limiting the invention to only the specifically described embodiments. This description should be understood to support and encompass embodiments that combine two or more of the specifically described embodiments, or combine one or more of the specifically described embodiments with any number of disclosed and / or preferred elements. Furthermore, unless the context indicates otherwise, any permutation and combination of any described elements in this application should be considered to be disclosed by the description of this application.

[0020] In a first aspect, the present invention relates to an antigen-binding peptide that specifically binds to at least one perilipin-1 epitope selected from the following amino acid sequences: LTLLDGDLPE (SEQ ID NO: 1), EKIASELK (SEQ ID NO: 2), VPIASTSDKV (SEQ ID NO: 3), PAPGHQQAQK (SEQ ID NO: 4), PSLLSRVGALTN (SEQ ID NO: 5), and PWLHSLAA (SEQ ID NO: 6). In one embodiment, the at least one perilipin-1 epitope is selected from the following amino acid sequences: NKGLTLLDGDLPEQE (SEQ ID NO: 330), QYPPEKIASELKDTI (SEQ ID NO: 331), AKPSLLSRVGALTNT (SEQ ID NO: 332), and EVRVPWLHSLAAAQE (SEQ ID NO: 333).

[0021] In one embodiment, the antigen-binding peptide binds to two or more of SEQ ID NOs: 1 to 6 and SEQ ID NOs: 330 to 333.

[0022] In one embodiment, said perilipin-1 epitope is a human perilipin-1 epitope.

[0023] In one embodiment, the antigen-binding peptide is not a perilipin-1-targeting autoantibody from a human patient with lipoatrophy, and preferably is not a perilipin-1-targeting anti-PLIN1 antibody against a peptide spanning Thr8 to Ala145 having manufacturer R&D catalog #AF6615.

[0024] In one embodiment, the antigen-binding peptide competes for binding with perilipin-1-targeting autoantibodies from a human patient with lipoatrophy.

[0025] In one embodiment, the antigen-binding peptide has a binding affinity of at least 10 -4 Affinity of M (K D ) binds to perilipin-1.

[0026] In one embodiment, the antigen-binding peptide is an antibody, preferably a monoclonal antibody.

[0027] In one embodiment, the antigen-binding peptide is a polyclonal antibody.

[0028] In one embodiment, the antigen-binding peptide, particularly the antibody, when incubated with 3T3-L1 cells differentiated into adipocytes in the presence of complement, causes complement-mediated cell death of the 3T3-L1 cells and / or the differentiated adipocytes.

[0029] In one embodiment, the complement-mediated cell death is determined, preferably numerically, by determining the ratio of live cells to dead cells using a suitable dye or mixture of dyes, such as a fluorescent dye or mixture of fluorescent dyes. Typically, such a dye or mixture of dyes distinguishes between live and dead cells. An example of such a dye mixture is the dye commercially available from ThermoFisher as "FluoroQuench™ Staining / Quenching Reagent." Preferably, the 3T3-L1 cells are differentiated into adipocytes using a suitable differentiation medium, such as a serum-free differentiation (SFD) medium supplemented with insulin and, if necessary, further supplemented with dexamethasone and / or isobutyl-1-methylxanthine.

[0030] In one embodiment, the complement is selected from mammalian complement, preferably human complement or rodent complement. In one embodiment, the rodent complement is rabbit complement or mouse complement. Complement is commercially available through various companies, such as BAG Diagnostics GmbH, BAG Health Care, or Bio-Rad. Alternatively, serum or plasma can serve as a source of complement.

[0031] In one embodiment, when the antigen-binding peptide, particularly the antibody, is incubated with adipocytes, the antigen-binding peptide, particularly the antibody, binds to the surface of the adipocytes. In one embodiment, the binding of the antigen-binding peptide, preferably the antibody, to the surface of the adipocytes is determined by incubating the adipocytes with the respective antigen-binding peptide, preferably the respective antibody, and then detecting the respective antigen-binding peptide, preferably the respective antibody, when bound to the adipocytes using a suitable detection technique, such as immunochemical detection alone or in combination with flow cytometry.

[0032] The "surface" of the adipocyte, as used herein, typically refers to the outer surface of such adipocyte, i.e., the extracellular surface exposed to the surroundings of the adipocyte.

[0033] In one embodiment, the antigen-binding peptide, particularly the antibody, when incubated with 3T3-L1 cells differentiated into adipocytes in the presence of complement, causes complement-mediated cell death of the 3T3-L1 cells and / or the differentiated adipocytes, and also binds to the surface of the adipocytes after incubation with the adipocytes.

[0034] In one embodiment, the antigen-binding peptide is i. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 18, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 20, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 22, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 23, or ii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 24, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 25, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 28, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 29, or iii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 30, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 31, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 32, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 34, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 35, or iv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 36, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 37, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 38, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 40, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 41, or v. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 42, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 43, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 44, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 45, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 46, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 47, or vi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 48, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 49, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 50, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 51, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 52, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 53, or vii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 54, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 55, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 56, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 57, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 58, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 59; or viii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 60, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 61, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 62, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 63, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 64, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 65; or ix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 66, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 67, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 68, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 69, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 70, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 71; or x. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 72, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 73, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 74, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 75, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 76, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 77, or xi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 78, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 79, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 80, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 81, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 82, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 83; or xii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 84, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 85, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 86, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 87, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 88, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 89, or xiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 90, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 91, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 92, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 93, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 94, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 95, or xiv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 96, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 97, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 98, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 99, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 100, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 101, or xv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 102, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 103, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 104, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 105, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 106, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 107, or xvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 108, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 109, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 110, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 111, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 112, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 113; or xvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 114, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 115, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 116, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 117, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 118, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 119; or xviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 120, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 121, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 122, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 123, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 124, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 125; or xix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 126, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 127, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 128, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 129, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 130, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 131; or xx. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 132, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 133, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 134, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 135, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 136, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 137, or xxi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 138, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 139, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 140, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 141, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 142, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 143; or xxii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 144, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 145, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 146, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 147, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 148, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 149; or xxiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 150, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 151, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 152, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 153, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 154, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 155; or xxiv. A heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 156, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 157, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 158, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 159, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 160, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 161, or xxv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 162, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 163, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 164, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 165, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 166, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 167; or xxvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 168, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 169, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 170, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 171, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 172, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 173; or xxvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 174, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 175, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 176, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 177, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 178, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 179; or xxviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 180, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 181, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 182, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 183, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 184, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 185; or xxix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 186, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 187, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 188, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 189, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 190, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 191; or xxx. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 192, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 193, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 194, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 195, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 196, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 197; or xxxi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 198, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 199, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 200, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 201, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 202, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 203; or xxxii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 204, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 205, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 206, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 207, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 208, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 209; or xxxiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 210, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 211, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 212, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 213, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 214, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 215; or xxxiv. A heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 216, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 217, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 218, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 219, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 220, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 221, or xxxv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 222, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 223, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 224, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 225, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 226, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 227; or xxxvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 228, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 229, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 230, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 231, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 232, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 233; or xxxvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 234, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 235, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 236, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 237, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 238, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 239; or xxxviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 240, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 241, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 242, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 243, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 244, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 245; or xxxix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 246, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 247, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 248, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 249, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 250, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 251; or xl. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 252, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 253, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 254, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 255, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 256, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 257; or xli. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 258, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 259, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 260, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 261, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 262, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 263; or xlii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 264, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 265, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 266, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 267, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 268, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 269; or xliii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 270, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 271, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 272, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 273, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 274, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 275; or xliv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 276, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 277, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 278, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 279, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 280, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 281; or xlv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 282, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 283, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 284, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 285, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 286, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 287, or xlvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 288, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 289, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 290, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 291, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 292, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 293; or xlvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 294, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 295, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 296, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 297, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 298, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 299; or xlviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 300, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 301, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 302, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 303, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 304, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 305; or xlix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 306, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 307, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 308, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 309, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 310, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 311; or l. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 312, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 313, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 314, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 315, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 316, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 317; or a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 318, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 319, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 320, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 321, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 322, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 323; or lii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 324, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 325, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 326, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 327, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 328, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 329; an antigen-binding peptide, preferably an antibody; Or, it is a peptide, preferably an antibody, whose heavy chain CDR1, CDR2, and CDR3 regions and whose light chain CDR1, CDR2, and CDR3 regions comprise amino acid sequences that are at least 90% identical, preferably at least 95% identical, more preferably at least 98% identical, and even more preferably 99% identical to any of the sequences set forth in i. to lii.

[0035] In one embodiment, the antigen-binding peptide is i. comprising the amino acid sequence of SEQ ID NO: 334 and / or comprising the amino acid sequence of SEQ ID NO: 335; or ii. comprising the amino acid sequence of SEQ ID NO: 336 and / or comprising the amino acid sequence of SEQ ID NO: 337; or iii. comprising the amino acid sequence of SEQ ID NO: 338 and / or comprising the amino acid sequence of SEQ ID NO: 339; or iv. comprising the amino acid sequence of SEQ ID NO: 340 and / or comprising the amino acid sequence of SEQ ID NO: 341; or v. comprising the amino acid sequence of SEQ ID NO: 342 and / or comprising the amino acid sequence of SEQ ID NO: 343; or vi. comprising the amino acid sequence of SEQ ID NO: 344 and / or comprising the amino acid sequence of SEQ ID NO: 345; or vii. comprising the amino acid sequence of SEQ ID NO: 346 and / or comprising the amino acid sequence of SEQ ID NO: 347; or viii. comprising the amino acid sequence of SEQ ID NO: 348 and / or comprising the amino acid sequence of SEQ ID NO: 349; or ix. comprising the amino acid sequence of SEQ ID NO: 350 and / or comprising the amino acid sequence of SEQ ID NO: 351; or x. comprising the amino acid sequence of SEQ ID NO: 352 and / or comprising the amino acid sequence of SEQ ID NO: 353; or xi. comprising the amino acid sequence of SEQ ID NO: 354 and / or comprising the amino acid sequence of SEQ ID NO: 355; or xii. comprising the amino acid sequence of SEQ ID NO: 356 and / or comprising the amino acid sequence of SEQ ID NO: 357; or xiii. comprising the amino acid sequence of SEQ ID NO: 358 and / or comprising the amino acid sequence of SEQ ID NO: 359; or xiv. comprising the amino acid sequence of SEQ ID NO: 360 and / or comprising the amino acid sequence of SEQ ID NO: 361; or xv. comprising the amino acid sequence of SEQ ID NO: 362 and / or comprising the amino acid sequence of SEQ ID NO: 363; or xvi. comprising the amino acid sequence of SEQ ID NO: 364 and / or comprising the amino acid sequence of SEQ ID NO: 365; or xvii. comprising the amino acid sequence of SEQ ID NO: 366 and / or comprising the amino acid sequence of SEQ ID NO: 367; or xviii. comprising the amino acid sequence of SEQ ID NO: 368 and / or comprising the amino acid sequence of SEQ ID NO: 369; or xix. comprising the amino acid sequence of SEQ ID NO: 370 and / or comprising the amino acid sequence of SEQ ID NO: 371; or xx. comprising the amino acid sequence of SEQ ID NO: 372 and / or comprising the amino acid sequence of SEQ ID NO: 373; or xxi. comprising the amino acid sequence of SEQ ID NO: 374 and / or comprising the amino acid sequence of SEQ ID NO: 375; or xxii. comprising the amino acid sequence of SEQ ID NO: 376 and / or comprising the amino acid sequence of SEQ ID NO: 377; or xxiii. comprising the amino acid sequence of SEQ ID NO: 378 and / or comprising the amino acid sequence of SEQ ID NO: 379; or xxiv. comprising the amino acid sequence of SEQ ID NO: 380 and / or comprising the amino acid sequence of SEQ ID NO: 381; or xxv. comprising the amino acid sequence of SEQ ID NO: 382 and / or comprising the amino acid sequence of SEQ ID NO: 383; or xxvi. comprising the amino acid sequence of SEQ ID NO: 384 and / or comprising the amino acid sequence of SEQ ID NO: 385; or xxvii. comprising the amino acid sequence of SEQ ID NO: 386 and / or comprising the amino acid sequence of SEQ ID NO: 387; or xxviii. comprising the amino acid sequence of SEQ ID NO: 388 and / or comprising the amino acid sequence of SEQ ID NO: 389; or xxix. comprising the amino acid sequence of SEQ ID NO: 390 and / or comprising the amino acid sequence of SEQ ID NO: 391; or xxx. comprising the amino acid sequence of SEQ ID NO: 392 and / or comprising the amino acid sequence of SEQ ID NO: 393; or xxxi. comprising the amino acid sequence of SEQ ID NO: 394 and / or comprising the amino acid sequence of SEQ ID NO: 395; or xxxii. comprising the amino acid sequence of SEQ ID NO: 396 and / or comprising the amino acid sequence of SEQ ID NO: 397; or xxxiii. comprising the amino acid sequence of SEQ ID NO: 398 and / or comprising the amino acid sequence of SEQ ID NO: 399; or xxxiv. comprising the amino acid sequence of SEQ ID NO: 400 and / or comprising the amino acid sequence of SEQ ID NO: 401; or xxxv. comprising the amino acid sequence of SEQ ID NO: 402 and / or comprising the amino acid sequence of SEQ ID NO: 403; or xxxvi. comprising the amino acid sequence of SEQ ID NO: 404 and / or comprising the amino acid sequence of SEQ ID NO: 405; or xxxvii. comprising the amino acid sequence of SEQ ID NO: 406 and / or comprising the amino acid sequence of SEQ ID NO: 407; or xxxviii. comprising the amino acid sequence of SEQ ID NO: 408 and / or comprising the amino acid sequence of SEQ ID NO: 409; or xxxix. comprising the amino acid sequence of SEQ ID NO: 410 and / or comprising the amino acid sequence of SEQ ID NO: 411; or xl. comprising the amino acid sequence of SEQ ID NO: 412 and / or comprising the amino acid sequence of SEQ ID NO: 413; or xli. comprising the amino acid sequence of SEQ ID NO: 414 and / or comprising the amino acid sequence of SEQ ID NO: 415; or xlii. comprising the amino acid sequence of SEQ ID NO: 416 and / or comprising the amino acid sequence of SEQ ID NO: 417; or xliii. comprising the amino acid sequence of SEQ ID NO: 418 and / or comprising the amino acid sequence of SEQ ID NO: 419; or xliv. comprising the amino acid sequence of SEQ ID NO: 420 and / or comprising the amino acid sequence of SEQ ID NO: 421; or xlv. comprising the amino acid sequence of SEQ ID NO: 422 and / or comprising the amino acid sequence of SEQ ID NO: 423; or xlvi. comprising the amino acid sequence of SEQ ID NO: 424 and / or comprising the amino acid sequence of SEQ ID NO: 425; or xlvii. comprising the amino acid sequence of SEQ ID NO: 426 and / or comprising the amino acid sequence of SEQ ID NO: 427; or xlviii. comprising the amino acid sequence of SEQ ID NO: 428 and / or comprising the amino acid sequence of SEQ ID NO: 429; or xlix. comprising the amino acid sequence of SEQ ID NO: 430 and / or comprising the amino acid sequence of SEQ ID NO: 431; or l. comprising the amino acid sequence of SEQ ID NO: 432 and / or comprising the amino acid sequence of SEQ ID NO: 433; or 1. comprising the amino acid sequence of SEQ ID NO: 434 and / or comprising the amino acid sequence of SEQ ID NO: 435; or lii. comprising the amino acid sequence of SEQ ID NO: 436 and / or comprising the amino acid sequence of SEQ ID NO: 437; an antigen-binding peptide, preferably an antibody; Alternatively, the antigen-binding peptide is a peptide, preferably an antibody, comprising an amino acid sequence having at least 90% identity, preferably at least 95% identity, more preferably at least 98% identity, and even more preferably 99% identity to any of the sequences set forth in i. to lii.

[0036] The respective sequences are also outlined in Table 5 and detailed in the sequence listing and in Figure 16 (HCDR1-3 and LCDR1-3 sequences) and Figure 17 (VH- and VL-sequences).

