Means for specifically eliminating perilipin-1 fragment presenting adipocytes

Abstract

To provide means for specifically eliminating perilipin-1 fragment presenting adipocytes.SOLUTION: The present invention relates to an antigen-binding peptide specifically binding to at least one perilipin-1 epitope as defined herein. The present invention further relates to an antigen-binding peptide for use in the prevention or treatment of obesity. Furthermore, the present invention relates to a bispecific antigen-binding peptide comprising a first binding site specifically binding to a perilipin-1 epitope and a second binding site, and use thereof. The present invention also relates to a pharmaceutical composition comprising an antigen-binding peptide or a bispecific antigen-binding peptide, for use in the prevention or treatment of obesity. The present invention further relates to an isolated nucleic acid encoding an antigen-binding peptide or a bispecific antigen-binding peptide.SELECTED DRAWING: None

Description

[Technical field]

[0001] FIELD OF THEINVENTION 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 said nucleic acids, and to methods of producing means for specifically targeting perilipin-1 fragment-presenting adipocytes. [Background technology]

[0002] 2. Background of the Invention Obesity is a global problem, and according to the World Health Organization, over 1.9 billion people are overweight. People suffering from obesity are generally susceptible to various medical conditions. Surgical intervention is currently a successful therapeutic approach, but complications often occur with subsequent gastrointestinal problems. Thus, surgical intervention is considered acceptable only in severely obese patients with high comorbidities. Relatively promising pharmacological therapies have had to be withdrawn from the market due to severe side effect profiles. The pleiotropic nature of the signaling mechanisms involved has led to unacceptable side effects. Thus, there is a lack of therapeutic approaches that are successful in 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 that result in the loss of body fat, namely lipodystrophy. Although metabolic complications vary with respect to the degree of fat loss, the common consequences of lipodystrophy are insulin resistance or diabetes mellitus, absence of ketosis, increased metabolic rate, hyperlipidemia, and hepatomegaly, including fatty liver. These consequences, as shown in various animal models, are due to 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 such other cells, other members of lipid droplet-associated proteins, such as adipophilin and TIP47, are present instead of perilipin-1. Since 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 result in increased lipolysis and release of lipids stored in the droplets (and thus exacerbating the metabolic situation). Thus, a reduction in fat mass is expected at the expense of an unfavorable metabolic situation, which can be prevented by completely eliminating the adipocytes. Since other types of lipid storage cells do not present perilipin-1, targeting perilipin-1 for activation of immune effector mechanisms against adipocytes can allow for the specific elimination of adipocytes. WO02 / 028410A1 discloses that perilipin may be a target for the regulation of 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 reactions against adipocytes result in almost 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 lipodystrophic patients can be used to specifically target adipose tissue in patients suffering from obesity.However, such a specific effector system, which can be, for example, an antibody or an antigen-binding fragment or a cytotoxic T cell, has not been obtained.In particular, despite great efforts, no adipocyte-specific epitope has been identified to date that can potentially be targeted as a specific therapeutic target [4]. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO 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, the present invention aims to provide a means for specifically targeting and / or eliminating adipocytes, such as antigen-binding peptides or cytotoxic T cells. Furthermore, the present invention aims 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 target and / or eliminate adipocytes by activating immune effector systems, such as extrinsic or endogenous effector systems. 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 from 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 said epitopes useful in the treatment of obesity. The present inventors have 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 such 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 diseases and / or medical conditions related to obesity.

[0013] The present invention aims at the use of exogenous or endogenous effector systems of the immune system to specifically target and / or eliminate adipocytes. Such exogenous effector systems can be, for example, ex vivo or in vitro generated and / or amplified antibodies and / or cytotoxic T cells. The epitopes of perilipin-1 disclosed herein, SEQ ID NOs: 1-6, 330-333, allow for specific targeting of adipocytes via perilipin-1 fragments presented on said adipocytes, in particular those present on the surface of said adipocytes. Targeting of 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, which leads to activation of FcRIII (CD16), Fc-RII (CD32), Fc-RI (CD64) or other complement system and / or Fc-receptors, which leads to recruitment and activation of effector systems such as ADCC (antibody-dependent cellular 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, in particular 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 directly eliminate the adipocytes in the case of ADCP, thus providing a direct means of elimination, or may indirectly eliminate the adipocytes by targeting them, resulting in activation of effector systems such as the complement system or ADCC or cytotoxic T cells, thus providing an indirect means of 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 to specifically target and / or eliminate adipocytes without off-target effects in other cells and thus with negligible side effects.

[0015] Furthermore, targeting of fat cells for the immune effector system 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 uses adipocyte-specific presentation on the surface of adipocytes of typically intracellular proteins, i.e., fragments of proteins previously only known to exist intracellularly, i.e., perilipin-1 fragments, which are essential for lipid storage in adipocytes. The present inventors herein disclose six peptide sequences that are epitopes in perilipin-1, identified by analyzing the binding patterns of anti-perilipin-1 autoantibodies from lipodystrophy patients. The present inventors also disclose further peptide sequences that are variants of such 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] By marking said adipocytes in vivo with antibodies, antibody fragments or other suitable affinity ligands produced by the method according to a) or b), it is possible to induce a specific immune effector response.Furthermore, adipocytes can be indirectly eliminated in vivo with cytotoxic T cells produced by the method according to c).