[0037] The term "comprising," when used herein in connection with an amino acid sequence of a certain SEQ ID NO, is typically intended to allow for the presence of additional residues flanking such sequence, which additional residues are not listed in the respective SEQ ID NO. However, the term "comprising" is also intended to include the possibility that such sequence does not contain any additional residues (not listed in the respective SEQ ID NO), in which case the term "comprising" is used in the sense of "consisting" and should be understood as "consisting of." As an example, a region "comprising" the amino acid sequence of SEQ ID NO: xyz includes the sequence listed in SEQ ID NO: xyz, but can also include additional flanking residues not listed in SEQ ID NO: xyz; however, such terminology also includes the possibility that a region "consists" only of the amino acid sequence of SEQ ID NO: xyz, in which case it is intended that any additional flanking residues are not encompassed by or present in said region.

[0038] In one embodiment, the antigen-binding peptide is an antibody further selected from the antibodies shown in Table 5 below.

[0039] In one embodiment, the antigen-binding peptide is a human antibody, a humanized antibody, or a chimeric human antibody.

[0040] In one embodiment, the antigen-binding peptide is an intact antibody, a substantially intact antibody, a Fab fragment, a F(ab')2 fragment, or a single chain Fv fragment.

[0041] In one embodiment, the antigen-binding peptide is capable of binding to a perilipin-1 fragment displayed on an adipocyte, and the binding of the antigen-binding peptide to the perilipin-1 fragment on the surface of the adipocyte activates the immune effector system to target and / or eliminate the adipocyte.

[0042] In one embodiment, the antigen-binding peptide comprises an Fc region, preferably an Fc region that has been optimized by protein engineering for higher binding affinity to effector cells compared to the binding affinity of a non-optimized Fc region.

[0043] In one embodiment, the antigen-binding peptide does not cross-react with any other perilipin proteins, such as perilipin-2, -3, -4, or -5.

[0044] In a further aspect, the antigen-binding peptide is an antigen-binding peptide for use in the prevention or treatment of obesity.

[0045] The antigen-binding peptide is as defined above and elsewhere herein.

[0046] In a further aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (i) a first binding site that specifically binds to a perilipin-1 epitope having any one of SEQ ID NOs: 1 to 6, 330 to 333; (ii) a second binding site; and The present invention relates to a bispecific antigen-binding peptide comprising:

[0047] In one embodiment, the second binding site binds to a perilipin-1 epitope having any of SEQ ID NOs: 1-6, 330-333, which is equal to or different from the epitope of the first binding site; to a white adipocyte-specific surface marker, e.g., the neutral amino acid transporter ASC-1; or to an immune cell-specific receptor molecule, such as a T cell-specific receptor molecule or a natural killer cell-specific receptor molecule.

[0048] Said epitope is as defined in any of the embodiments above and elsewhere herein.

[0049] In a further aspect, the bispecific antigen-binding peptide is a bispecific antigen-binding peptide for use in the prevention or treatment of obesity.

[0050] In a further aspect, the present invention also relates to a composition comprising an antigen-binding peptide according to the invention as defined herein and / or a bispecific antigen-binding peptide according to the invention as defined herein.

[0051] In one embodiment, the composition comprises two or more antigen-binding peptides according to the invention as defined herein and / or two or more bispecific antigen-binding peptides according to the invention as defined herein.

[0052] In one embodiment, when a composition comprises two or more antigen-binding peptides and / or two or more bispecific antigen-binding peptides, the two or more antigen-binding peptides are a set of peptides, each of which binds to a different perilipin-1 epitope, and the two or more bispecific antigen-binding peptides are a set of bispecific peptides, each of which binds to a different perilipin-1 epitope.

[0053] In one embodiment, the composition comprises at least two, or at least three, or at least four, or at least five, or at least six antigen-binding peptides according to the invention as defined herein, each of which binds to a different perilipin-1 epitope, and / or at least two, or at least three, or at least four, or at least five, or at least six bispecific antigen-binding peptides according to the invention as defined herein, each of which binds to a different perilipin-1 epitope.

[0054] In a further aspect, the present invention also relates to a composition comprising two or more antigen-binding peptides according to the invention as defined herein and / or two or more bispecific antigen-binding peptides according to the invention as defined herein for use as a pharmaceutical composition in the prevention or treatment of obesity.

[0055] In a further aspect, the present invention relates to a pharmaceutical composition comprising said antigen-binding peptide and / or said bispecific antigen-binding peptide for use in the prevention or treatment of obesity.

[0056] The antigen-binding peptide and the bispecific antigen-binding peptide are as defined in any of the embodiments above and elsewhere herein.

[0057] In a further aspect, the invention relates to an isolated nucleic acid encoding said antigen-binding peptide or encoding said bispecific antigen-binding peptide.

[0058] The antigen-binding peptide and the bispecific antigen-binding peptide are as defined in any of the embodiments above and elsewhere herein.

[0059] In a further aspect, the invention relates to a recombinant cell comprising said nucleic acid, which is preferably a recombinant autologous effector cell or a recombinant host cell.

[0060] Said nucleic acid is as defined in any of the embodiments above and elsewhere herein.

[0061] In a further aspect, the present invention provides a method for producing a means for specifically targeting perilipin-1 fragment-presenting adipocytes, comprising the steps of: a) inoculating a non-human animal with an antigen comprising at least one perilipin-1-specific epitope selected from SEQ ID NOS: 1 to 6, 330 to 333, and screening and producing antigen-binding peptides that specifically bind to the at least one perilipin-1-specific epitope from the inoculated animal, preferably by collecting cells that express the antigen-binding peptide from the spleen of the non-human animal, preparing hybridoma cells therefrom, expressing the antigen-binding peptide, and screening the antigen-binding peptides expressed by the hybridoma cells to identify antigen-binding peptides that specifically bind to any of SEQ ID NOS: 1 to 6, 330 to 333, and isolates thereof; or b) providing and screening a recombinant antigen-binding peptide library, such as a library of antibodies, scFv molecules, camelid antibodies, anticalins or DARPins, to identify antigen-binding peptides that specifically bind to any of SEQ ID NOS: 1-6, 330-333 and isolates thereof; or c) providing and screening a cell pool containing cytotoxic T cells, such as by tetramer staining or dextramers techniques, to identify cytotoxic T cells having specificity for any of SEQ ID NOS: 1-6, 330-333 and isolates thereof; or d) providing a patient blood sample and obtaining T cells therefrom, and genetically modifying said T cells by introducing a nucleic acid as described above and elsewhere herein, and obtaining chimeric antigen receptor-expressing T cells; or e) providing and screening a pool of T cell receptors to identify T cell receptors having specificity for any of SEQ ID NOS: 1-6, 330-333 and isolates thereof; The present invention relates to a method comprising:

[0062] In one embodiment, the means targets perilipin-1 and is selected from an antigen-binding peptide and a cytotoxic T cell.

[0063] The antigen-binding peptide and the epitope are as defined in any of the embodiments above and elsewhere herein.

[0064] In a further aspect, the present invention also relates to the use of an antigen-binding peptide or a bispecific antigen-binding peptide in the manufacture of a pharmaceutical composition for the treatment or prevention of obesity.

[0065] The antigen-binding peptide, the bispecific antigen-binding peptide and the pharmaceutical composition are as defined above and elsewhere herein.

[0066] In a further aspect, the present invention also relates to the use of an antigen-binding peptide, a bispecific antigen-binding peptide or a specific cytotoxic T cell in the manufacture of a preparation, preferably a pharmaceutical preparation, for the treatment or prevention of obesity.

[0067] The antigen-binding peptide, the bispecific antigen-binding peptide and the specific cytotoxic T cells are as defined above and elsewhere herein.

[0068] In a further aspect, the present invention also relates to a method for preventing or treating obesity, comprising the step of administering to a patient in need thereof an antigen-binding peptide that specifically binds to any of the perilipin-1 epitopes having SEQ ID NOs: 1-6, 330, 333, a bispecific antigen-binding peptide and / or cytotoxic T cells.

[0069] In one embodiment, the patient is a human.

[0070] The antigen-binding peptide, the bispecific antigen-binding peptide and the cytotoxic T cells are as defined above and elsewhere herein.

[0071] In a further aspect, the present invention relates to the use of at least one perilipin-1 epitope having any of SEQ ID NOs: 1-6, 330-333 for the identification of antigen-binding peptides, bispecific antigen-binding peptides and / or cytotoxic T cells that are specific for adipocytes and / or can be used in methods for the treatment or prevention of obesity.

[0072] The antigen-binding peptide, the bispecific antigen-binding peptide and the cytotoxic T cells are as defined above and elsewhere herein. DETAILED DESCRIPTION OF THE INVENTION

[0073] Detailed Description The term "antigen-binding peptide," as used herein, relates to a peptide that specifically binds to an antigen, preferably a peptide that binds to a perilipin-1 epitope selected from any of SEQ ID NOS: 1-6. In one embodiment, the perilipin-1 epitope is selected from any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the antigen-binding molecule is based on an immunoglobulin, such as a polyclonal or monoclonal antibody, or on a protein scaffold with antigen-binding capacity, such as anticalin proteins, affilins, affimers, affitins, alphabodies, nanobodies, or DARPins. In one embodiment, the antigen-binding peptide relates to an antibody or antibody fragment. In one embodiment, reference to an antibody or antibody fragment refers to an antigen-binding peptide. There are several classes of antibodies, particularly human antibodies, such as IgA, IgG, IgM, IgD, and IgE, and subclasses such as IgG1, IgG2, IgG3, and IgG4. Antibodies are proteins produced primarily by plasma cells of the immune system to neutralize pathogens. Antibodies are grouped into classes, also called isotypes, genetically determined by their constant regions. Human constant light chains are classified as kappa (Cκ) and lambda (Cλ) light chains. Human heavy chains are classified as mu, delta, gamma, alpha, or epsilon, defining antibody isotypes as IgM, IgD, IgG, IgA, and IgE, respectively. The IgG class is most commonly used for therapeutic purposes. "IgG," as used herein, refers to a polypeptide belonging to the class of antibodies substantially encoded by recognized immunoglobulin gamma genes. In humans, this class includes subclasses IgG1, IgG2, IgG3, and IgG4. In mice, this class includes subclasses IgG1, IgG2a, IgG2b, and IgG3. IgA has several subclasses, including, but not limited to, IgA1 and IgA2.Thus, "isotype," as used herein, refers to any of the immunoglobulin classes or subclasses defined by the chemical and antigenic characteristics of their constant regions. Known human immunoglobulin isotypes are IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgM, IgD, and IgE. IgG, a hybrid composition of natural human IgG isotypes, may also be useful in the present invention. Effector functions, such as ADCC, ADCP, CDC, and serum half-life, vary significantly among different classes of antibodies, including, for example, human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD, IgE, and IgM. In one embodiment, the antigen-binding peptide can comprise domains from any of the above-mentioned antibody classes, subclasses, and / or chains. In one embodiment, the amino acid sequence of the antigen-binding peptide can be modified by protein engineering, for example, to include constant regions from other immunoglobulin classes that mediate improved effector function properties. Such engineered hybrid IgG compositions can confer improved effector function properties, including improved ADCC, phagocytosis, CDC and serum half-life.

[0074] In the present invention, the antigen-binding peptide can relate to, for example, a human antibody, a human chimeric antibody, a humanized antibody, or an antibody fragment. The antibody recognizes an antigen via a Fab fragment variable region containing a paratope. The paratope specifically targets an epitope in the antigen. In one embodiment, the antigen-binding peptide of the present invention binds to any of the epitopes in perilipin-1 having SEQ ID NOS: 1-6, 330-333. In one embodiment, the binding of the antigen-binding peptide to a perilipin-1 fragment displayed on the cell surface of an adipocyte results in activation of the immune effector system to eliminate the adipocyte. Furthermore, the term "antigen-binding peptide," as used herein, relates to a peptide that specifically binds to a target antigen, such as one or more epitopes displayed by or present on the target antigen. The antigen to which the antigen-binding peptides of the invention bind is perilipin-1, or in the case of bispecific molecules, perilipin-1 and, optionally, another antigen, such as an antigen on the surface of effector cells, such as an antigen on the surface of cytotoxic T cells, an antigen presented on the surface of adipocytes other than a perilipin-1 fragment, or an antigen presented by any component of adipose tissue. In one embodiment, the bispecific molecule comprises a first binding site that binds to one of the perilipin-1 epitopes having any of SEQ ID NOS: 1-6, 330-333, and a second binding site for the same perilipin-1 epitope as the first binding site or for a perilipin-1 epitope having any of SEQ ID NOS: 1-6, 330-333 that is different from the epitope of the first binding site. In one embodiment, the bispecific molecule comprises a first binding site that binds to one of the perilipin-1 epitopes having any of SEQ ID NOS: 1-6, 330-333, and a second binding site for an antigen different from perilipin-1.

[0075] In one embodiment, the antigen-binding peptide is a TCR-derived antigen-binding peptide related to a T cell receptor specific for any of SEQ ID NOS: 1-6, 330-333, or a fragment of a T cell receptor specific for any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the T cell receptor may be a soluble T cell receptor or may be expressed on the surface of a genetically modified cell, preferably a T cell. In one embodiment, the TCR-derived antigen-binding peptide is used in adoptive T cell receptor transfer. In one embodiment, the antigen-binding peptide is related to a peptide expressed on the surface of a genetically modified cell, particularly a T cell or an NK cell, which has been genetically modified to express the antigen-binding peptide, resulting in a CAR-T cell or a CAR-NK cell, respectively.

[0076] In the present invention, the antigen-binding peptide may further refer to a fragment of an antibody, such as a substantially intact antibody, a Fab fragment, a F(ab')2 fragment, a diabody, a single-chain Fv fragment, a tetrabody, a triabody, a disulfide-stabilized Fv (dsFv), or a camelid-derived heavy chain VHH fragment. However, other forms of antigen-binding peptides are also contemplated by the present invention. For example, the antigen-binding peptide may be another (non-antibody) receptor protein derived from a small and robust non-immunoglobulin scaffold, such as one equipped with a binding function by using combinatorial protein design methods. Such a non-antibody antigen-binding peptide may be, for example, an affibody molecule, an affilin, an affimer, an affitin, an alphabody, an anticalin, a nanobody, or a DARPin. The antigen-binding peptide may also refer to other antibody mimetic molecules, such as dual-affinity retargeting antibodies (DARTs). In one embodiment, the antigen-binding peptide preferably relates to an antibody or antibody fragment, hi one embodiment, the antigen-binding peptide can relate to any antibody mimetic molecule that mimics the binding properties of an antibody to an antigen, but is structurally distinct from naturally occurring antibodies.

[0077] In one embodiment, antigen-binding peptides can be optimized to have improved properties, such as higher binding affinity and longer serum half-life, compared to non-optimized antigen-binding peptides, by protein engineering methods known to those skilled in the art, such as directed evolution, phage display, or ribosome display. In one embodiment, antigen-binding peptides can be modified to have the ability to induce a specific effector function, such as by including an Fc region to be targeted by effector cells, by protein engineering methods known to those skilled in the art. In one embodiment, the optimized antigen-binding peptide can be engineered to increase the response of effector systems to the Fc region by exchanging specific amino acids in the amino acid sequence of the Fc region. In one embodiment, the optimized antigen-binding peptide can be optimized to have an improved serum half-life, for example, by modifying the CH3 region of the IgG domain. In one embodiment, the antigen-binding peptide can be modified by protein engineering methods known to those skilled in the art so as to improve its ability to induce a particular effector function, such as a stronger effector response by the complement system or cytotoxic T cells, when recognizing an optimized antigen-binding peptide bound to an epitope presented on or present on the surface of adipocytes, compared to a non-optimized antigen-binding peptide bound to an epitope presented on or present on the surface of adipocytes, the epitope having any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the antigen-binding peptide is a bispecific antigen-binding peptide.

[0078] The terms "complementarity-determining region," "CDR," and "hypervariable region," as used herein, refer to one or more hypervariable or complementarity-determining regions (CDRs) found in the variable regions of the light or heavy chains of an antibody. The CDRs in each chain are held in close proximity by framework regions and, together with the CDRs from the other chain, contribute to the formation of the antigen-binding site. In one embodiment, an antigen-binding peptide comprises at least one, and preferably all, CDRs of an antibody that specifically binds to a perilipin-1 fragment, preferably to a perilipin-1 epitope having any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the antigen-binding peptide comprises a peptide scaffold and at least one, and preferably all, CDRs of an antibody that specifically binds to a perilipin-1 fragment, wherein the peptide scaffold can be an antibody fragment or any non-antibody peptide scaffold, such as an anticalin, affilin, affimer, affitin, alphabody, nanobody, or DARPin.