[0019] Hereinafter, the elements of the present invention are described. Although these elements are listed according to 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 interpreted as limiting the present invention to only the embodiments specifically described. This description should be understood as supporting and encompassing 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. Moreover, 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, said 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 of 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, in particular 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 vs. dead cells using a suitable dye or mixture of dyes, such as a fluorescent dye or mixture of fluorescent dyes. Typically, such dye or mixture of dyes distinguishes between live and dead cells. An example of such a mixture of dyes is the dye commercially available as "FluoroQuench™ Staining / Quenching Reagent" from ThermoFisher. 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 further supplemented with dexamethasone and / or isobutyl-1-methylxanthine as required.

[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, said antigen-binding peptide, in particular said antibody, when incubated with adipocytes, binds to the surface of said adipocytes. In one embodiment, said binding of said antigen-binding peptide, preferably said antibody, to the surface of said adipocytes is determined by incubating said 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 said 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 external surface of such adipocyte, i.e., the extracellular surface exposed to the surroundings of the adipocyte.

[0033] In one embodiment, the antigen-binding peptide, in particular 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 binds to the surface of the adipocytes after incubation with the adipocytes.

[0034] In one embodiment, the antigen-binding peptide comprises: 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. comprises 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. comprises 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. comprises 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. comprising 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. comprising 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, CDR3 regions and whose light chain CDR1, CDR2, CDR3 regions comprise 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.

[0035] In one embodiment, the antigen-binding peptide comprises: i. comprises the amino acid sequence of SEQ ID NO: 334 and / or comprises the amino acid sequence of SEQ ID NO: 335; or ii. comprises the amino acid sequence of SEQ ID NO: 336 and / or comprises the amino acid sequence of SEQ ID NO: 337; or iii. comprises the amino acid sequence of SEQ ID NO: 338 and / or comprises 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. comprises the amino acid sequence of SEQ ID NO: 346 and / or comprises the amino acid sequence of SEQ ID NO: 347; or viii. comprises the amino acid sequence of SEQ ID NO: 348 and / or comprises 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. comprises the amino acid sequence of SEQ ID NO: 356 and / or comprises the amino acid sequence of SEQ ID NO: 357; or xiii. comprises the amino acid sequence of SEQ ID NO: 358 and / or comprises 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. comprises the amino acid sequence of SEQ ID NO: 366 and / or comprises 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 li. 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 at least 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 SEQ ID NO:, is typically intended to allow for the presence of additional residues flanking such sequence, which 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 include 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" an 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 no additional flanking residues are 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 It is a single chain Fv fragment or a single chain Fv fragment.

[0041] In one embodiment, the antigen-binding peptide is capable of binding to a perilipin-1 fragment presented 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, said antigen-binding peptide is an antigen-binding peptide for use in the prevention or treatment of obesity.

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

[0046] In a further aspect, the present invention provides a method for producing a method for treating a cancer cell comprising the steps of: (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 that 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, said 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, a 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] Said antigen-binding peptide and said 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] Said antigen-binding peptide and said 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: 1-6, 330-333, 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 harvesting cells expressing an antigen-binding peptide from the spleen of said non-human animal, preparing hybridoma cells therefrom, expressing said antigen-binding peptide, and screening said antigen-binding peptides expressed by said hybridoma cells to identify antigen-binding peptides that specifically bind to any of SEQ ID NOs: 1-6, 330-333, 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 pool of cells containing cytotoxic T cells, such as by tetramer staining or dextramer 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 the steps of:

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

[0063] Said antigen-binding peptide and said 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] Said antigen-binding peptide, said bispecific antigen-binding peptide and said 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 PREFERRED EMBODIMENTS

[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 to 6. In one embodiment, said perilipin-1 epitope is selected from any of SEQ ID NOs: 1 to 6, 330 to 333. In one embodiment, the antigen-binding molecule is based on an immunoglobulin, such as a polyclonal or monoclonal antibody, or is an anticalin protein, an affilin, an affimer, an affitin, an alphabody, a nanobody or a D-cell antigen. Based on protein scaffold structures with antigen-binding capacity, such as ARPin. In one embodiment, the antigen-binding peptide relates to an antibody or antibody fragment. In one embodiment, when referring to an antibody or antibody fragment, reference is made 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 mainly by plasma cells of the immune system to neutralize pathogens. Antibodies are grouped into classes, also called isotypes, which are genetically determined by the constant region. 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, which define the isotype of the antibody as IgM, IgD, IgG, IgA and IgE, respectively. The IgG class is the most commonly used for therapeutic purposes. "IgG" as used herein refers to a polypeptide belonging to the class of antibodies substantially encoded by the recognized immunoglobulin gamma genes. In humans, this class includes the subclasses IgG1, IgG2, IgG3 and IgG4. In mice, this class includes the 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 classes or subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions. The known human immunoglobulin isotypes are IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgM, IgD and IgE. IgG, which is a hybrid composition of the 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 between 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 a domain of 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 provide 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, human antibodies, human chimeric antibodies, humanized antibodies, and antibody fragments. The antibody recognizes an antigen via a Fab fragment variable region that contains 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 having SEQ ID NOs: 1-6, 330-333 in perilipin-1. In one embodiment, the binding of the antigen-binding peptide to a perilipin-1 fragment presented 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 a bispecific molecule, perilipin-1 and, optionally, another antigen, such as an antigen on the surface of an effector cell, such as an antigen on the surface of a cytotoxic T cell, or an antigen presented on the surface of an adipocyte 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 and relates 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, said 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, said TCR-derived antigen-binding peptide is used for adoptive T cell receptor transfer. In one embodiment, the antigen-binding peptide relates to a peptide expressed on the surface of a genetically modified cell, in particular a T cell or an NK cell, said genetically modified cell being genetically modified to express said antigen-binding peptide, resulting in a CAR-T cell or a CAR-NK cell, respectively.