[0079] The term "bispecific antigen-binding peptide," as used herein, refers to an artificial protein comprising a first binding site that specifically binds to a perilipin-1 epitope having any of SEQ ID NOS: 1-6, 330-333, and a second binding site. Bispecific antigen-binding peptides can simultaneously bind to two target structures, such as two different types of antigens. In one embodiment, the bispecific antigen-binding peptide is based on immunoglobulins. In one embodiment, the bispecific antigen-binding peptide comprises binding fragments based on two different immunoglobulins, each with binding specificity to an antigen, and the two fragments are connected by a linker fragment, such as a linker peptide sequence, and / or an antibody scaffold. In one embodiment, the binding fragments of a bispecific antigen-binding peptide can be specific for different antigens, different epitopes on the same antigen, or the same epitope on the same antigen. In one embodiment, a bispecific antigen-binding peptide of the present invention binds to a first perilipin-1 epitope and a second, but different, non-overlapping perilipin-1 epitope. In one embodiment, a bispecific antigen-binding peptide of the present invention binds to a perilipin-1 epitope and a different antigen. In one embodiment, the different antigen is a surface marker of effector cells, such as T cells or natural killer cells. In one embodiment, the different antigen may be an antigen presented on the surface of adipocytes other than a perilipin-1 fragment, such as the neutral amino acid transporter ASC-1. In one embodiment, the different antigen may be an antigen presented by any component of adipose tissue other than perilipin-1. In one embodiment, the bispecific antigen-binding peptide is a fusion protein comprising a TCR-derived antigen-binding peptide. In one embodiment, the bispecific antigen-binding peptide comprises a TCR-derived antigen-binding peptide as a first binding site and a second binding site derived from a molecule selected from an antibody fragment, an anticalin, a DARPin, an aptamer, an affibody molecule, an affilin, an affimer, an affitin, an alphabody, and a nanobody.In one embodiment, the second binding site specifically binds to a receptor on an immune cell such as a T cell or natural killer cell, preferably a receptor that can activate the immune cell or stimulate an immune response of the immune cell, where the immune response is preferably a cytotoxic immune response. In one embodiment, the receptor is preferably a receptor molecule such as CD3, CD3δ / ε, CD3γ / ε, TCR, TCRα, TCRβ, CD2, CD5, CD28, OX40, 4-1BB, CD16, Ly49, NKp30 (CD337), NKp44 (CD336), NKp46 (NCR1), or CD3ζ (CD247). In one embodiment, the second binding site of the bispecific antigen-binding peptide specifically binds to a second perilipin-1 epitope having any of SEQ ID NOs: 1-6, 330-33, which second perilipin-1 epitope is different from and does not overlap with the perilipin-1 epitope of the first binding site.

[0080] The present invention also relates to compositions of one or several antigen-binding peptides and / or bispecific antigen-binding peptides according to the invention, as well as uses of such compositions. For example, such compositions may comprise one antigen-binding peptide according to the invention, e.g., an antibody and / or one bispecific antigen-binding peptide according to the invention. As another example, such compositions may comprise a plurality of different antigen-binding peptides according to the invention, e.g., antibodies and / or a plurality of different bispecific antigen-binding peptides according to the invention. Such compositions of a plurality of different antigen-binding peptides according to the invention, e.g., antibodies and / or different bispecific antigen-binding peptides, may also be referred to herein as a "pool" of different antigen-binding peptides, e.g., antibodies and / or different bispecific antigen-binding peptides. Typically, in such a "pool," each of the different antigen-binding peptides, e.g., each of the antibodies and / or each of the different bispecific antigen-binding peptides, binds to a different perilipin-1 epitope as defined herein; for example, such a pool can be a pool of different antibodies, each of these different antibodies binding to a different perilipin-1 epitope as defined herein. The perilipin-1 epitope as defined herein is preferably an epitope having an amino acid sequence selected from SEQ ID NOs: 1-6, 330-333.

[0081] The term "autoantibody," as used herein, relates to an antibody produced by a patient's immune system and directed against one or more of the patient's own proteins. Many autoimmune diseases are caused by such autoantibodies. In one embodiment, anti-perilipin-1 autoantibodies are involved in acquired systemic lipodystrophy. In one embodiment, said anti-perilipin-1 autoantibodies induce complement-mediated destruction of adipocytes in acquired systemic lipodystrophy.

[0082] The term "lipodystrophy," as used herein, relates to a medical condition in which the body is unable to produce and maintain healthy adipose tissue. Different types of lipodystrophy exist, such as congenital lipodystrophy syndrome and acquired lipodystrophy syndrome. The term "lipodystrophy" typically describes the (near) complete loss of adipose tissue from an area of ​​the body or the entire body. In one embodiment, lipodystrophy is used synonymously with lipodystrophy. In one embodiment, lipodystrophy relates to acquired generalized lipodystrophy (AGL). In one embodiment, the present invention discloses anti-perilipin-1 autoantibodies involved in lipodystrophy.

[0083] The term "perilipin-1," as used herein, is also known as lipid droplet-associated protein and relates to a protein associated with the surface of lipid droplets. Perilipin-1 belongs to the perilipin family, a family of proteins associated with the surface of lipid droplets. Perilipin-1 expression is elevated in obese humans. Perilipin-1 protects lipid droplets from lipases involved in lipolysis. In humans, three perilipin-1 isoforms exist: perilipin-A, perilipin-B, and perilipin-C. The human amino acid sequence of perilipin-1 is set forth in SEQ ID NO: 7. In one embodiment, the term "perilipin-1" relates to the full-length perilipin-1 protein and / or fragments thereof.

[0084] The term "epitope," as used herein, refers to the portion of an antigen that is recognized by the immune system, particularly antibodies, B cells, or T cells. In one embodiment, epitope is used synonymously with antigenic determinant. The portion of an antibody that binds to an epitope is a paratope. An epitope can be a conformational epitope or a linear epitope, depending on its structure and interaction with the paratope. In one embodiment, the antigen perilipin-1 comprises six epitopes, each having one of the sequences disclosed in SEQ ID NOS: 1-6, 330-333. The antibody or antigen-binding fragment of the present invention can bind to one or more of SEQ ID NOS: 1-6, 330-333. In one embodiment, the antibody or antigen-binding fragment of the present invention can also bind to a sequence having at least 85% identity to any of SEQ ID NOS: 1-6, 330-333.

[0085] The term "antigen," as used herein, relates to a molecule capable of inducing an immune response in an organism. An antigen can be targeted by an antibody. An antigenic determinant of an antigen is a distinct surface feature, i.e., an epitope. Most antigens are potentially bound by multiple antibodies, each typically specific for one of the antigen's epitopes. Autoantigens are normal proteins of an organism that are recognized by the immune system of said organism suffering from a specific autoimmune disease. In one embodiment, the antigen relates to perilipin-1 protein and / or fragments thereof. In one embodiment, the antigen, particularly the perilipin-1 fragment, is presented by MHC molecules on the surface of adipocytes. In one embodiment, the MHC molecule is an MHC-I or MHC-II molecule.

[0086] The term "targeting," as used herein, refers to the ability of a molecular structure, such as an antibody, an antigen-binding peptide, or a cytotoxic T-cell surface molecule, to bind to a particular structure, such as an antigen, particularly an epitope, through a specific interaction. In one embodiment, perilipin-1 fragments on the surface of adipocytes are targeted by antigen-binding peptides, resulting in the activation of effector systems, which in turn result in the elimination of the adipocytes, for example, via the caspase cascade. In one embodiment, perilipin-1 fragments on the surface of adipocytes are targeted by cytotoxic T cells that specifically bind to the perilipin-1, preferably resulting in cell death of the adipocytes. In one embodiment, the terms "targeting" and "recognition" are used interchangeably. In one embodiment, the antigen-binding peptides of the present invention target perilipin-1 protein and / or perilipin-1 fragments, preferably via an epitope having any of SEQ ID NOs: 1-6, 330-333. In one embodiment, an antigen-binding peptide of the present invention targets a perilipin-1 fragment presented by MHC on the cell surface, or such a perilipin-1 fragment may be present or presented on the cell surface by another mechanism. In one embodiment, an antigen-binding peptide targets at least one perilipin-1 epitope having any of SEQ ID NOS: 1-6, 330-333. In one embodiment, targeting a perilipin-1 fragment presented on an adipocyte using an antigen-binding peptide preferably results in elimination of said adipocyte by an effector / elimination mechanism. In one embodiment, when the antigen-binding peptide binds to two or more of SEQ ID NOS: 1-6 and 330-333, the antigen-binding peptide preferably binds to any of the following double or triple combinations a)-s): a) SEQ ID NOs: 1 and 2; b) SEQ ID NOs: 1 and 5; c) SEQ ID NOs: 1 and 6; d) SEQ ID NOs: 2 and 5; e) SEQ ID NOs: 2 and 6; f) SEQ ID NOs: 5 and 6; g) SEQ ID NOs: 1, 2 and 5; h) SEQ ID NOs: 1, 2 and 6; i) SEQ ID NOs: 2, 5 and 6; k) SEQ ID NOs: 330 and 331; l) SEQ ID NOs: 330 and 332; m) SEQ ID NOs: 330 and 333; n) SEQ ID NOs: 331 and 332; o) SEQ ID NOs: 331 and 333; p) SEQ ID NOs: 332 and 333; q) SEQ ID NOs: 330, 331, and 332; r) SEQ ID NOs: 330, 331, and 333; and s) SEQ ID NOs: 331, 332, and 333.

[0087] Alternatively, the present invention also envisions that binding to more than one perilipin-1 epitope can be achieved by providing a composition (or "pool") of several different antigen-binding peptides in accordance with the invention as defined herein, each of which binds to a single perilipin-1 epitope that is distinct from the perilipin-1 epitopes bound by the other antigen-binding peptides in such composition (or "pool").

[0088] Thus, in one embodiment, such a composition may, for example, preferably comprise two or three antigen-binding peptides that bind to any of the double or triple combinations a) to s) described above.

[0089] The term "elimination," as used herein, refers to targeting and inducing cell destruction of target cells, for example, by the complement cascade, by recruitment of NK cells, or by attack from cytotoxic T cells, preferably further including degradation and / or removal of the resulting cellular debris. In one embodiment, the antigen-binding peptides of the invention induce the elimination of adipocytes by an effector mechanism. In one embodiment, targeted adipocytes displaying perilipin-1 fragments on their surface are eliminated by induction of apoptosis by cytotoxic T cells or by lysis of said cells by the complement cascade. In one embodiment, the term "elimination" can refer to cell damage, cell destruction, cell disintegration, and / or cell phagocytosis. In one embodiment, cell elimination refers to cell damage sufficient to terminate the cell's ability to survive and / or maintain its cellular function. In many embodiments, elimination preferably relates to the complete removal of adipocytes. In many embodiments, "elimination" refers to the destruction and complete degradation of adipocytes, preferably by phagocytosis, preferably without leaving extracellular debris. In one embodiment, perilipin-1 displayed on adipocytes is targeted using an antigen-binding peptide to eliminate the adipocytes. In one embodiment, adipocyte elimination can be triggered by one effector mechanism, by several effector mechanisms simultaneously, or by a cascade of effector mechanisms.

[0090] The term "effector mechanism," as used herein, relates to a mechanism involved in and / or resulting in the elimination of target cells. Effector mechanism, as used herein, can refer to various mechanisms of action involved in the destruction of target cells, such as activating death pathways in target cells, blocking necessary growth factor / growth factor receptor interactions, inducing antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC) responses, and / or antibody-dependent cellular phagocytosis (ADCP). In one embodiment, the effector mechanism is an immune effector mechanism, which is a mechanism of the immune system for eradicating cells, such as infected cells. In one embodiment, the term "effector mechanism" is used interchangeably with "effector system." In one embodiment, the term effector mechanism can refer to an individual effector mechanism involved in the elimination of adipocytes, or a group of several effector mechanisms involved in the elimination of adipocytes, either simultaneously or as a cascade of mechanisms.

[0091] In one embodiment, interaction of complement with the antigen-binding peptide elicits a complement-dependent cytotoxicity (CDC) response. In one embodiment, binding of the antigen-binding peptide to perilipin-1 fragments displayed on the surface of adipocytes mediates apoptosis of the adipocytes. In one embodiment, the antigen-binding peptide preferably comprises an Fc (fragment crystallisable)-derived region that mediates effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and / or antibody-dependent cellular phagocytosis (ADCP) through interaction with Fc receptors expressed by effector cells and / or components of the complement system, such as complement component 1q (C1q). In one embodiment, the antigen-binding peptide comprises an Fc region that contributes to the long half-life of the antigen-binding peptide mediated by binding to the salvage receptor FcRn. In one embodiment, glycosylation of the antigen-binding peptide can alter the strength of the effector function mediated by the antigen-binding peptide. In one embodiment, effector cells such as NK cells, monocytes, macrophages, and granulocytes recognize the antigen-binding peptides of the present invention bound to target cells, preferably adipocytes, and the recognition is carried out, for example, via the effector cell receptor FcγR and the Fc domain of the antigen-binding peptide, triggering the effector cells to release cytoplasmic perforin, granulysin, and granzymes, thereby inducing apoptosis and / or lysis of the target cells. In one embodiment, the effector mechanism leading to the destruction of adipocytes can also include a cytotoxic drug, toxin, or radionuclide delivered to the adipocytes by the antigen-binding peptides of the present invention, such as by conjugating a cytotoxic drug, toxin, or radionuclide to the antigen-binding peptide, or by using liposomes labeled with the antigen-binding peptide to deliver the cytotoxic drug, toxin, or radionuclide.

[0092] In one embodiment, the effector mechanism can involve a chimeric antigen receptor (CAR), a receptor protein engineered to confer the ability of T cells to target specific proteins. In particular, T cells are collected from a patient and genetically modified to contain a chimeric antigen receptor comprising an antigen-binding peptide of the present invention, for example, by transforming the T cells with a plasmid / virus containing the genetic information for a fusion protein comprising an antigen-binding peptide, such as a TCR-derived antigen-binding peptide, and a transmembrane domain and a cytoplasmic effector domain. The genetically modified T cells are then administered to the patient as CAR-T cells to attack adipocytes. In one embodiment, the CAR-T cells can be derived from T cells in the patient's own blood (autologous cells) or from T cells of another healthy individual (allogeneic cells). In one embodiment, the CAR comprises an antigen-binding peptide specific for any of SEQ ID NOS: 1-6, 330-333 as an ectodomain, as well as a transmembrane domain and an endodomain. In one embodiment, the antigen-binding peptide specific for any of SEQ ID NOS: 1-6, 330-333 can be a T cell receptor or a domain of a T cell receptor. In one embodiment, the effector mechanism can involve T cells genetically modified to be specific for any of SEQ ID NOS: 1-6, 330-333 and / or T cells naturally specific for any of SEQ ID NOS: 1-6, 330-333 selected from a pool of in vitro expanded T cells obtained from a donor, which can be autologous or allogeneic.

[0093] The term "effector cell," as used herein, refers to a cell, preferably an immune cell, involved in an effector system and / or mechanism that results in the destruction and / or elimination of target cells. In one embodiment, the effector cell can be a monocyte, macrophage, neutrophil, NK cell, NKT cell, or cytotoxic T cell. In one embodiment, the effector cell recognizes and / or binds to an antigen-binding peptide of the present invention bound to a perilipin-1 fragment presented on an adipocyte, recognizes and / or binds to a perilipin-1 fragment presented on an adipocyte, or recognizes and / or binds to a complex thereof. In one embodiment, the recognition and / or binding induces an effector mechanism that results in the elimination / destruction of the adipocyte. In one embodiment, the effector cell, such as a T cell, recognizes an adipocyte target cell and induces cell death of the target cell. In one embodiment, effector cells such as NK cells, monocytes, macrophages or granulocytes recognize the antigen-binding peptide of the present invention bound to adipocytes, and this recognition triggers the induction of apoptosis or lysis of the adipocytes and / or phagocytosis of the adipocytes.