[0076] In the present invention, the antigen-binding peptide further includes a substantially intact antibody, a Fab fragment, a F(ab') 2The antigen-binding peptide may relate to a fragment of an antibody, such as a fragment, a diabody, a single chain Fv fragment, a tetrabody, a triabody, a disulfide bond stabilized Fv (dsFv) or a heavy chain VHH fragment from camelid. However, other forms of antigen-binding peptides are also envisaged 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, for example by using the method of combinatorial protein design. Such a non-antibody antigen-binding peptide may for example be an affibody molecule, an affilin, an affimer, an affitin, an alphabody, an anticalin, a nanobody or a DARPin. The antigen-binding peptide may also relate to other antibody mimetic molecules, such as a dual-affinity re-targeting antibody (DART). In one embodiment, the antigen-binding peptide preferably relates to an antibody or an antibody fragment. In one embodiment, the antigen-binding peptide can refer to any antibody mimetic molecule that mimics the binding properties of an antibody to an antigen, but is structurally distinct from a naturally occurring antibody.

[0077] In one embodiment, the antigen-binding peptide can be optimized by protein engineering methods known to those skilled in the art, such as directed evolution, phage display, or ribosome display, to have improved properties, such as higher binding affinity and longer serum half-life, compared to non-optimized antigen-binding peptides. In one embodiment, the antigen-binding peptide can be modified by protein engineering methods known to those skilled in the art to have the ability to induce certain effector functions, such as by including an Fc region to be targeted by effector cells. In one embodiment, the optimized antigen-binding peptide can be engineered to increase the response of effector systems to the Fc region by exchange of 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 improved serum half-life, for example, by modification of 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 to improve its ability to induce a certain effector function, such as a stronger effector response by the complement system or cytotoxic T cells, when it recognizes the optimized antigen-binding peptide bound to an epitope having any of SEQ ID NOs: 1-6, 330-333 presented on or present on the surface of adipocytes, compared to a non-optimized antigen-binding peptide bound to an epitope having any of SEQ ID NOs: 1-6, 330-333 presented on or present on the surface of adipocytes. 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 of the 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 contribute together with the CDRs from the other chain to form the antigen binding site. In one embodiment, the antigen binding peptide comprises at least one, preferably all, CDRs of an antibody that specifically binds to a perilipin-1 fragment, preferably specifically binds 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, preferably all, CDRs of an antibody that specifically binds to a perilipin-1 fragment, said peptide scaffold being an antibody fragment or any non-antibody peptide scaffold, such as anticalin, affilin, affimer, affitin, alphabody, nanobody or DARPin.

[0079] The term "bispecific antigen-binding peptide" as used herein relates 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. A bispecific antigen-binding peptide can simultaneously bind to two target structures, such as to two different types of antigens. In one embodiment, the bispecific antigen-binding peptide is based on an immunoglobulin. In one embodiment, the bispecific antigen-binding peptide comprises binding fragments based on two different immunoglobulins with binding specificity to an antigen, the two fragments being connected by a linker fragment, such as a linker peptide sequence, and / or an antibody scaffold structure. In one embodiment, the binding fragments of a bispecific antigen-binding peptide can be specific for different antigens, or for different epitopes on the same antigen, or for the same epitope on the same antigen. In one embodiment, a bispecific antigen-binding peptide according to the invention binds to a first perilipin-1 epitope and a second, but different, non-overlapping perilipin-1 epitope. In one embodiment, the bispecific antigen-binding peptide of the present invention binds to a perilipin-1 epitope and to a different antigen. In one embodiment, the different antigen is a surface marker of an effector cell, such as a T cell or a natural killer cell. In one embodiment, the different antigen may be an antigen presented on the surface of an adipocyte 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, anticalin, DARPin, aptamer, affibody molecule, affilin, affimer, affitin, alphabody, and nanobody.In one embodiment, the second binding site specifically binds to a receptor on an immune cell, such as a T cell or a natural killer cell, preferably a receptor capable of activating the immune cell or stimulating 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, CD3 delta / epsilon, CD3 gamma / epsilon, TCR, TCR alpha, TCR beta, CD2, CD5, CD28, OX40, 4-1BB, CD16, Ly49, NKp30 (CD337), NKp44 (CD336), NKp46 (NCR1) or CD3 zeta (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, and the 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 the use of such compositions. For example, such compositions may be compositions comprising 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 be compositions comprising a plurality of different antigen-binding peptides according to the invention, e.g., an antibody 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, e.g., 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. A 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 generalized lipodystrophy. In one embodiment, said anti-perilipin-1 autoantibodies induce complement-mediated destruction of adipocytes in acquired generalized 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. There are different types of lipodystrophies, such as congenital lipodystrophy syndrome and acquired lipodystrophy syndrome. The term "lipoatrophy" typically describes a (near) complete loss of adipose tissue from an area of ​​the body or the entire body. In one embodiment, lipodystrophy is used synonymously with lipoatrophy. In one embodiment, lipodystrophy relates to acquired generalized lipoatrophy (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 family of perilipins, a family of proteins associated with the lipid droplet surface. Perilipin-1 expression is elevated in obese humans. Perilipin-1 protects lipid droplets from lipases involved in lipolysis. In humans, there are three perilipin-1 isoforms, namely perilipin-A, perilipin-B and perilipin-C. The human amino acid sequence of perilipin-1 is according to 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 relates to a portion of an antigen that is recognized by the immune system, in particular an antibody, a B cell or a T cell. 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 the structure and interaction with the paratope. In one embodiment, the antigen perilipin-1 comprises six epitopes, said epitopes having any of the sequences disclosed in SEQ ID NOs: 1-6, 330-333. The antibody or antigen-binding fragment according to 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 according to 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, in particular the perilipin-1 fragment, is presented by MHC molecules on the surface of adipocytes. In one embodiment, said MHC molecule is an MHC-I or MHC-II molecule.

[0086] The term "targeting", as used herein, relates to the ability of a molecular structure, such as an antibody, an antigen-binding peptide, or a surface molecule of a cytotoxic T cell, to bind to an antigen, in particular a certain structure, such as an epitope, by specific interaction. In one embodiment, perilipin-1 fragments at the surface of an adipocyte are targeted by an antigen-binding peptide, which leads to activation of an effector system, which then leads to elimination of said adipocyte, for example, by the caspase cascade. In one embodiment, perilipin-1 fragments at the surface of an adipocyte are targeted by cytotoxic T cells that specifically bind to said perilipin-1, preferably leading to cell death of said adipocyte. In one embodiment, the terms "targeting" and "recognition" are used interchangeably. In one embodiment, the antigen-binding peptide of the present invention targets 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 invention targets perilipin-1 fragments presented by MHC at the cell surface, or such perilipin-1 fragments may be present or presented at the cell surface by another mechanism. In one embodiment, an antigen-binding peptide targets at least one of the perilipin-1 epitopes having any of SEQ ID NOs: 1-6, 330-333. In one embodiment, targeting of perilipin-1 fragments presented in adipocytes using an antigen-binding peptide preferably results in elimination of said adipocytes by an effector / elimination mechanism. In one embodiment, when said antigen-binding peptide binds to two or more of SEQ ID NOs: 1-6 and 330-333, said 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 envisages 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 dual or triple combinations a) to s) described above.

[0089] The term "elimination" as used herein relates to targeting of target cells and induction of cell destruction, for example by the complement cascade or by recruitment of NK cells or by attack from cytotoxic T cells, preferably further including disintegration and / or removal of the resulting cell debris. In one embodiment, the antigen-binding peptide of the invention induces the elimination of adipocytes by effector mechanisms. In one embodiment, targeted adipocytes presenting 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 in a manner sufficient to terminate the cell's ability to survive and / or to terminate the cell's ability to maintain its cellular function. In many of the embodiments, elimination preferably relates to complete removal of adipocytes. In many of the embodiments, "elimination" refers to the destruction and complete degradation of adipocytes, preferably by phagocytosis of said adipocytes, preferably without leaving extracellular cell debris. In one embodiment, perilipin-1 presented on adipocytes is targeted using an antigen-binding peptide to eliminate said adipocytes. In one embodiment, the elimination of adipocytes 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 relate 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 to eradicate cells, such as infected cells. In one embodiment, "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, the 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 said 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) by interaction with Fc receptors expressed by effector cells and / or with 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 said 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 said 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, said recognition being carried out, for example, via the effector cell receptor FcγR and the Fc domain of said antigen-binding peptide, which triggers the effector cells to release cytoplasmic perforin, granulysin and granzymes, thereby inducing apoptosis and / or lysis of said target cells. In one embodiment, the effector mechanism leading to the destruction of adipocytes can also include said cytotoxic drugs, toxins or radionuclides delivered to said adipocytes by the antigen-binding peptides of the present invention, such as by conjugation of said cytotoxic drugs, toxins or radionuclides to the antigen-binding peptides, or by using liposomes labeled with the antigen-binding peptides to deliver said cytotoxic drugs, toxins or radionuclides.

[0092] In one embodiment, the effector mechanism may involve a chimeric antigen receptor (CAR), a receptor protein engineered to give T cells the ability to target specific proteins. In particular, T cells are taken 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 of 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, and then the genetically modified T cells are administered to the patient as CAR-T cells to attack fat cells. In one embodiment, the CAR-T cells can be derived from T cells of 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 altered 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 effector mechanism that results in the destruction and / or elimination of a target cell. In one embodiment, the effector cell may 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 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, said recognition and / or binding induces an effector mechanism that results in the elimination / destruction of said adipocyte. In one embodiment, the effector cell, such as a T cell, recognizes an adipocyte target cell and induces cell death of said target cell. In one embodiment, effector cells such as NK cells, monocytes, macrophages or granulocytes recognize the antigen-binding peptides of the invention bound to adipocytes, and this recognition triggers induction of apoptosis or lysis of said adipocytes and / or phagocytosis of said adipocytes.