[0094] The term "adipocyte," as used herein, refers to a fat cell specialized for fat storage. Adipose tissue is primarily composed of adipocytes. Different types of adipose tissue exist, namely, white fat, brown fat, beige fat, and marrow fat, and therefore different types of adipocytes also exist. White fat cells are monovacuolar cells containing a single large lipid droplet surrounded by a layer of cytoplasm. Brown fat cells are plurivacuolar and typically polygonal in shape. Beige fat contains white and brown fat cells, making it metabolically more desirable than white fat. Adipocytes in marrow adipose tissue also exist and are also thought to play a role in obesity. In one embodiment, the terms "perilipin-1-presenting adipocytes" and "perilipin-1 fragment-presenting adipocytes" are used interchangeably. In one embodiment, adipocytes preferably present a fragment of perilipin-1 on their surface. In one embodiment, the fragment of perilipin-1 displayed on the surface of an adipocyte preferably comprises any one of SEQ ID NOs: 1-6, 330-333.

[0095] The term "direct elimination mechanism," as used herein, relates to the process and / or ability of the immune effector system to directly destroy target cells and remove their constituents. In one embodiment, activation of macrophages through Fc receptor-mediated binding of an antigen-binding peptide to perilipin-1 or a fragment thereof results in destruction of target cells and removal of their constituents. In one embodiment, the direct elimination mechanism involves neutrophils (neutrophilic polymorphonuclear granulocytes) through Fc receptor-mediated binding of an antibody or antigen-binding fragment to perilipin-1 or a fragment thereof. In one embodiment, the direct elimination mechanism preferably involves macrophages and / or neutrophils binding to an antigen-binding peptide attached to a perilipin-1 fragment on the surface of an adipocyte, said binding resulting in phagocytosis of said adipocyte, said binding preferably being mediated by an Fc receptor.

[0096] The term "indirect elimination mechanism," as used herein, relates to the process and / or ability of the immune effector system to target a target cell and involve another system to destroy the target cell and / or remove the cell remnants. In one embodiment, an indirect elimination mechanism involves binding of an antigen-binding peptide to a target structure, such as perilipin-1 or a fragment thereof, on the surface of an adipocyte. In one embodiment, the binding triggers the destruction of the adipocyte by activation of the complement cascade. In one embodiment, the mechanism is complement-dependent cytotoxicity (CDC). In one embodiment, an indirect elimination mechanism involves antibody-dependent cell-mediated cytotoxicity (ADCC), which involves lysis of target cells by effector cells of the immune system, such as natural killer cells. In one embodiment, indirect elimination involves antibody-dependent cellular phagocytosis (ADCP), which involves engulfment of adipocytes by effector cells of the immune system, such as macrophages. In one embodiment, the indirect elimination mechanism involves cytotoxic T cell-mediated cytotoxicity, which involves lysis of adipocytes by effector cells that directly bind to perilipin-1 fragments presented via MHC on the adipocyte surface. This destruction of adipocytes then triggers the activation of other systems, such as phagocytosis, which result in the elimination of the destroyed adipocyte debris. In one embodiment, the indirect elimination mechanism involves the engulfment of target cell remnants by macrophages or neutrophils (neutrophilic polymorphonuclear granulocytes). In one embodiment, a means for specifically targeting and / or eliminating adipocytes can have both direct and indirect elimination capabilities. In one embodiment, macrophages and neutrophils are capable of both direct elimination, such as by Fc receptor-dependent binding to and engulfment of target adipocytes, and indirect elimination, such as by engulfment of destroyed adipocyte debris targeted by a different elimination mechanism, such as complement-mediated opsonization. In one embodiment, the direct and indirect elimination mechanisms can occur simultaneously and / or overlap in scope. In one embodiment, the term "effector mechanism" can refer to both indirect and direct elimination mechanisms. In one embodiment, the terms "effector mechanism" and "elimination mechanism" can be used interchangeably.

[0097] The term "effector system" or "immune effector system," as used herein, relates to any system involved in the immune system that can potentially contribute to an immune response. The immune effector system can relate, for example, to the complement system, T cells, such as cytotoxic T cells or regulatory T cells, natural killer cells, B cells, plasma cells, or macrophages. In one embodiment, the effector system relates to the complement system, macrophages, neutrophils, NK cells, NKT cells, and / or cytotoxic T cells. In one embodiment, the effector system is triggered by binding of an antigen-binding peptide to perilipin-1 presented on adipocytes, which triggers the elimination of the adipocytes.

[0098] The term "endogenous effector system," as used herein, refers to an endogenous immune effector system. An endogenous effector system can be, for example, the complement system, which can induce targeted cell destruction after activation, such as by opsonization. An endogenous effector system can also refer to endogenous cytotoxic T cells activated by specific antigens. An endogenous effector system can also refer to macrophages, or neutrophils, such as neutrophilic polymorphonuclear granulocytes, or NK cells.

[0099] The term "exogenous effector system," as used herein, refers to an immune effector system that is exogenous. An exogenous effector system can be, for example, cytotoxic T cells that are expanded ex vivo and then administered to a patient. Such ex vivo expanded cytotoxic T cells can be obtained from the patient or a donor, or from a commercially available source. In one embodiment, the exogenous effector system can be chimeric antigen receptor T cells (CAR-T cells), which are genetically engineered T cells that express a T cell receptor specific for any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the exogenous effector system can also be an exogenous T cell receptor (TCR), which can be a soluble TCR or TCR expressed by an effector cell, such as a T cell, preferably a T cell transformed to express a TCR specific for any of SEQ ID NOS: 1-6, 330-333.

[0100] The term "complement system," as used herein, refers to the part of the innate immune system that enhances the ability of antibodies and phagocytes to remove damaged cells from an organism. More than 30 proteins are involved in the complement system. The complement system can induce different immune functions, such as phagocytosis by opsonizing antigens, inflammation by attracting macrophages and neutrophils, and membrane attack. The classical complement pathway typically requires antigen-antibody complexes for activation. The complement system can involve a membrane attack complex, which forms a transmembrane channel and causes osmotic lysis of target cells. Additionally, macrophages aid in the removal of complement-coated target cells.

[0101] The term "cytotoxic T cells," as used herein, refers to T lymphocytes capable of killing cancer cells, infected cells, or otherwise damaged cells. T cells typically express a T cell receptor (TCR) that specifically recognizes an antigen. Such antigens are typically presented on the surface of cells by MHC-I molecules. If the T cell receptor of a cytotoxic T cell is specific for an antigen, it binds to the complex of the antigen and the MHC-I molecule and destroys the cell. The interaction of T cells with the complex of the antigen and the MHC-I molecule of a target cell typically also involves a co-receptor, i.e., CD8. In one embodiment, upon binding to such a complex on the surface of a target cell, T cells release any of the cytotoxins perforin, granzyme, and granulysin. In one embodiment, perforin activates granzyme, which enters the cytoplasm of the target cell, and its serine protease function triggers the caspase cascade, resulting in apoptosis of the target cell. In one embodiment, apoptosis of said target cells can also be induced by cell surface interactions between cytotoxic T cells and target cells, such as through Fas-Fas ligand interactions.

[0102] The term "means for specifically targeting perilipin-1 fragment-presenting adipocytes," as used herein, refers to any means capable of specifically targeting perilipin-1 fragments presented on the surface of adipocytes and directly and / or indirectly inducing the elimination of said adipocytes. In one embodiment, the means for specifically targeting perilipin-1 fragment-presenting adipocytes relates to an antigen-binding peptide capable of specifically recognizing a perilipin-1 fragment, preferably via an epitope having any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the means for specifically targeting perilipin-1 fragment-presenting adipocytes comprises and / or induces an effector mechanism that induces the elimination of said adipocytes. In one embodiment, the means for specifically targeting perilipin-1 fragment-presenting adipocytes relates to cytotoxic T cells having a T cell receptor specific for perilipin-1, preferably specific for an epitope of perilipin-1 having any of SEQ ID NOS: 1-6, 330-333. In one embodiment, the means for specifically targeting perilipin-1 fragment-presenting adipocytes induces an indirect and / or direct elimination mechanism. In one embodiment, the means for specifically targeting perilipin-1 fragment-presenting adipocytes is preferably a means for specifically eliminating perilipin-1 fragment-presenting adipocytes. In one embodiment, the means is capable of eliminating perilipin-1 fragment-presenting adipocytes and / or capable of inducing an effector mechanism that eliminates perilipin-1-presenting adipocytes.

[0103] The terms "specific" or "specifically binds," as used herein, mean that an antibody or immunoreceptor fragment, according to the present invention, is capable of specifically interacting with and / or binding to a specific antigen or set of specific antigens, but does not essentially bind to other antigens. Such binding can be illustrated by the specificity of the lock and key principle.

[0104] The term "nucleic acid," as used herein, relates to a nucleotide sequence such as a ribonucleic acid or deoxyribonucleic acid. In one embodiment, the nucleic acid encodes an antigen-binding peptide according to other embodiments of the invention. In one embodiment, the nucleic acid encodes a sequence that is at least 50%, preferably at least 85%, identical to a nucleic acid sequence encoding an antigen-binding peptide according to other embodiments of the invention. In one embodiment, the nucleic acid that is at least 50%, preferably at least 85%, identical to a nucleic acid sequence encoding an antigen-binding peptide will, due to redundancy in the genetic code, result in the same peptide product as a 100% identical sequence.

[0105] The term "non-human animal", as used herein, relates to animals that are not human, preferably mammals.

[0106] The term "vaccination", as used herein, relates to the administration of an antigen, preferably having at least one epitope having a sequence according to any of SEQ ID NOs: 1-6, 330-333, for immunization and production of antibodies or antigen-binding fragments specific for said antigen.

[0107] The term "screening" as used herein relates to the analysis of certain candidate molecules or cells for their binding specificity to a certain antigen, preferably perilipin-1. The goal of such screening is to specifically and with high affinity, preferably at least 10 -9 M, or for TCR-derived antigen-binding fragments, preferably at least 10 -4 The goal is to identify molecules, such as antigen-binding peptides, or cells, such as cytotoxic T cells, that bind to a particular antigen, preferably perilipin-1, with an affinity of M. In one embodiment, the best candidates identified by screening are then amplified, preferably in vitro, for example by cell culture expansion.

[0108] The term "host cell," as used herein, relates to a cell that can be used to produce the antigen-binding peptides of the present invention. In one embodiment, the host cell allows for the production of antibodies or antigen-binding fragments, and includes, for example, yeast cells or Chinese hamster ovary (CHO) cells. In one embodiment, the host cell is transformed to contain a nucleic acid encoding the antigen-binding peptide of the present invention, and the transformed host cell is referred to as a recombinant cell. In one embodiment, the host cell is an autologous or allogeneic effector cell, such as a T cell or an NK cell, which can be transformed into a CAR-T cell or a CAR-NK cell, respectively. In one embodiment, such a cell is a genetically modified cell that contains a nucleic acid encoding an antigen-binding peptide specific for perilipin-1.

[0109] The term "obesity," as used herein, relates to a medical condition in which excess body fat accumulates in the body to an extent that may have a negative effect on the health of the body. In one embodiment, adipocytes in obese patients are targeted and / or eliminated using means for specifically eliminating perilipin-1 fragment-presenting adipocytes, such as perilipin-1 fragment-specific antigen-binding peptides or perilipin-1-specific cytotoxic T cells.

[0110] The term "binding assay," as used herein, relates to an assay or analytical procedure used to study the binding of an antigen-binding peptide to an antigen. In one embodiment, a binding assay can refer to flow cytometry, tetramer staining, Western blot, fluorescence assay, surface plasmon resonance, or other binding assays known to those skilled in the art. In one embodiment, a binding assay (e.g., tetramer staining) can also relate to an assay used to study the binding of cytotoxic T cells, preferably the T cell receptor of said cytotoxic T cells, to an antigen. In one embodiment, a binding assay can relate to surface plasmon resonance measurement.

[0111] The term “K D" as used herein relates to the dissociation constant, which is the reciprocal of the association constant. In one embodiment, the antigen-binding peptides of the invention have a dissociation constant of at least 10 -4 Affinity of M (K D In one embodiment, the antigen-binding peptide is a peptide selected from an antibody, an antibody-derived fragment, and a non-antibody antigen-binding peptide, such as an anticalin or a DARPin, and has at least 10 -7 M, preferably at least 10 -9 M, more preferably at least 10 -11 Affinity of M (K D In one embodiment, the antigen-binding peptide is a TCR-derived antigen-binding peptide and binds to an antigen at a distance of at least 10 -4 Affinity of M (K D ) binds to antigen and / or MHC-antigen complexes. In one embodiment, the term "at least 10 -4 Affinity of M (K D )" is 10 -5 M, 10 -6 M or 10 -7 M et al., 10 -4 M or smaller K D It is intended to refer to a value.

[0112] The term "monoclonal antibody" or "mAB," as used herein, refers to an antibody obtained from a population of antibodies that are substantially identical based on their amino acid sequence. Monoclonal antibodies are typically highly specific. Furthermore, in contrast to polyclonal antibody preparations, which typically contain different antibodies directed against different epitopes of an antigen, mAbs are typically directed against a single epitope on an antigen. In addition to their specificity, mAbs are advantageous in that they can be synthesized by cell culture, such as by hybridoma cell cultures or recombinant host cells free of other immunoglobulin contaminants. In one embodiment, a mAb refers to a chimeric, humanized, or human antibody or antibody fragment. In one embodiment, the monoclonal antibody may be an optimized monoclonal antibody with an engineered Fc region with improved binding properties to be targeted by effector cells. In one embodiment, the optimized monoclonal antibody may be engineered by specific amino acid exchanges in the amino acid sequence of the Fc region. In one embodiment, the optimized monoclonal antibody may also be optimized to have an improved serum half-life, for example, by modification of the CH3 region of the IgG.

[0113] The term "human antibody," as used herein, relates to an antibody or fragment thereof that comprises only human sequences. In one embodiment, the human antibody is produced by display techniques, such as by phage display or ribosome display.

[0114] The term "humanized antibody," as used herein, refers to an immunoglobulin chain or fragment thereof, such as a Fab fragment, a Fab' fragment, a F(ab')2 fragment, an Fv fragment, or other antigen-binding fragment of an antibody, that contains sequences derived from a non-human immunoglobulin. In one embodiment, a humanized antibody is a human antibody in which certain CDR residues have been replaced by CDR residues of non-human origin having the desired specificity and affinity characteristics.

[0115] The term "chimeric human antibody," as used herein, refers to an artificial antibody designed by combining sequences from different species, such as mouse and human, through genetic or protein engineering. In one embodiment, a human chimeric antibody is a hybrid protein of an antigen-binding domain derived from a mouse antibody and the constant domains of a human antibody.

[0116] The term "substantially intact antibody," as used herein, relates to an antibody that has not been fragmented, truncated, or otherwise shortened and that retains its ability to specifically bind to an antigen.

[0117] The term "cross-react," as used herein, refers to off-target binding to a structure that is not the target structure. In one embodiment, "cross-reacting" refers to binding of an antibody, antigen-binding fragment, or cytotoxic T cell to a protein that is not perilipin-1, such as perilipin-2, -3, -4, or -5.

[0118] The term "(%) sequence identity" as used herein relates to the percentage of pairwise identical residues for a homology alignment of the sequence in question with the sequence of a polypeptide / nucleic acid of the present invention.

[0119] The term "patient" as used herein can relate to a human or an animal. The blood sample for obtaining T cells and / or antibodies can be an autologous sample or an allogeneic sample.

[0120] The term "recombinant antigen-binding peptide library," as used herein, refers to a pool of antigen-binding peptides produced by protein engineering methodologies known to those skilled in the art. For example, such a pool of antigen-binding peptides can be produced and screened by phage display, yeast display, mRNA display, or ribosome display. Display libraries display single-chain variable domain antigen-binding fragments, such as scFvs, or Fab fragments and contain encoding DNA or RNA. The genetic diversity of these libraries is typically generated by cloning repertoires of immunoglobulin heavy and light chain variable gene segments from naive or immunized individuals, or by randomizing antigen-binding fragments using synthetic DNA, or by a combination of these approaches, optionally combined with error-prone PCR. In one embodiment, antigen-binding peptides can be optimized to enhance the effector function and / or half-life of the antigen-binding peptides.

[0121] The term "optimized," as used herein, refers to the enhanced properties of a peptide modified by protein engineering compared to the unmodified form of the peptide. In one embodiment, an antigen-binding peptide is optimized with respect to the strength and / or type of effector function it elicits using an effector system by engineering the antigen-binding peptide to include an Fc region that is recognized by an Fc receptor of, for example, macrophages, monocytes, neutrophils, NK cells, or another effector cell. In one embodiment, the Fc region is optimized by genetic engineering to contain amino acid exchanges and / or altered glycosylation compared to the unmodified Fc region. In one embodiment, the serum half-life of an antigen-binding peptide is increased by engineering the peptide to include a half-life-increasing CH3 region of an IgG domain or by modifying the antigen-binding peptide with other half-life-increasing molecules.