[0094] The term "adipocyte" as used herein relates to a fat cell specialized for storing fat. Adipose tissue is mainly composed of adipocytes. There are different types of adipose tissue, namely white fat, brown fat, beige fat and marrow fat, and therefore different types of fat cells. White fat cells are monovacuolar cells that contain one large lipid droplet surrounded by a layer of cytoplasm. Brown fat cells are plurivacuolar and typically polygonal in shape. Beige fat is a combination of white fat cells and Beige fat includes cells of beige and brown fat, making it more metabolically desirable than white fat. There are also adipocytes in bone marrow adipose tissue, which also appear to have 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, the adipocytes preferably present a fragment of perilipin-1 on their surface. In one embodiment, said fragment of perilipin-1 presented on the surface of the adipocyte preferably comprises any 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 by Fc receptor-mediated binding of antigen-binding peptides to perilipin-1 or a fragment thereof results in the destruction of target cells and the removal of their constituents. In one embodiment, the direct elimination mechanism involves neutrophils (neutrophilic polymorphonuclear granular cells) by Fc receptor-mediated binding of antibodies or antigen-binding fragments to perilipin-1 or a fragment thereof. In one embodiment, the direct elimination mechanism preferably comprises macrophages and / or neutrophils binding to antigen-binding peptides attached to perilipin-1 fragments on the surface of adipocytes, said binding resulting in phagocytosis of said adipocytes, said binding preferably being mediated by Fc receptors.

[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 involves another system to destroy said target cell and / or remove the cell remnants. In one embodiment, the indirect elimination mechanism involves the 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, said binding triggers the destruction of said adipocyte by activation of the complement cascade. In one embodiment, the mechanism is complement-dependent cytotoxicity (CDC). In one embodiment, the indirect elimination mechanism involves antibody-dependent cell-mediated cytotoxicity (ADCC), which involves the 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 the 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 involving lysis of adipocytes by effector cells directly binding to perilipin-1 fragments presented via MHC on the surface of said adipocytes. The destruction of adipocytes then triggers the activation of another system, such as phagocytosis, which results in the elimination of the debris of the destroyed adipocytes. In one embodiment, the indirect elimination mechanism involves the phagocytosis of target cell remnants by macrophages or neutrophils (neutrophilic polymorphonuclear granular cells). In one embodiment, the means for specifically targeting and / or eliminating adipocytes can have both direct and indirect elimination capabilities. In one embodiment, macrophages and neutrophils are both direct elimination means, such as by Fc receptor-dependent binding to and phagocytosis of the targeted adipocytes, and indirect elimination means, such as by phagocytosis of the debris of destroyed adipocytes targeted by a different elimination means, such as complement-mediated opsonization. In one embodiment, the direct and indirect elimination mechanisms can occur simultaneously and / or the scope of both can overlap. 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, to T cells, such as cytotoxic T cells or regulatory T cells, to natural killer cells, to B cells, to plasma cells or to 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, said triggering resulting in the elimination of said adipocytes.

[0098] The term "endogenous effector system" as used herein refers to an immune effector system that is endogenous. 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 that are 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 relates 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 donor, or can be obtained from a commercial source. In one embodiment, the exogenous effector system can relate to chimeric antigen receptor T cells (CAR-T cells), which are genetically modified 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 relate to an exogenous T cell receptor (TCR), which can be a soluble TCR or a 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 trigger 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 the membrane attack complex, which forms a transmembrane channel and causes osmotic lysis of the target cell. 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 the cell 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 MHC-I molecules and destroys the cell. The interaction of the T cell with the complex of the antigen and MHC-I molecules of the target cell typically also involves a co-receptor, namely CD8. In one embodiment, upon binding to such a complex on the surface of the target cell, the T cell releases any of the cytotoxins perforin, granzyme and granulysin. In one embodiment, the action of perforin allows granzyme to enter the cytoplasm of the target cell, and its serine protease function triggers the caspase cascade, resulting in apoptosis of said target cell. In one embodiment, apoptosis of said target cell may also be induced by cell surface interactions between cytotoxic T cells and the target cell, such as by Fas-Fas ligand interaction.

[0102] The term "means for specifically targeting perilipin-1 fragment-presenting adipocytes" as used herein relates to any means capable of specifically targeting perilipin-1 fragments presented on the surface of adipocytes and directly and / or indirectly inducing elimination of said adipocytes. In one embodiment, said means for specifically targeting perilipin-1 fragment-presenting adipocytes relates to an antigen-binding peptide capable of specifically recognizing perilipin-1 fragments, preferably via an epitope having any of SEQ ID NOs: 1-6, 330-333. In one embodiment, said means for specifically targeting perilipin-1 fragment-presenting adipocytes comprises and / or induces an effector mechanism that induces elimination of said adipocytes. In one embodiment, said 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, said means for specifically targeting perilipin-1 fragment-presenting adipocytes induces an indirect and / or direct elimination mechanism. In one embodiment, said means for specifically targeting perilipin-1 fragment-presenting adipocytes is preferably a means for specifically eliminating perilipin-1 fragment-presenting adipocytes. In one embodiment, said 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 means 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 a deoxyribonucleic acid. In one embodiment, said nucleic acid encodes an antigen-binding peptide according to other embodiments of the invention. In one embodiment, said nucleic acid encodes a sequence that is at least 50%, preferably at least 85% identical to a nucleic acid sequence that encodes an antigen-binding peptide according to other embodiments of the invention. In one embodiment, said nucleic acid that is at least 50%, preferably at least 85% identical to a nucleic acid sequence that encodes 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 animals", 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 candidates of 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 for the production of the antigen-binding peptide of the invention. In one embodiment, said host cell allows the production of an antibody or an antigen-binding fragment, and includes, for example, a yeast cell or a Chinese Hamster Ovary (CHO) cell. In one embodiment, the host cell is transformed to contain a nucleic acid encoding the antigen-binding peptide of the invention, said transformed host cell being referred to as a recombinant cell. In one embodiment, said 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 has accumulated in the body to an extent that may have a negative effect on the health of the body. In one embodiment, 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, are used to target and / or eliminate adipocytes in obese patients.