[0122] The invention will now be further described by reference to the following drawings: [Brief explanation of the drawings]

[0123] [Figure 1] Labeling of lipid droplet membranes with IgG autoantibodies from a patient with Lawrence syndrome / AGL. Visualization, with the exception of fluorescent labeling, was achieved using 10 nm gold particles in electron microscopy (EM) and 1 nm gold particles in light microscopy (LM). A) Light microscopy of a univacuolar fat cell from human white adipose tissue. Silver-enhanced gold particles appear only in the membrane of the fat vacuole, which cannot be distinguished from the cell membrane at this magnification. The loose connective tissue between the fat cells is unstained. Bar, 25 μm. B) Fluorescence microscopy of a multivacuolar 3T3-L1 cell. Labeling was visualized with the cyanine dye Cy3. Bar, 50 μm. C, D) Transmission electron microscopy (TEM) of a 3T3-L1 cell. The surface of lipid storage droplets is densely labeled with gold particles. Only isolated gold particles are present in the cell membrane (white arrowheads), while gold particles are absent from the nuclear membrane. Bar, 1 μm. E) No labeling is present in 3T3-L1 cells at the preadipocyte stage. Bar, 1 μm. F) Freeze-fracture replica of a 3T3-L1 cell. Gold labeling appears in the phospholipid layer (P-face, white arrowhead). Labeling is absent on the convex inner droplet surface (E-face, black arrowhead) and in the core of the LSD. Bar, 0.3 μm. G) Freeze-fracture replica of the outer cytoplasmic fracture surface of the plasma membrane of a 3T3-L1 cell, characterized by elevated caveolae. Gold particles are not visible. Bar, 0.2 μm. H) TEM of mouse adipose tissue. Gold particles are deposited on the surface of the LSD (white arrowhead). The dashed line indicates the adipocyte plasma membrane. Nearby are tissue eosinophilic granulocytes. C, caveolae; core, lipid core of LSD; EF, outer cytoplasmic cleavage plane; EOSG, eosinophil granule; LSD, lipid storage droplet; N, nucleus; PM, plasma membrane. Bar, 0.5 µm. Images shown are representative of at least three independent experiments. [Figure 2]Human anti-adipose autoantibodies react with multiple perilipin-1 variants. A) Two-dimensional separation of rat adipocyte membrane proteins. In the first dimension, proteins were separated on a linear pH gradient IPG strip (pH 4-7). In the second dimension, proteins were separated using a 10% (w / v) homogeneous SDS-PAGE gel. In the overlay, spots detected by Coomassie staining are shown in green. Spots detected by anti-adipose autoantibodies in parallel blots are color-coded red. Immunoreactive spots are identified by their yellow color (marked with arrows). Spot numbers refer to the identifications listed in Table B. B) Overview of peptide mass fingerprint (PMF) analysis of protein spots. Three of the five analyzed spots were clearly identified as perilipin A (spots 1, 2, and 4), and two spots were identified as perilipin B (spots 4 and 5), respectively, resulting from differential splicing of the perilipin-1 transcript. Accession numbers are sequence identification numbers in GenBank. The theoretical molecular masses, shown in the fourth column, were calculated from the amino acid sequence. The Mascot interface (MASCOT 2.1.03) was used to display PMF spectra for identification and to plot Mascot scores (P = 0.05). C) Comparison of a commercial anti-perilipin-1 antibody (Affinity BioReagents, Golden, CO, USA) with human anti-lipid IgG. Ten micrograms of membrane protein were subjected to SDS-PAGE, and immunoblotting was performed after cutting the PVDF membrane between the two lanes of membrane protein (↓). Incubation of the patient's autoantibodies with the anti-perilipin-1 antibody demonstrates the same pattern of positive bands. [Figure 3]Autoantibodies do not interfere with lipid storage homeostasis in differentiated human preadipocytes. Human preadipocytes were induced to undergo adipose conversion and accumulate triacylglycerol over 14 days. These cells were loaded with fluorescently labeled fatty acids (NBD-FAs) in the presence of 10 nM insulin for 3 h. After washing to remove excess NBD-FAs, cells were incubated for an additional 3 h (A, C, E) or 15 h (B, D, F) in the presence of 10 nM insulin in the absence of antibodies (white bars) or in the presence of anti-adipose IgG (black bars) or control IgG (gray bars). Cells were harvested, total lipids were extracted, and aliquots were analyzed by thin-layer chromatography, with subsequent quantification of labeled intracellular lipids by fluorescence scanning. Results shown are the mean ± SEM of four experiments. A, B) Anti-adipose IgG does not interfere with the accumulation of NBD-FAs in the total cellular lipid pool, presented as relative fluorescence units (RFU). C, D) Autoantibodies have no effect on the distribution of incorporated NBD-FAs between pools of stored triacylglycerols. E, F) Autoantibodies have no effect on the distribution of incorporated NBD-FAs between pools of lipid synthesis and degradation intermediates, diacylglycerol (DAG) and monoacylglycerol (MAG). [Figure 4]Anti-adipose autoantibodies accumulate within cells with atypical morphology (dead cells) in the presence of serum. Exposure of 3T3-L1 adipocytes to either control IgG or anti-adipose IgG in serum-free culture medium resulted in barely detectable labeling of the cells when the cells were thoroughly washed before fixation and immunofluorescence detection (A, F). However, anti-adipose IgG (G–J), but not control IgG (B–E), resulted in immunoreactivity of a substantial proportion of cells in the presence of goat serum (not heat-inactivated). This effect was strongly dose-dependent. Closer examination revealed that under these conditions, anti-adipose IgG did not visibly accumulate on the surface of lipid droplets (N), but did accumulate within cells with atypical morphology (arrowheads; lipid droplets, staining not associated with condensed chromatin), an accumulation that is only rarely found in normal cultures of 3T3-L1 adipocytes. Note the typical staining pattern of lipid droplets around the nuclei of cells fixed and stained according to the standard protocol (L, M). (K) There was no immunoreactivity in fixed cells with control IgG. (A-J, N) Live cells incubated with primary antibody; (K-M) Fixed cells incubated with primary antibody. (M, N) Overlay of the indicated areas of the fluorescent images (J and L, respectively) and the corresponding phase-contrast images; the red channel was set to 2x intensity for better visibility in the overlaid images. Scale bar: 250 μm (A-L) or 100 μm (M-N). [Figure 5]Anti-adipose autoantibodies induce complement-dependent cell death in 3T3-L1 cells. 3T3-L1 adipocytes were generated in a serum-free differentiation system and allowed to accumulate lipids for up to 8 days after differentiation induction. Cells were preincubated with anti-adipose IgG or control IgG for 40 minutes, as indicated. Next, medium was added without (control) or with rabbit complement (final dilution of either 1:64 or 1:32), as indicated in the figure. Anti-adipose IgG was additionally tested at twice the normally used concentration in the presence of 1:32 complement (anti-adipose IgG 2x). A) Cells were fixed and stained with FluoroQuench AO / EB reagent, which results in green staining of live cells and orange staining of dying and dead cells (in flat adipocytes, lipid droplets appear unstained, whereas in round adipocytes, lipid droplets are only visible). Scale bar for all images: 100 μm. B) Quantification of three independent experiments indicates a significant number of dead cells in the presence of anti-adipose IgG plus complement, but not under control conditions. Significant differences relative to all respective controls were identified by Student-Newman-Keuls all-pairwise multiple comparison ANOVA (P<0.005); **, P<0.01; ***, P<0.001. C) Percentage of viable cells relative to the corresponding untreated control after 1 hour (Student's t-test; #, P<0.05). [Figure 6]Scheme of perilipin-1-dependent antibody-induced complement activation. Mechanism of action of the induced complement response against adipocytes mediated by anti-perilipin-1 antibodies. Perilipin-1 peptides are degraded by the proteasome, and the resulting perilipin-1 fragments are presented by MHC on the cell surface of adipocytes. Antibodies targeting epitopes having any of the peptide sequences, i.e., SEQ ID NOS: 1-6, 330-333, disclosed herein, specifically target adipocytes, enabling their specific elimination by complement activation (indirect elimination mechanism). Alternatively, cytotoxic T cells specific for at least one perilipin-1 epitope can eliminate adipocytes presenting perilipin-1 through the indirect elimination mechanism. In addition, other effector systems that can be activated by antibodies binding to perilipin-1 fragment-loaded MHC can use the direct elimination mechanism. These effector systems activated by Fc receptor-mediated binding to the adipocyte-binding antibody can be macrophages or neutrophils (neutrophilic polymorphonuclear granulocytes). [Figure 7] Anti-adipose autoantibodies bind to lipid-associated proteins. Lipid-associated proteins (1) and cytosolic proteins (2) from isolated rat fat pad adipocytes were separated by SDS-PAGE and stained with Coomassie blue (A) or transferred to a PVDF membrane (B). Anti-adipose IgG reacts with two large and several small bands in preparations containing lipid-associated proteins. (C) A control membrane cut along the molecular weight standard lane (↓) was incubated with either control serum followed by Protein A / G-AP (control IgG) or Protein A / G-AP alone (Protein A / G-AP), indicating that recognition of the bands in (B) was specific to the anti-adipose antibody. S, molecular weight standard. Data are representative of five experiments with rat fat pads; similar results were obtained with in vitro differentiated human adipocytes and mouse 3T3-L1 cells. [Figure 8]A small fraction of cells is susceptible to autoantibody-induced cell death. Cells were treated in parallel with the experiment summarized in Figure 5, but incubation was prolonged for 24 h. In addition, data obtained with a higher complement concentration (final dilution 1:16) are shown (A). Quantification of three independent experiments indicates significant cell loss in the presence of anti-adipose IgG plus complement, but not under control conditions. When comparing the number of dead cells (B) and viable cells (C), there is a significant difference due to the detachment of dead cells. (B) Significant differences relative to all respective controls were identified by Student-Newman-Keuls ANOVA with all pairwise multiple comparisons (p = 0.016); *, P < 0.05. (C) Percentage of viable cells relative to the corresponding untreated control after 24 h (Student t-test; #, P < 0.05; ##, P < 0.005). [Figure 9]Peptide array for perilipin-1 epitopes. Perilipin-1 epitopes were identified using peptide arrays (PEPperPRINT, PEPperMAP) with perilipin-1 autoantibodies, with SEQ ID NOs: 1-6, 330-333. Human perilipin-1 sequences were extended with neutral linker sequences at the N- and C-termini to prevent truncated peptides. The extended antigen sequences were translated into peptide sequences with 15 amino acids each, with a 14-amino acid overlap. The peptide arrays thus generated were screened with perilipin-1 autoantibodies as described in previous examples. Antibody specificity was verified by screening all possible 15-mer peptide sequences of perilipin-1-related proteins perilipin-2, -3, -4, and -5, which are involved in different functions of lipid storage and / or other cells. Therefore, cross-reactivity with other members of the perilipin family was excluded. A) Analysis of peptide arrays using human anti-perilipin-1 autoantibodies to identify perilipin-1 epitopes. Only perilipin-1 peptides were bound specifically and with high affinity. B) Quantification of antibody binding. Six peptides were bound with high affinity. Binding of the peptides depicted on the right (PEKEPPKP from perilipin-3 and PILVERPEP from perilipin-5) was negligible. Several identified peptides (epitopes) are highly conserved in the mouse and human sequences. [Figure 10-1]Representative flow cytometry fluorescence histograms (dark gray profiles) of anti-perilipin antibodies binding to primary adipocytes. Antibodies 17-20 (A), 21, 22, 24, and 25 (B) were analyzed. Controls represent cells treated with an isotype control and a secondary antibody (light gray profiles). A commercially available antibody against perilipin-1 (ab192716) from Abcam was used as a control (C). The overlapping histograms observed using the commercially available antibody (C) indicate that the commercially available anti-perilipin-1 antibody does not specifically bind to adipocytes. In contrast, the antibodies of the present invention show binding to adipocytes (shift of the histograms to the right) compared to the isotype control (A, B). [Figure 10-2] Representative flow cytometry fluorescence histograms (dark gray profiles) of anti-perilipin antibodies binding to primary adipocytes. Antibodies 17-20 (A), 21, 22, 24, and 25 (B) were analyzed. Controls represent cells treated with an isotype control and a secondary antibody (light gray profiles). A commercially available antibody against perilipin-1 (ab192716) from Abcam was used as a control (C). The overlapping histograms observed using the commercially available antibody (C) indicate that the commercially available anti-perilipin-1 antibody does not specifically bind to adipocytes. In contrast, the antibodies of the present invention show binding to adipocytes (shift of the histograms to the right) compared to the isotype control (A, B). [Figure 11-1] Representative flow cytometry fluorescence histograms of anti-perilipin antibodies binding to primary adipocytes (dark gray profile). Antibodies 26 to 32 of the present invention were analyzed. Controls represent cells treated with an isotype control and secondary antibody (light gray profile). Antibodies of the present invention bind more effectively to adipocytes than the isotope control. [Figure 11-2] Representative flow cytometry fluorescence histograms of anti-perilipin antibodies binding to primary adipocytes (dark gray profile). Antibodies 26 to 32 of the present invention were analyzed. Controls represent cells treated with an isotype control and secondary antibody (light gray profile). Antibodies of the present invention bind more effectively to adipocytes than the isotope control. [Figure 12-1] Complement-mediated cell death mediated by anti-perilipin antibodies. Complement-mediated cell death of 3T3-L1 cells differentiated into adipocytes after the addition of a commercially available antibody against perilipin-1 (ab192716) from Abcam, compared with the control (A) and after the addition of a single anti-perilipin antibody (B). The live / dead cell ratio was calculated from the fluorescence intensity of the fluorescent dye that stains only live cells divided by the fluorescence intensity of the dye that stains dead cells. The detected signal is compared with that of the isotope control. The signal obtained for the isotope control is indicated by a dashed line. The commercially available antibody tested (PLN1-Ab Abcam) was not significantly different from the signal of the isotype control, indicating no significant complement-mediated cell death (not significant: ns) (A). In contrast, the antibody of the present invention shows significant (*: p<0.05; **: p<0.01) mediation of complement-mediated cell death (Figure 12 panel (B)). Figure 12 panel (B) also shows the signal obtained in this experiment relative to the isotope control, again indicated by the dashed line. The indicated antibodies and their target epitopes as well as their respective SEQ ID NOs are listed in Table 5. [Figure 12-2]Complement-mediated cell death mediated by anti-perilipin antibodies. Complement-mediated cell death of 3T3-L1 cells differentiated into adipocytes after the addition of a commercially available antibody against perilipin-1 (ab192716) from Abcam, compared with the control (A) and after the addition of a single anti-perilipin antibody (B). The live / dead cell ratio was calculated from the fluorescence intensity of the fluorescent dye that stains only live cells divided by the fluorescence intensity of the dye that stains dead cells. The detected signal is compared with that of the isotope control. The signal obtained for the isotope control is indicated by a dashed line. The commercially available antibody tested (PLN1-Ab Abcam) was not significantly different from the signal of the isotype control, indicating no significant complement-mediated cell death (not significant: ns) (A). In contrast, the antibody of the present invention shows significant (*: p<0.05; **: p<0.01) mediation of complement-mediated cell death (Figure 12 panel (B)). Figure 12 panel (B) also shows the signal obtained in this experiment relative to the isotope control, again indicated by the dashed line. The indicated antibodies and their target epitopes as well as their respective SEQ ID NOs are listed in Table 5. [Figure 13-1] Complement-mediated cell death mediated by anti-perilipin antibodies. Fluorescence microscopy images of complement-mediated cell death of 3T3-L1 cells differentiated into adipocytes after the addition of anti-perilipin antibodies such as antibody nos. 20 and 25 (B), compared to a negative control (no antibody added) and an isotype control antibody (A). In the presence of anti-perilipin antibodies of the present invention, particularly antibody nos. 20 or 25, the intensity of live cell staining is decreased and the number of dead cells is increased. Thus, the antibodies of the present invention exhibit effective complement-mediated cell death. [Figure 13-2] Complement-mediated cell death mediated by anti-perilipin antibodies. Fluorescence microscopy images of complement-mediated cell death of 3T3-L1 cells differentiated into adipocytes after the addition of anti-perilipin antibodies such as antibody nos. 20 and 25 (B), compared to a negative control (no antibody added) and an isotype control antibody (A). In the presence of anti-perilipin antibodies of the present invention, particularly antibody nos. 20 or 25, the intensity of live cell staining is decreased and the number of dead cells is increased. Thus, the antibodies of the present invention exhibit effective complement-mediated cell death. [Figure 14]Complement-mediated cell death compared with commercially available antibodies. The mean (plus SEM) difference in the number of dead cells determined with and without complement is shown; the numbers above the bars indicate the P value of a two-tailed Student's t-test. With the exception of aF-Ab (a patient-derived polyclonal antibody), all tested commercially available antibodies showed non-significant results. The commercially available anti-perilipin antibodies tested were PA1-1052 (ThermoFisher), AF6615 (R&D Systems), CST Antibody #3470 (D418), and PA5-18693 (ThermoFisher). Therefore, commercially available antibodies do not mediate complement-mediated cell death. [Figure 15-1] The patient-derived anti-fat antibody (aF-AB) and antibody #5 bind to the adipocyte surface, whereas the commercial antibody PA1-1052 does not. (A) The contrast between the total cell overview on the left and the lipid vacuole representation on the right indicates that adipocytes are considered. (B) 10x magnification of a 50 μm counting section. Left: microvilli; right: AB-conjugated immunogold particles (white dots). (A+B) Cells incubated with aF-AB. At least 7-8 individual cells were included for counting. (C) Quantitative evaluation. ANOVA (P=0.00002) with multiple comparisons (Holm-Sidak, P values ​​in the figure). Increased gold particle binding compared to the control is observed for the patient-derived anti-fat antibody (aF-AB) and antibodies of the present invention, such as antibody #5. In contrast, no significant increase in gold particle binding compared to the control is observed for the commercial antibody PA1-1052. Thus, the antibodies of the present invention exhibit superior binding efficacy to adipocytes compared to commercially available anti-perilipin antibodies. [Figure 15-2]The patient-derived anti-fat antibody (aF-AB) and antibody #5 bind to the adipocyte surface, whereas the commercial antibody PA1-1052 does not. (A) The contrast between the total cell overview on the left and the lipid vacuole representation on the right indicates that adipocytes are considered. (B) 10x magnification of a 50 μm counting section. Left: microvilli; right: AB-conjugated immunogold particles (white dots). (A+B) Cells incubated with aF-AB. At least 7-8 individual cells were included for counting. (C) Quantitative evaluation. ANOVA (P=0.00002) with multiple comparisons (Holm-Sidak, P values ​​in the figure). Increased gold particle binding compared to the control is observed for the patient-derived anti-fat antibody (aF-AB) and antibodies of the present invention, such as antibody #5. In contrast, no significant increase in gold particle binding compared to the control is observed for the commercial antibody PA1-1052. Thus, the antibodies of the present invention exhibit superior binding efficacy to adipocytes compared to commercially available anti-perilipin antibodies. [Figure 16-1] Exemplary antigen-binding peptides of the present invention, particularly antibodies, and their respective CDR sequences. The CDR amino acid sequences of antibodies having antibody numbers 5 to 11, 13 to 14, and 16 to 58 are depicted. The CDR sequences of the heavy chain (HCDR) and light chain (LCDR) are shown. CDR1, 2, and 3 (HCDR) of the heavy chain are depicted from left to right and separated by "_". CDR1, 2, and 3 (LCDR) of the light chain are depicted from left to right and separated by "_". [Figure 16-2] Exemplary antigen-binding peptides of the present invention, particularly antibodies, and their respective CDR sequences. The CDR amino acid sequences of antibodies having antibody numbers 5 to 11, 13 to 14, and 16 to 58 are depicted. The CDR sequences of the heavy chain (HCDR) and light chain (LCDR) are shown. CDR1, 2, and 3 (HCDR) of the heavy chain are depicted from left to right and separated by "_". CDR1, 2, and 3 (LCDR) of the light chain are depicted from left to right and separated by "_". [Figure 17-1]Exemplary antigen-binding peptides of the invention, particularly antibodies, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies with antibody numbers 5-11, 13-14, and 16-58 are depicted. [Figure 17-2] Exemplary antigen-binding peptides of the invention, particularly antibodies, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies with antibody numbers 5-11, 13-14, and 16-58 are depicted. [Figure 17-3] Exemplary antigen-binding peptides of the invention, particularly antibodies, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies with antibody numbers 5-11, 13-14, and 16-58 are depicted. [Figure 17-4] Exemplary antigen-binding peptides of the invention, particularly antibodies, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies with antibody numbers 5-11, 13-14, and 16-58 are depicted. [Figure 17-5] Exemplary antigen-binding peptides of the invention, particularly antibodies, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies with antibody numbers 5-11, 13-14, and 16-58 are depicted. [Example]

[0124] Example 1 Isolation of autoantibodies targeting perilipin-1. Written informed consent was obtained from all tissue sample donors in accordance with a protocol approved by the Ethics Committee of Jena University Hospital. Stromal vascular cell populations (termed human preadipocytes) from adipose tissue samples were isolated and cultured in Medium 199 containing 10% (v / v) fetal bovine serum until confluence was achieved. [5] 3T3-L1 cells were obtained from ATCC and maintained in DMEM containing 10% (v / v) fetal bovine serum until confluence was achieved. All experiments involving differentiation or stimulation of 3T3-L1 cells were performed in serum-free medium (phenol red-free and with 7.5 mM HEPES, pH 7.2) consisting of DMEM and Ham's F12 medium in a 3:1 ratio, supplemented with gentamicin (40 μg / ml), fetuin (300 μg / ml), transferrin (2 μg / ml), pantothenate (17 μM), biotin (1 μM), and insulin (1 μM) [6]. For adipose conversion of 3T3-L1 cells, this medium was further supplemented with dexamethasone (100 nM) and 3-isobutyl-1-methylxanthine (500 μM). Unless otherwise indicated, all experiments involving anti-adipose IgG and live cells (including complement activation assays) were performed in the absence of serum. Immunoglobulin G (IgG) from 5 ml patient serum (anti-fat IgG) or control serum (control IgG) was purified on thiophilic agarose (AFFI-T™; Kem-En-Tec, Taastrup, Denmark) according to the manufacturer's instructions.

[0125] Example 2 Preparation and analysis of adipocyte lipid-associated proteins. Male Wistar rats were used. A suspension of adipocytes in PBS was thoroughly shaken with half a volume of chloroform. Centrifugation (2500 × g) at 25°C for 5 minutes separated the lipids dissolved in chloroform from the cytoplasmic proteins in the supernatant and from the lipid-associated proteins in the fat cake at the interface between the organic and aqueous phases. Extraction of the fat cake with chloroform was repeated twice. Proteins in the supernatant were precipitated with acetone at -20°C for 1 hour. The lipid-associated proteins were suspended in PBS by vortexing. Residual chloroform in the final pellet was evaporated in vacuo. 100 μg of lipid-associated or cytoplasmic proteins from adipocytes were solubilized in 100 μl sample buffer containing 8 M urea. 10 μl of the sample was separated on a minigel at 4°C. Gels were fixed and stained with Coomassie blue, or proteins were transferred to PVDF membranes (Immobilon-P, 0.45 μm, Millipore, Bedford) using a semi-dry blotting unit for 2 hours at 1 mA / cm² at 8°C. After staining with Ponceau S for 2 minutes, the membranes were washed with 0.1 N NaOH, rinsed with water, blocked overnight in 0.2% I-block (TROPIX, Bedford, MA) in Tris-buffered saline-Tween® 20 (TBS-T), and incubated with primary antibodies for 1 hour: anti-fat IgG and control IgG at a dilution of 1:1000, and polyclonal rabbit anti-perilipin-1 at a dilution of 1:1250. Protein G-alkaline phosphatase (AP) was diluted 1:5000. Anti-rabbit IgG-AP was diluted 1:1250. Blots were developed using the nitroblue tetrazolium method. Proteomic analysis of fat cake proteins was performed.

[0126] Example 3 Assessment of storage lipid metabolism. Confluent monolayers of human preadipocytes were induced for adipose conversion in serum-free medium containing 1 μM insulin, 1 μM cortisol, 500 μM 3-isobutyl-1-methylxanthine, and 1 μM rosiglitazone for 8 days, followed by triacylglycerol accumulation in the presence of insulin alone until day 14. Cellular pools of fatty acid-containing lipids, primarily triacylglycerols, were labeled with the fluorescent fatty acid analog 12-(N-(7-nitrobenzo-2-oxa-1,3-diazol-4-yl)amino)dodecanoic acid (NBD-FA) and analyzed by thin-layer chromatography and fluorescence imaging. Briefly, labeling was performed in serum-free medium containing 1% (w / v) fatty acid-free bovine serum albumin and 100 μM NBD-FA. After washing to remove excess NBD-FA, cells were incubated in the absence or presence of antibodies for various periods of time. The incubation was terminated by removal of the medium and immediate extraction of the lipids with 2-propanol.

[0127] Example 4 Labeling of lipid droplet membranes by IgG autoantibodies from lipoatrophic patients. Serum containing autoantibodies against adipose tissue from an AGL patient was used to identify the target autoantigen. A purified IgG fraction of the autoimmune serum (anti-adipose IgG) exclusively labeled adipose tissue, but not other tissues examined. Labeling was localized to adipocytes in immunohistochemically analyzed tissue sections (Figure 1A). In addition to human unilocular adipocytes, anti-adipose IgG stained adipocytes from all other mammalian species examined, but not chicken or carp adipocytes (data not shown). Immunofluorescent staining of in vitro differentiated 3T3-L1 mouse adipocytes containing multiple lipid storage droplets (LSDs) (Figure 1B) suggested specific reactivity of anti-adipose IgG with the surface of LSDs. Such specific labeling of the LSD surface was confirmed by transmission electron microscopy in immunogold-labeled ultrathin sections of differentiated 3T3-L1 cells (Figure 1C). The adipocyte plasma membrane was only sporadically labeled (Fig. 1D), and undifferentiated cells showed no labeling at all (Fig. 1E), suggesting that autoantigen expression is almost exclusively restricted to the LSD of adipocytes. These findings were confirmed in ultrathin sections from mouse adipose tissue (Fig. 1H).

[0128] Example 5 Anti-fat autoantibodies react with lipid-associated proteins identified as multiple perilipin-1 variants. We prepared delipidated and cytosolic fractions of lipid-associated proteins using isolated rat epididymal fat pad adipocytes. In Western blot experiments using anti-adipose IgG, a characteristic pattern of two large and several small bands was found almost exclusively in the lipid-associated protein fraction (Figure 7B). Neither IgG from healthy controls nor protein A / G-AP alone reacted with these bands (Figure 7C), indicating that the recognition of this band pattern is specific to anti-adipose autoantibodies. Thus, rat adipocyte fractions confirmed the results of microscopic analysis. After confirming that anti-adipose IgG specifically labeled putative adipocyte-derived autoantigen(s), we began to identify the autoantigens. After separation of the lipid-associated protein fraction by two-dimensional electrophoresis, several groups of immunoreactive spots were identified by anti-adipose IgG (Figure 2A). These spots were isolated and subjected to trypsin digestion, and the peptides were subsequently analyzed by MALDI-TOF-MS. A search of the NCBI protein database using mass spectrometry data identified all immunoreactive spots as variants of perilipin-1 (Figure 2B). Direct comparison of anti-adipose IgG and a commercially available anti-perilipin-1 antibody revealed identical staining patterns in Western analysis, confirming perilipin-1 as an adipocyte-specific autoantigen in AGL (Figure 2C).

[0129] Example 6 In the presence of serum, anti-fat autoantibodies accumulate in cells with atypical morphology. To gain information about the mechanism by which anti-fat IgG induces adipocyte loss, we assessed whether anti-fat IgG interferes with stored lipid metabolism in intact cells. Therefore, cells were labeled with fluorescent fatty acids (NBD-FAs), which are substrates for both lipogenic and lipolytic enzymes. Neither control IgG nor anti-fat IgG altered the total amount of labeled lipids during 3 or 15 h of incubation (Figure 3A, B). More specific examination of stored triacylglycerols (Figure 3C, D) and the lipolytic intermediates diacylglycerol and monoacylglycerol (Figure 3E, F) also revealed no effect of anti-fat IgG in in vitro differentiated human adipocytes. Therefore, in accordance with the almost exclusive localization of perilipin-1 on the surface of LSDs (Figure 1), it is unlikely that anti-fat IgG interferes with the normal function of perilipin-1 to induce adipocyte loss. However, when live 3T3-L1 adipocytes were incubated with the antibody in the absence or presence of increasing amounts of goat serum, a dose-dependent increase in the number of labeled cells could be detected in the presence of anti-adipose IgG (Fig. 4F–J), but not with control IgG (Fig. 4A–E). Examination of the stained cells revealed an association of anti-adipose IgG with cells exhibiting atypical morphology (dying or dead cells) (Fig. 4J,N). The serum dose-dependent increase in such atypical cells indicated that serum factors could cooperate with anti-adipose IgG to induce cell death, a classic feature of the complement system.

[0130] Example 7 Complement activation assay. 3T3-L1 cells were induced to differentiate under serum-free conditions in 96-well plates and allowed to accumulate lipids for up to 8 days. Cells were then preincubated in serum-free medium without or with antibodies for 40 minutes in a total volume of 60 μl, after which 20 μl of various dilutions of rabbit complement (BAG, Lich, Germany) in serum-free medium were added. To terminate the reaction, 40 μl of FluoroQuench AO / EB reagent (OneLambda, Canoga Park, USA) was added. After 15 minutes, fluorescent images were captured using a FITC filter (long-pass emission). Complement dilutions were freshly prepared and checked for activity before each individual experiment.

[0131] Example 8 Anti-fat autoantibodies induce complement-dependent cell death in 3T3-L1 cells. We examined whether anti-adipose IgG directs complement-mediated responses toward intact adipocytes. Complement alone or in the presence of control IgG had no effect on staining of dead or viable cells, respectively (Fig. 5). In contrast, within 1 h, anti-adipose IgG induced a significant increase in the number of dead cells in the presence of complement (Fig. 5A, B). Higher complement concentrations resulted in a further increase in the number of dead cells, reflected by a significant loss of viable cells (Fig. 5C). Extending the incubation time to 24 h had no effect on cell viability or the number of dead cells in the absence of anti-adipose IgG, but induced an even more pronounced loss of viable cells in the presence of anti-adipose IgG (Fig. 8). Apparently, the low-density epitope targeted by anti-perilipin-1 autoantibodies on the adipocyte plasma membrane (see Fig. 1D) was sufficient to induce complement-mediated cell death. Interestingly, after treatment with complement plus anti-adipose IgG, the remaining surviving cell population consisted primarily of preadipocytes lacking LSD and perilipin-1, again indicating the requirement for perilipin-1 expression in target cells for the anti-adipose IgG-induced complement response.

[0132] Example 9 Anti-perilipin-1 autoantibodies induce complement-mediated destruction of adipocytes in acquired systemic lipodystrophy. Systemic lipodystrophy is associated with severe metabolic complications. Familial forms have been identified as being due to defects in genes key to adipocyte function. The pathogenesis of acquired lipodystrophy is poorly understood, but it is often associated with chronic infection or autoimmune markers. As described in previous examples, we isolated autoantibodies against adipose tissue from the serum of patients with AGL. The immunoglobulin G fraction of patient serum, but not normal serum, specifically stained lipid-accumulating adipocytes in tissue sections and in vitro, indicating a direct immune response against adipocytes involved in the pathogenesis of AGL. Lipodystrophy-associated autoantibodies reacted with an intracellular autoantigen expressed exclusively in adipocytes and located on the surface of lipid storage droplets. This antigen was identified as perilipin-1 by two-dimensional electrophoresis coupled with mass spectrometry. The autoantibodies did not alter cellular lipid homeostasis or enter intact cells. However, in the presence of serum factors or complement, the autoantibodies induced cell death in cultured adipocytes. The present invention discloses a cellular target, namely perilipin-1, that plays a key role in the selective loss of adipocytes in AGL via a complement-mediated response.

[0133] We demonstrate that lipodystrophy-associated autoantibodies react with an autoantigen identified as perilipin-1, which is expressed exclusively in adipocytes and located on the surface of lipid storage droplets. The autoantibodies do not alter cellular lipid homeostasis or enter intact cells. However, in the presence of serum factors or complement, the autoantibodies induce cell death in cultured adipocytes. Anti-perilipin-1 autoantibodies are responsible for targeting the complement response to adipocytes.