[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, the 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, the 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, the binding assay can relate to surface plasmon resonance measurement.

[0111] The term “K D" as used herein refers to the dissociation constant, which is the reciprocal of the association constant. In one embodiment, the antigen-binding peptide of the invention has 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 anticalins or DARPins, 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 has at least 10 -4 Affinity of M (K D 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 point 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 that typically include different antibodies made against different epitopes of an antigen, mAbs are typically made 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 that are free of other immunoglobulin contamination. In one embodiment, the mAb refers to a chimeric, humanized or human antibody or antibody fragment. In one embodiment, the monoclonal antibody can be an optimized monoclonal antibody with an engineered Fc region with improved binding properties to be targeted by effector cells. In one embodiment, said optimized monoclonal antibody can 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 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 antibody that contains sequences derived from non-human immunoglobulin, such as Fab fragments, Fab' fragments, F(ab') 2 The term "humanized antibody" refers to an immunoglobulin chain or fragment thereof, such as a humanized antibody, a IgG1A antibody, a humanized antibody, a humanized antibody, or a humanized antibody fragment thereof, such as ...

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

[0116] The term "substantially intact antibody," as used herein, refers 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 relates to off-target binding to a structure that is not the target structure. In one embodiment, "cross-react" relates 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 between the sequence in question and a polypeptide / nucleic acid sequence of the 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 relates 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 scFv or Fab fragments and contain coding DNA or RNA. The genetic diversity of these libraries is generally 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, the antigen-binding peptides can be optimized to enhance the effector function and / or half-life of said antigen-binding peptides.

[0121] The term "optimized" as used herein relates to enhanced properties of a peptide modified by protein engineering compared to the unmodified form of said peptide. In one embodiment, an antigen-binding peptide is optimized with respect to the strength and / or type of effector function it triggers with an effector system by designing said 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, said Fc region is optimized by genetic engineering to contain amino acid exchanges and / or altered glycosylation compared to an unmodified Fc region. In one embodiment, the serum half-life of an antigen-binding peptide is increased by designing said peptide to include a CH3 region of an IgG domain that increases the half-life or by modifying said antigen-binding peptide with other half-life increasing molecules.