[0134] Example 10 Antibody preparation. A naive human antibody-phage library was used for antibody selection of perilipin-1-specific antibodies. First, nonspecific or cross-reactive antibody-phages were removed from the library (campaign a). To this end, the library was incubated in the presence of immobilized streptavidin, magnetic streptavidin beads, and BSA. Antibody-phages that bound to negative antigens were removed from further selection.

[0135] The depleted library was then screened for target antigen-specific antibodies. The following peptide antigens were used for antibody selection: Perilipin epitope 1: NKGLTLLDGDLPEQE (SEQ ID NO: 330) Perilipin epitope 2: QYPPEKIASELKDTI (SEQ ID NO: 331) Perilipin epitope 5: AKPSLLSRVGALTNT (SEQ ID NO: 332) Perilipin epitope 6: EVRVPWLHSLAAAQE (SEQ ID NO: 333)

[0136] A peptide with a biotin-PEG spacer and a GGSGGS linker (SEQ ID NO: 438) at the N-terminus was synthesized. Additionally, a peptide with a biotin-PEG spacer and a AAAAA linker (SEQ ID NO: 439) at the C-terminus was also synthesized. Both peptide variants were purified by HPLC.

[0137] First, the target peptide (with an N-terminal biotin) was added to the depleted library preparation. Magnetic streptavidin beads were used to capture antibody-phage bound to the biotinylated target antigen and retrieved from the solution. To remove nonspecific or weakly bound antibody-phage particles, the beads were washed multiple times with BSA solution (containing 0.05% Tween® 20) and PBS. Antigen-specific antibody phages were eluted from the beads by trypsin treatment and rescued by E. coli infection. After a short growth period, the bacteria were co-infected with M13K07 helper phage to induce antibody-phage amplification. The amplified phage were used for two further selection cycles as described above. An overview of the antibody selection strategy is shown below. Selection strategies were performed for perilipin epitopes 1, 2, 5, and 6. Each strategy was performed separately with lambda and kappa antibody libraries.

[0138] [Table 1]

[0139] To direct antibody-phage selection toward cell-binding antibodies, antibody-phages were used for selection on the cell surface after selection cycle 2 of campaign a (campaign b). For this purpose, amplified phages were incubated in the presence of HEK293 cells. Antibody-phages bound to the HEK cell surface were removed from further selection. The removed antibody-phages were then selected for target antigen-specific antibodies by incubation in live adipocytes. Cells were washed multiple times to remove nonspecific or weakly bound antibody-phage particles. Antigen-specific antibody phages were rescued by E. coli infection. An overview of the antibody selection strategy is shown below. Each strategy was performed separately with lambda and kappa antibody-phages.

[0140] [Table 2]

[0141] Example 11 Antibody Screening After three rounds of antibody selection, the binding characteristics of the monoclonal antibody clones were analyzed. First, 384 single colonies from each perilipin selection strategy (perilipin epitopes 1, 2, 5, and 6) were used for scFv antibody production (campaigns a and b, respectively). These scFv productions were tested for their binding specificity by ELISA in the following: Streptavidin + target peptide (N-terminal biotin), Streptavidin + target peptide (C-terminal biotin), Streptavidin.

[0142] An antibody clone was identified as antigen-specific if: The signal / noise (S / N) between streptavidin + target peptide (N-terminal biotin) and streptavidin was ≥ 5, i.e., (streptavidin + target peptide (N-terminal biotin)) / (streptavidin) ≥ 5; The S / N ratio between streptavidin+target peptide (C-terminal biotin) and streptavidin is ≧5, ie, (streptavidin+target peptide (C-terminal biotin)) / (streptavidin)≧5.

[0143] A total of 827 hits were identified from campaign a. A total of 146 hits were identified from campaign b. Sequence analysis of these clones was performed to identify clones with unique antibody sequences (≥ 1 AA difference in the CDRs of VH and VL). An overview of the unique antibodies is shown below:

[0144] [Table 3]

[0145] Campaign b identified 18 antibodies that were identified in parallel in campaign a:

[0146] [Table 4]

[0147] Based on antibody selection campaign, peptide specificity and sequence similarity, lead antibodies were selected for further testing.

[0148] Example 12 Antibody conversion. VH and VL genes were amplified from phagemid DNA by PCR and purified. The VH gene was inserted into a mammalian human IgG1 heavy chain expression vector. The VL gene was inserted into a mammalian human IgG1 light chain lambda or kappa expression vector. The IgG expression vector was transformed into E. coli. Single colonies were selected for plasmid expansion. After isolation of transfection-grade DNA, HEK cells were transiently co-transfected with the heavy and light chain expression vector DNA of one antibody clone.

[0149] Antibodies were produced by HEK cells and secreted into the culture medium for 7 days. After clearance of the culture supernatant from the cells by centrifugation, the IgG antibodies were purified by protein A affinity chromatography. After adjusting the buffer to PBS, the antibody protein concentration was determined by UV / VIS spectrometry. The antibody integrity and purity were assessed by SDS-PAGE under reducing conditions. The functional binding activity of the antibodies to the target peptide was measured by ELISA. Various antibodies were produced and their functional characteristics were examined. In particular, the binding and functional characteristics of antibodies with various VH and VL regions were examined (SEQ ID NOs: 334-437).

[0150] Example 13 Exemplary anti-perilipin-1 antibodies of the invention. Various antibodies of the invention were tested in binding and functional assays, particularly hIgG1 antibodies raised against perilipin-1 epitopes having any of SEQ ID NOS: 1 (perilipin epitope 1), 2 (perilipin epitope 2), 5 (perilipin epitope 5), and 6 (perilipin epitope 6), respectively, and in particular any of SEQ ID NOS: 330, 331, 332, and 333.

[0151] [Table 5-1] [Table 5-2]

[0152] The respective sequences are detailed in the sequence listing and in Figures 16 (CDR sequences) and 17 (VH and VL sequences).

[0153] Example 14 Anti-perilipin antibody binding assay. Freshly isolated adipocytes from WT untreated mice were stained with selected antibodies. Specifically, adipocytes were isolated by collagenase digestion of adipose tissue (white fat from around the liver, intestine, kidney, and testis) from aged male mice [7, 8]. Collagenase digestion was performed using a digestion buffer (Krebs-Ringer-HEPES; 2.5 mM glucose; 2% fetal bovine serum albumin; 200 μM adenosine (Sigma, 01890-5G); 1 mg / ml collagenase (Sigma, C2139), pH 7.4, where Krebs-Ringer-HEPES contained 120 mM NaCl, 4.7 mM KCl, 2.2 mM CaCl2, 10 mM HEPES, 1.2 mM KH2PO4, and 1.2 mM MgSO4). Isolated adipocytes were first blocked with mouse FcR blocking reagent. Next, adipocytes were incubated with the selected primary antibody (anti-perilipin antibody or control) at a concentration of 1 μg / ml. As a control, an isotype control antibody (human IgG1) was used to exclude nonspecific antibody binding. Additionally, one control sample was incubated with only the secondary antibody without the addition of the primary antibody. After washing, adipocytes were incubated with a secondary antibody (goat anti-human Alexa 568) that recognized the primary antibody. After incubation and an additional washing step, DNA stain Hoechst 33342 was added at a 1:1000 dilution to identify intact cells by flow cytometry. Next, the binding of the anti-perilipin antibody to adipocytes was analyzed using flow cytometry.

[0154] For signal detection, anti-human Alexa 568, which recognizes the primary antibody, was used (Figures 10 and 11). Nonspecific binding of the tested antibodies to the cell surface was excluded using an isotype control antibody (human IgG1), i.e., by analyzing the fluorescence intensity of the anti-perilipin antibody compared to the isotype control. Histograms of the tested antibodies of the present invention showed enhanced binding of the antibodies of the present invention to adipocytes compared to the binding of the isotype control to adipocytes. Furthermore, the antibodies of the present invention showed enhanced binding to adipocytes compared to a commercially available anti-perilipin antibody (Figures 10 and 11).

[0155] Various antibodies of the present invention were tested for their binding efficiency on the cell surface of adipocytes. The binding of the antibodies of the present invention was further analyzed using gold nanoparticles and electron microscopy, essentially following a published protocol [9] with the modifications described below. Differentiated adipocytes on coverslips were fixed with 4% formaldehyde and 1% glutaraldehyde in PBS, pH 7.4, for 30 minutes. The fixed cell layer was then incubated overnight at 4°C without primary antibody or with 1 μg of each tested antibody per coverslip. After washing, the coverslips were incubated for 5 hours at 4°C with a crosslinking antibody: rabbit anti-human IgG (DAKO; catalog no. A0423; for any human or humanized primary antibody) or polyclonal mouse anti-rabbit (H+L) (Invitrogen; catalog no. 31213; for the commercially available rabbit anti-perilipin-1 polyclonal antibody PA1-1052 from Invitrogen). After washing, the coverslips were incubated overnight at 4°C with gold-labeled antibodies (Britisch Biocell International; goat anti-rabbit Au-20nm; catalog number EMGAR20; for human or humanized primary antibodies; or goat anti-mouse Au-20nm; catalog number EMGAM20; for commercially available primary antibodies). After a final wash, intact adipocytes were identified by their lipid vacuoles and microvilli, and bound gold particles were detected using field-emission scanning electron microscopy.

[0156] Successful binding of the antibodies of the present invention to adipocytes was observed. Binding of an antibody of the present invention, e.g., antibody 5, to adipocytes is shown in Figure 15. Similar results were observed for other antibodies of the present invention, e.g., antibodies 11, 17, 20, 29, and 31 (data not shown). In contrast, electron microscopy did not demonstrate binding of the commercially available antibody Invitrogen PA1-1052 to adipocytes.

[0157] Example 15 Anti-perilipin antibody functional assay. The effect of anti-perilipin antibodies on complement-mediated cell death was examined in a microscopy-based cell assay. 3T3-L1 cells were differentiated into adipocytes using an insulin-based serum-free differentiation protocol [6]. Specifically, 3T3-L1 cells were incubated for 3 days with serum-free differentiation medium (SFD medium) supplemented with insulin (1 μM), dexamethasone (0.1 μM), and 3-isobutyl-1-methylxanthine (500 μM). SFD medium contained DMEM / HAM's F12 (1:1) medium containing 15 mM HEPES mixed 1:1 with DMEM (1 g / L glucose without NaHCO3), 1.2 g / L NaHCO3, 17 μM D-pantothenic acid, 1 μM biotin, 2.0 mg / L apo-transferrin, 300 mg / L fetuin, 1% penicillin / streptomycin, pH 7.2. Cells were maintained in SFD medium supplemented with insulin (1 μM) only for 4 days. For complement assays

[10] , cells were incubated with selected antibodies (anti-perilipin and isotype control antibodies) at final concentrations between 0.017 and 0.05 mg / ml in 60 μl of SFD medium (pooled antibodies were each at a concentration of 0.0017 mg / ml per antibody in 60 μl of SFD medium). Commercially available lyophilized rabbit complement (BAG Diagnostics GmbH) was reconstituted in water, and then 20 μl of a 1:32 dilution of complement was added onto the cells. After 1 hour of incubation, cells were stained with FluoroQuench™ AE, and cell death was monitored using microscopic analysis.

[0158] Successful differentiation was assessed based on morphological changes in the cells, such as the appearance of lipid vacuoles and cell enlargement. Differentiated cells were incubated with selected antibodies and complement. Cell death was monitored using microscopic analysis and FluoroQuench™ staining / quenching reagents commercially available from Thermo Fisher Scientific Inc.

[0159] Antibodies of the invention demonstrated efficient complement-mediated cell death (Figures 12B and 13B). Various antibodies of the invention were tested for their functional efficacy and demonstrated effective complement-mediated cell death (Figure 12B), e.g., antibodies 6, 7, 8, 9, 14, 16, 18, 19, 20, 21, 22, 24, 25, 26, 27, 28, 31, and 32. Commercially available anti-perilipin antibodies tested did not demonstrate significant complement-mediated cell death compared to controls (Figures 12A, 14, and 15C).

[0160] Example 16 Further evaluation of exemplary antibodies of the invention. Various antibodies of the present invention were further investigated for their adipocyte binding efficacy and their efficacy in complement-mediated cell death (Table 6).

[0161] [Table 6]

[0162] Furthermore, complementation assays were performed with antibody pools to investigate the synergistic effect of targeting several perilipin epitopes (Table 7).

[0163] [Table 7]

[0164] References [1]Greenberg AS, Egan JJ, Wek SA, Moos MC, Jr., Londos C, Kimmel AR. Isolation of cDNAs for perilipins A and B: sequence and expression of lipid droplet associated proteins of adipocytes. Proc Natl Acad Sci USA, 1993;90:12035-9. [2]Huebler A, Abendroth K, Keiner T, Storcker W, Kauf E, Hein G et al. Dysregulation of insulin-like growth factors in a case of generalized acquired lipoatrophic diabetes mellitus (Lawrence Syndrome) connected with autoantibodies against adipocyte membranes. Exp Clin Endocrinol Diabetes, 1998;106:79-84. [3]Corvillo F, Aparicio V, Lopez-Lera A, Garrido S, Araujo-Vilar D, et al. Autoantibodies against perilipin 1 as a cause of acquired generalized lipodystrophy. Front Immunol, 2018; 9:2142. [4] Misra A, Garg A. Clinical features and metabolic derangements in acquired generalized lipodystrophy: case reports and review of the literature. Medicine (Baltimore), 2003;82:129-46. [5]Schmidt M, Lorffler G. Induction of aromatase activity in human adipose tissue stromal cells by extracellular nucleotides--evidence for P2-purinoceptors in adipose tissue. Eur J Biochem, 1998;252:147-54. [6]Schmidt W, Poell-Jordan G, Lorffler G. Adipose conversion of 3T3-L1 cells in a serum-free culture system depends on epidermal growth factor, insulin-like growth factor I, corticosterone, and cyclic AMP. J Biol Chem, 1990;265:15489-95. [7]Rodbell M. Metabolism of isolated fat cells. I. Effects of hormones on glucose metabolism and lipolysis. J Biol Chem, 1964;239:375-80. [8]Schordel J, Weise I, Klinger R, Schmidt M. Stimulation of lipogenesis in rat adipocytes by ATP, a ligand for P2-receptors. Biochem Biophys Res Commun, 2004; 321:767-73. [9]Kaufmann R, Rahn S, Pollrich K, Hertel J, Dittmar Y, Hommann M, Henklein P, Biskup C, Westermann M, Hollenberg MD, Settmacher U. Thrombin-mediated hepatocellular carcinoma cell migration: cooperative action via proteinase-activated receptors 1 and 4. J Cell Physiol, 2007; 211:699-707.

[10] Terasaki PI, McClelland JD. Microdroplet assay of human serum cytotoxins. Nature, 1964;204:998-1000.