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

[0123] [Figure 1] Labeling of lipid droplet membranes by IgG autoantibodies from a patient with Lawrence syndrome / AGL. With the exception of the fluorescent labeling, visualization was achieved with 10 nm gold particles in electron microscopy (EM) and with 1 nm gold 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 vacuoles, which cannot be distinguished from the cell membrane at this magnification. The loose connective tissue between the fat cells is not stained. 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 the lipid storage droplets is intimately labeled with gold particles. Only isolated gold particles are present in the cell membrane (white arrowheads), whereas no gold particles appear in the nuclear membrane. Bar, 1 μm. E) Labeling is absent 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 plasma membrane of an adipocyte. Nearby are tissue eosinophilic granular cells. C, caveolae; core, lipid core of LSD; EF, outer cytoplasmic cleavage plane; EOSG, eosinophil granule; LSD, lipid storage droplets; N, nucleus; PM, plasma membrane. Bar, 0.5 μm. Images shown are representative of at least three independent experiments. [Diagram 2]Human anti-adipose autoantibodies react with multiple perilipin-1 variants. A) Two-dimensional separation of proteins from rat adipocyte membranes. In the first dimension, proteins were separated in 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 colored 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. Respectively, three out of 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), a result due to differential splicing of the perilipin-1 transcript. Accession numbers are sequence identification numbers in GenBank. The theoretical molecular masses displayed in the fourth column were calculated from the amino acid sequence. The Mascot interface (MASCOT 2.1.03) is used to present PMF spectra for identification and to depict the Mascot score (P=0.05). C) Comparison of a commercial anti-perilipin-1 antibody (Affinity BioReagents, Golden, CO, USA) with human anti-adipose IgG. Immunoblotting was performed after subjecting 10 μg of membrane protein to SDS-PAGE and cutting the PVDF membrane between the two lanes of membrane protein (↓). Incubation of the patient's autoantibodies with anti-perilipin-1 antibody demonstrates an identical pattern of positive bands. [Diagram 3]Autoantibodies do not interfere with lipid storage homeostasis in differentiated human preadipocytes. Human preadipocytes were induced for adipose conversion and allowed to accumulate triacylglycerol over 14 days. Such cells were loaded with fluorescently labeled fatty acids (NBD-FAs) for 3 h in the presence of 10 nM insulin. After washing to eliminate 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-fat IgG (black bars) or control IgG (gray bars), respectively. 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 4 experiments. A, B) Anti-fat 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 among pools of stored triacylglycerol. E,F) Autoantibodies have no effect on the distribution of incorporated NBD-FAs among pools of lipid synthesis and degradation intermediates, diacylglycerol (DAG) and monoacylglycerol (MAG). [Figure 4]Anti-adipose autoantibodies accumulate in 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 normal protocol (L, M). (K) There was no immunoreactivity in fixed cells with control IgG. (A-J,N) Live cells incubated with primary antibodies, (K-M) fixed cells incubated with primary antibodies. (M,N) Overlay of the indicated areas of the fluorescent images (J and L, respectively) and the corresponding phase contrast images; red channel set to 2x intensity for better visibility in the overlaid images. Scale bars: 250 μm (A-L) or 100 μm (M-N). [Diagram 5]Anti-fat 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 until day 8 after induction of differentiation. Cells were preincubated with anti-fat IgG or control IgG for 40 min as indicated. Then, medium was added without (control) or with rabbit complement (final dilution of either 1:64 or 1:32) as seen in the figure. Anti-fat IgG was additionally tested at twice the concentration normally used in the presence of complement 1:32 (anti-fat IgG 2x). A) Cells were fixed and stained with Fluoro Quench 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 versus 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 versus 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 induced complement response on adipocytes mediated by anti-perilipin-1 antibodies. Perilipin-1 peptides are degraded by proteasomes and the resulting perilipin-1 fragments are presented by MHC at the cell surface of adipocytes. Antibodies targeting epitopes having peptide sequences, i.e., any of SEQ ID NOs: 1-6, 330-333 disclosed herein, specifically target adipocytes and allow specific elimination of adipocytes by complement activation (indirect elimination mechanism). Alternatively, cytotoxic T cells specific for at least one perilipin-1 epitope can eliminate adipocytes presenting said perilipin-1 by an indirect elimination mechanism. In addition, other effector systems that can be activated by antibodies binding to perilipin-1 fragment-loaded MHC can use a 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 adipocytes of rat fat pads were separated by SDS-PAGE and stained with Coomassie blue (A) or transferred to PVDF membranes (B). Anti-adipose IgG reacts with two large and several small bands in preparations containing lipid-associated proteins. (C) A control membrane was cut (↓) along the molecular weight standard lane and incubated with either control serum and then 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 for anti-adipose antibodies. 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 proportion of cells are susceptible to autoantibody-induced cell death. Cells were treated in parallel to the experiment summarized in Figure 5, but the incubation lasted 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 a significant loss of cells 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 marked difference due to the detachment of dead cells. (B) Significant differences relative to all respective controls were identified by Student-Newman-Keuls all pairwise multiple comparisons ANOVA (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 arrays of perilipin-1 epitopes. Perilipin-1 epitopes with SEQ ID NOs: 1-6, 330-333 were identified using peptide arrays (PEPperPRINT, PEPperMAP) with perilipin-1 autoantibodies. The sequences of human perilipins 1-5 were extended by 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, and the peptide sequences had an overlap of 14 amino acids each. The peptide arrays thus produced were screened with perilipin-1 autoantibodies as described in the previous examples. Antibody specificity was verified by screening all possible 15mer 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. Thus, 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 of the identified peptides (epitopes) are highly conserved in the mouse and human sequences. [Figure 10-1]Representative flow cytometry fluorescence histograms of anti-perilipin antibodies binding to primary adipocytes (dark grey profile). Antibodies 17-20 (A), 21, 22, 24 and 25 (B) were analyzed. Controls represent cells treated with isotype control and secondary antibody (light grey profile). As a control, a commercial antibody against perilipin-1 from Abcam (ab192716) was used (C). The overlapping histograms observed using the commercial antibody (C) indicate that the commercial anti-perilipin-1 antibody does not specifically bind to adipocytes. In contrast, the antibodies of the 