[0165] The features of the invention disclosed in this specification, the claims and / or the accompanying drawings, both separately and in any combination thereof, may be material for realizing the invention in various of their forms. The present invention provides, for example, the following items. (Item 1) An antigen-binding peptide that specifically binds to at least one perilipin-1 epitope selected from the following amino acid sequences: LTLLDGDLPE (SEQ ID NO: 1), EKIASELK (SEQ ID NO: 2), VPIASTSDKV (SEQ ID NO: 3), PAPGHQQAQK (SEQ ID NO: 4), PSLLSRVGALTN (SEQ ID NO: 5), and PWLHSLAA (SEQ ID NO: 6). (Item 2) 2. The antigen-binding peptide according to item 1, which binds to two or more of SEQ ID NOs: 1 to 6. (Item 3) 3. The antigen-binding peptide according to item 1 or 2, wherein the perilipin-1 epitope is a human perilipin-1 epitope. (Item 4) 10. The antigen-binding peptide of any of the preceding items, which is not a perilipin-1-targeting autoantibody in a human patient with lipoatrophy, and preferably is not a perilipin-1-targeting anti-PLIN1 antibody against a peptide spanning Thr8 to Ala145 having manufacturer R&D catalog #AF6615. (Item 5) 10. The antigen-binding peptide of any of the preceding items, which competes for binding with perilipin-1-targeting autoantibodies of human patients with lipoatrophy. (Item 6) In the binding assay, at least 10 -4 Affinity of M (K D 10. The antigen-binding peptide of any preceding item, which binds to perilipin-1 at (Item 7) 10. The antigen-binding peptide of any of the preceding items, which is an antibody, preferably a monoclonal antibody. (Item 8) 2. The antigen-binding peptide of any of the preceding items, wherein, when the antigen-binding peptide, particularly the antibody, is incubated with 3T3-L1 cells differentiated into adipocytes in the presence of complement, it causes complement-mediated cell death of the 3T3-L1 cells and / or the differentiated adipocytes. (Item 9) 10. The antigen-binding peptide of any of the preceding items, wherein when the antigen-binding peptide, particularly the antibody, is incubated with adipocytes, the antigen-binding peptide, particularly the antibody, binds to the surface of the adipocytes. (Item 10) i. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 18, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 20, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 22, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 23, or ii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 24, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 25, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 28, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 29, or iii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 30, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 31, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 32, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 34, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 35, or iv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 36, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 37, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 38, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 40, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 41, or v. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 42, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 43, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 44, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 45, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 46, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 47, or vi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 48, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 49, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 50, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 51, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 52, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 53, or vii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 54, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 55, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 56, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 57, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 58, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 59; or viii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 60, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 61, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 62, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 63, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 64, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 65; or ix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 66, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 67, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 68, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 69, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 70, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 71; or x. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 72, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 73, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 74, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 75, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 76, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 77, or xi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 78, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 79, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 80, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 81, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 82, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 83; or xii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 84, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 85, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 86, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 87, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 88, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 89, or xiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 90, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 91, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 92, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 93, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 94, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 95, or xiv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 96, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 97, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 98, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 99, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 100, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 101, or xv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 102, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 103, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 104, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 105, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 106, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 107, or xvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 108, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 109, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 110, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 111, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 112, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 113; or xvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 114, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 115, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 116, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 117, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 118, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 119; or xviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 120, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 121, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 122, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 123, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 124, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 125; or xix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 126, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 127, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 128, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 129, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 130, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 131; or xx. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 132, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 133, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 134, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 135, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 136, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 137, or xxi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 138, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 139, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 140, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 141, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 142, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 143; or xxii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 144, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 145, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 146, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 147, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 148, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 149; or xxiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 150, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 151, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 152, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 153, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 154, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 155; or xxiv. A heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 156, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 157, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 158, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 159, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 160, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 161, or xxv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 162, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 163, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 164, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 165, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 166, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 167; or xxvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 168, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 169, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 170, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 171, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 172, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 173; or xxvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 174, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 175, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 176, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 177, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 178, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 179; or xxviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 180, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 181, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 182, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 183, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 184, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 185; or xxix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 186, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 187, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 188, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 189, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 190, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 191; or xxx. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 192, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 193, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 194, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 195, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 196, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 197; or xxxi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 198, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 199, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 200, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 201, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 202, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 203; or xxxii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 204, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 205, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 206, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 207, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 208, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 209; or xxxiii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 210, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 211, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 212, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 213, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 214, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 215; or xxxiv. A heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 216, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 217, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 218, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 219, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 220, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 221, or xxxv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 222, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 223, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 224, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 225, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 226, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 227; or xxxvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 228, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 229, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 230, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 231, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 232, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 233; or xxxvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 234, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 235, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 236, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 237, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 238, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 239; or xxxviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 240, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 241, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 242, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 243, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 244, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 245; or xxxix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 246, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 247, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 248, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 249, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 250, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 251; or xl. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 252, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 253, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 254, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 255, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 256, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 257; or xli. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 258, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 259, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 260, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 261, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 262, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 263; or xlii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 264, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 265, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 266, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 267, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 268, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 269; or xliii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 270, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 271, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 272, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 273, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 274, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 275; or xliv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 276, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 277, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 278, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 279, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 280, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 281; or xlv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 282, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 283, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 284, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 285, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 286, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 287, or xlvi. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 288, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 289, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 290, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 291, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 292, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 293; or xlvii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 294, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 295, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 296, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 297, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 298, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 299; or xlviii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 300, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 301, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 302, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 303, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 304, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 305; or xlix. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 306, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 307, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 308, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 309, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 310, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 311; or l. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 312, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 313, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 314, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 315, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 316, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 317; or a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 318, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 319, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 320, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 321, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 322, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 323; or lii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 324, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 325, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 326, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 327, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 328, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 329; an antigen-binding peptide, preferably an antibody; Alternatively, a peptide, the heavy chain CDR1, CDR2, CDR3 regions of which and the light chain CDR1, CDR2, CDR3 regions of which comprise amino acid sequences having at least 90% identity, preferably at least 95% identity, more preferably at least 98% identity, even more preferably 99% identity to any of the sequences set out in i. to lii. or the antigen-binding peptide of any of the preceding items, which is an antibody. (Item 11) 10. The antigen-binding peptide of any of the preceding items, which is a human antibody, a humanized antibody, or a chimeric human antibody. (Item 12) Intact antibody, substantially intact antibody, Fab fragment, F(ab') 2 10. The antigen-binding peptide of any of the preceding items, which is a fragment or a single-chain Fv fragment. (Item 13) 10. The antigen-binding peptide of any of the preceding items, capable of binding to a perilipin-1 fragment displayed on an adipocyte, wherein the binding of the antigen-binding peptide to the perilipin-1 fragment on the surface of the adipocyte activates the immune effector system to target and / or eliminate the adipocyte. (Item 14) 10. The antigen-binding peptide of any of the preceding items, comprising an Fc region, preferably an Fc region that has been optimized by protein engineering for higher binding affinity to effector cells compared to the binding affinity of a non-optimized Fc region. (Item 15) The antigen-binding peptide does not cross-react with any other perilipin proteins, such as perilipin-2, -3, -4, or -5. (Item 16) 10. The antigen-binding peptide of any of the preceding items for use in the prevention or treatment of obesity. (Item 17) (i) a first binding site that specifically binds to a perilipin-1 epitope having any of SEQ ID NOs: 1 to 6; (ii) a second binding site; and A bispecific antigen-binding peptide comprising: (Item 18) Item 18. The bispecific antigen-binding peptide according to Item 17, wherein the second binding site binds to a perilipin-1 epitope having any of SEQ ID NOs: 1 to 6 that is identical to or different from the epitope of the first binding site; to a white adipocyte-specific surface marker, for example, the neutral amino acid transporter ASC-1; or to an immune cell-specific receptor molecule, such as a T cell-specific receptor molecule or a natural killer cell-specific receptor molecule. (Item 19) 19. The bispecific antigen-binding peptide of item 17 or 18 for use in the prevention or treatment of obesity. (Item 20) A composition comprising the antigen-binding peptide according to any one of items 1 to 15 and / or the bispecific antigen-binding peptide according to any one of items 17 to 18. (Item 21) 21. The composition according to item 20, comprising two or more antigen-binding peptides according to any one of items 1 to 15 and / or two or more bispecific antigen-binding peptides according to any one of items 17 to 18. (Item 22) 22. The composition of claim 21, wherein the two or more antigen-binding peptides are a set of peptides, each of which binds to a different perilipin-1 epitope, and the two or more bispecific antigen-binding peptides are a set of bispecific peptides, each of which binds to a different perilipin-1 epitope. (Item 23) 16. The composition according to claim 1, wherein the at least two or at least three or at least four, or at least five, or at least six antigen-binding peptides, each of which binds to a different perilipin-1 epitope; and / or the composition according to any of items 17-18, wherein the composition comprises at least two, or at least three, or at least four, or at least five, or at least six bispecific antigen-binding peptides, each of which binds to a different perilipin-1 epitope. (Item 24) 24. The composition according to any of items 20 to 23 for use as a pharmaceutical composition in the prevention or treatment of obesity. (Item 25) A pharmaceutical composition comprising the antigen-binding peptide according to any one of items 1 to 15 and / or the bispecific antigen-binding peptide according to any one of items 17 to 18 for use in the prevention or treatment of obesity. (Item 26) An isolated nucleic acid encoding the antigen-binding peptide of any one of Items 1 to 15, or encoding the bispecific antigen-binding peptide of Item 17 or 18. (Item 27) 27. A recombinant cell comprising the nucleic acid of item 26, which is preferably a recombinant autologous effector cell or a recombinant host cell. (Item 28) 1. A method for producing a means for specifically targeting perilipin-1 fragment-presenting adipocytes, comprising: a) inoculating a non-human animal with an antigen comprising at least one perilipin-1-specific epitope selected from SEQ ID NOs: 1 to 6, and screening and producing antigen-binding peptides that specifically bind to said at least one perilipin-1-specific epitope from said inoculated animal, preferably by collecting cells expressing said antigen-binding peptide from the spleen of said non-human animal, preparing hybridoma cells therefrom, expressing said antigen-binding peptide, and screening the antigen-binding peptides expressed by said hybridoma cells to identify antigen-binding peptides that specifically bind to any of SEQ ID NOs: 1 to 6 and isolates thereof; or b) providing and screening a recombinant antigen-binding peptide library, such as a library of antibodies, scFv molecules, camelid antibodies, anticalins or DARPins, to identify antigen-binding peptides that specifically bind to any of SEQ ID NOS: 1-6 and isolates thereof; or c) providing and screening a cell pool containing cytotoxic T cells, such as by tetramer staining or dextramers techniques, to identify cytotoxic T cells having specificity for any of SEQ ID NOS: 1-6 and isolates thereof; or d) providing a patient's blood sample and obtaining T cells therefrom, and genetically modifying said T cells by introducing the nucleic acid according to item 18, and obtaining chimeric antigen receptor-expressing T cells; or e) providing and screening a pool of T cell receptors to identify T cell receptors having specificity for any of SEQ ID NOS: 1-6 and isolates thereof; A method comprising: (Item 29) 29. The method of claim 28, wherein the means targets perilipin-1 and is selected from an antigen-binding peptide and a cytotoxic T cell.

Claims

1. 1. An antigen-binding peptide that specifically binds to a perilipin-1 epitope having the following amino acid sequence: LTLLDGDLPE (SEQ ID NO: 1), the antigen-binding peptide i. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 18, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 19, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 20, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 22, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 23, or ii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 24, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 25, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 27, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 28, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 29, or iii. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 30, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 31, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 32, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 33, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 34, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 35, or iv. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 36, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 37, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 38, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 39, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 40, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 41, or v. comprising a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 42, a heavy chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 43, a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 44, a light chain CDR1 region comprising the amino acid sequence of SEQ ID NO: 45, a light chain CDR2 region comprising the amino acid sequence of SEQ ID NO: 46, and a light chain CDR3 region comprising the amino acid sequence of SEQ ID NO: 47; An antigen-binding peptide that is an antibody or antibody fragment.

2. The antigen-binding peptide of claim 1, wherein the perilipin-1 epitope is a human perilipin-1 epitope.

3. The antigen-binding peptide of any one of claims 1 to 2, which is not a perilipin-1-targeting autoantibody in a human patient with lipoatrophy.

4. 4. The antigen-binding peptide of any one of claims 1 to 3, which competes for binding with perilipin-1-targeting autoantibodies of human patients with lipoatrophy.

5. In the binding assay, at least 10 －４ Affinity of M (K Ｄ The antigen-binding peptide of any one of claims 1 to 4, which binds to perilipin-1 at the nucleotide sequence (SEQ ID NO: 1).

6. The antigen-binding peptide of any one of claims 1 to 5, which is a monoclonal antibody.

7. The antigen-binding peptide of any one of claims 1 to 6, wherein the antibody, when incubated with 3T3-L1 cells differentiated into adipocytes in the presence of complement, causes complement-mediated cell death of the 3T3-L1 cells and / or the differentiated adipocytes.

8. The antigen-binding peptide according to any one of claims 1 to 7, wherein the antibody binds to the surface of adipocytes when incubated with the adipocytes.

9. The antigen-binding peptide of any one of claims 1 to 8, which is a human antibody or a chimeric human antibody.

10. Intact antibody, Fab fragment, F(ab') ２ The antigen-binding peptide of any one of claims 1 to 9, which is a fragment or a single-chain Fv fragment.

11. 11. The antigen-binding peptide of any of claims 1 to 10, which is capable of binding to a perilipin-1 fragment presented on an adipocyte, and wherein the binding of the antigen-binding peptide to the perilipin-1 fragment on the adipocyte activates the immune effector system to target and / or eliminate the adipocyte.

12. 12. The antigen-binding peptide of any one of claims 1 to 11, comprising an Fc region that has been optimized by protein engineering for higher binding affinity to effector cells compared to the binding affinity of a non-optimized Fc region.

13. An antigen-binding peptide according to any one of claims 1 to 12, which does not cross-react with perilipin proteins other than perilipin-1 (e.g., perilipin-2, -3, -4 or -5).

14. The antigen-binding peptide of any one of claims 1 to 13 for use in the prevention or treatment of obesity.

15. (i) a first binding site that specifically binds to the perilipin-1 epitope having SEQ ID NO: 1, wherein the first binding site is an antibody or antibody fragment of claim 1; (ii) a second binding site; and A bispecific antigen-binding peptide comprising:

16. 16. The bispecific antigen-binding peptide of claim 15, wherein the second binding site binds to a perilipin-1 epitope having any of SEQ ID NOs: 1-6 that is identical to or different from the epitope of the first binding site; or to an immune cell-specific receptor molecule (e.g., a T cell-specific receptor molecule or a natural killer cell-specific receptor molecule).

17. 17. The bispecific antigen-binding peptide of claim 15 or 16 for use in the prevention or treatment of obesity.

18. A composition comprising the antigen-binding peptide of any one of claims 1 to 13 and / or the bispecific antigen-binding peptide of claim 15 or 16.

19. The composition of claim 18, comprising two or more antigen-binding peptides of any one of claims 1 to 13, and / or two or more bispecific antigen-binding peptides of claim 15 or 16.

20. 20. The composition of claim 18 or 19 for use as a pharmaceutical composition in the prevention or treatment of obesity.

21. A pharmaceutical composition comprising the antigen-binding peptide of any one of claims 1 to 13 and / or the bispecific antigen-binding peptide of claim 15 or 16 for use in the prevention or treatment of obesity.

22. An isolated nucleic acid encoding an antigen-binding peptide according to any one of claims 1 to 13, or encoding a bispecific antigen-binding peptide according to claim 15 or 16.

23. 23. A recombinant cell comprising the nucleic acid of claim 22.

24. 24. The recombinant cell of claim 23, which is a recombinant autologous effector cell or a recombinant host cell.

25. 1. A method for producing a means for specifically targeting perilipin-1 fragment-presenting adipocytes, comprising: a) inoculating a non-human animal with an antigen comprising a perilipin-1-specific epitope having SEQ ID NO: 1, and screening and producing antigen-binding peptides from the inoculated animal that specifically bind to the perilipin-1-specific epitope, wherein the antigen-binding peptides are antibodies or antibody fragments; or b) providing and screening a library of recombinant antigen-binding peptides (e.g., antibodies, scFv molecules, camelid antibodies, anticalins, or DARPins) to identify antigen-binding peptides that specifically bind to the perilipin-1-specific epitope having SEQ ID NO: 1; or c) screening a cell pool containing cytotoxic T cells to identify cytotoxic T cells having specificity for the perilipin-1-specific epitope having SEQ ID NO: 1; or d) obtaining T cells from a patient's blood sample, genetically modifying the T cells by introducing a nucleic acid encoding a chimeric antigen receptor comprising the antigen-binding peptide of claim 1, and obtaining chimeric antigen receptor-expressing T cells; or e) screening the pool of T cell receptors to identify T cell receptors with specificity for the perilipin-1 specific epitope having SEQ ID NO: 1 A method comprising:

26. Step a) comprises: a) inoculating a non-human animal with an antigen comprising a perilipin-1-specific epitope having SEQ ID NO: 1; and screening and producing antigen-binding peptides that specifically bind to the perilipin-1-specific epitope having SEQ ID NO: 1 from the inoculated animal by collecting cells expressing the antigen-binding peptide from the spleen of the non-human animal, preparing hybridoma cells therefrom, expressing the antigen-binding peptide, and screening the antigen-binding peptides expressed by the hybridoma cells to identify antigen-binding peptides that specifically bind to the perilipin-1-specific epitope having SEQ ID NO: 1, wherein the antigen-binding peptide is an antibody or an antibody fragment.

26. The method of claim 25, wherein the method is performed as follows:

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