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 of anti-perilipin antibodies binding to primary adipocytes (dark grey profile). Antibodies 17-20 (A), 21, 22, 24 and 25 (B) were analyzed. Controls represent cells treated with isotype control and secondary antibody (light grey profile). As a control, a commercial antibody against perilipin-1 from Abcam (ab192716) was used (C). The overlapping histograms observed using the commercial antibody (C) indicate that the commercial anti-perilipin-1 antibody does not specifically bind to adipocytes. In contrast, the antibodies of the 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 grey profile). Antibodies 26-32 of the invention were analyzed. Controls represent isotype control and cells treated with secondary antibody (light grey profile). Antibodies of the invention bind more efficiently to adipocytes compared to isotope control. [Figure 11-2] Representative flow cytometry fluorescence histograms of anti-perilipin antibodies binding to primary adipocytes (dark grey profile). Antibodies 26-32 of the invention were analyzed. Controls represent isotype control and cells treated with secondary antibody (light grey profile). Antibodies of the invention bind more efficiently to adipocytes compared to 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 addition of a commercial antibody against perilipin-1 from Abcam (ab192716) compared to the control (A) and after 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 to that of the isotope control. The signal obtained for the isotope control is indicated by a dashed line. The commercial antibody tested (PLN1-Ab abcam) is 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 addition of a commercial antibody against perilipin-1 from Abcam (ab192716) compared to the control (A) and after 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 to that of the isotope control. The signal obtained for the isotope control is indicated by a dashed line. The commercial antibody tested (PLN1-Ab abcam) is 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 following the addition of anti-perilipin antibodies such as antibody numbers 20 and 25 (B), compared to a negative control (no antibody added) and an isotype control antibody (A). In the presence of the anti-perilipin antibodies of the invention, particularly antibody numbers 20 or 25, the intensity of live cell staining is decreased and the number of dead cells is increased. Thus, the antibodies of the 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 following the addition of anti-perilipin antibodies such as antibody numbers 20 and 25 (B), compared to a negative control (no antibody added) and an isotype control antibody (A). In the presence of the anti-perilipin antibodies of the invention, particularly antibody numbers 20 or 25, the intensity of live cell staining is decreased and the number of dead cells is increased. Thus, the antibodies of the invention exhibit effective complement-mediated cell death. [Figure 14]Complement-mediated cell death compared to commercial antibodies. Mean values ​​(plus SEM) of the difference in the number of dead cells determined with and without complement are shown; numbers above the bars indicate P values ​​from a two-tailed Student's t-test; with the exception of aF-Ab (a patient-derived polyclonal antibody), all tested commercial antibodies showed non-significant results. The commercial anti-perilipin antibodies tested were PA1-1052 (ThermoFisher), AF6615 (R&D Systems), CST antibody #3470 (D418) and PA5-18693 (ThermoFisher). Thus, commercial antibodies do not demonstrate mediation of 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 different contrast of the total cell overview on the left and the representation of lipid vacuoles on the right indicates that adipocytes are considered. (B) 10x magnification of a 50 μm2 counting section. Left: microvilli, right: AB-bound immunogold particles (white dots). (A+B) For cells incubated with aF-AB. At least 7-8 different cells are included for counting. (C) Quantitative evaluation. ANOVA (P=0.00002) with multiple comparisons (Holm-Sidak, P-values ​​in the figure). An increase in gold particle binding compared to the control is observed for the patient-derived anti-fat antibody (aF-AB) and the antibodies of the invention, such as antibody #5. In contrast to that, 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 than 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 different contrast of the total cell overview on the left and the representation of lipid vacuoles on the right indicates that adipocytes are considered. (B) 10x magnification of a 50 μm2 counting section. Left: microvilli, right: AB-bound immunogold particles (white dots). (A+B) For cells incubated with aF-AB. At least 7-8 different cells are included for counting. (C) Quantitative evaluation. ANOVA (P=0.00002) with multiple comparisons (Holm-Sidak, P-values ​​in the figure). An increase in gold particle binding compared to the control is observed for the patient-derived anti-fat antibody (aF-AB) and the antibodies of the invention, such as antibody #5. In contrast to that, 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 than commercially available anti-perilipin antibodies. [Figure 16-1] Exemplary antigen-binding peptides, particularly antibodies, of the present invention, and their respective CDR sequences. The CDR amino acid sequences of antibodies with antibody numbers 5-11, 13-14, 16-58 are depicted. The CDR sequences of the heavy chain (HCDR) and light chain (LCDR) are shown. The CDR1, 2 and 3 (HCDR) of the heavy chain are depicted from left to right and are each separated by an "_". The CDR1, 2 and 3 (LCDR) of the light chain are depicted from left to right and are each separated by an "_". [Figure 16-2] Exemplary antigen-binding peptides, particularly antibodies, of the present invention, and their respective CDR sequences. The CDR amino acid sequences of antibodies with antibody numbers 5-11, 13-14, 16-58 are depicted. The CDR sequences of the heavy chain (HCDR) and light chain (LCDR) are shown. The CDR1, 2 and 3 (HCDR) of the heavy chain are depicted from left to right and are each separated by an "_". The CDR1, 2 and 3 (LCDR) of the light chain are depicted from left to right and are each separated by an "_". [Figure 17-1]Exemplary antigen-binding peptides, particularly antibodies, of the invention, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies having antibody numbers 5-11, 13-14, 16-58 are depicted. [Figure 17-2] Exemplary antigen-binding peptides, particularly antibodies, of the invention, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies having antibody numbers 5-11, 13-14, 16-58 are depicted. [Figure 17-3] Exemplary antigen-binding peptides, particularly antibodies, of the invention, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies having antibody numbers 5-11, 13-14, 16-58 are depicted. [Figure 17-4] Exemplary antigen-binding peptides, particularly antibodies, of the invention, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies having antibody numbers 5-11, 13-14, 16-58 are depicted. [Figure 17-5] Exemplary antigen-binding peptides, particularly antibodies, of the invention, and their respective VH and VL sequences. The VH and VL amino acid sequences of antibodies having antibody numbers 5-11, 13-14, 16-58 are depicted. EXAMPLES

[0124] Example 1 Isolation of autoantibodies targeting perilipin-1. Written informed consent was obtained from all donors of tissue samples, according to a protocol approved by the Ethics Committee of Jena University Hospital. Stromal vascular population of cells (termed human preadipocytes) from adipose tissue samples were isolated and cultured in medium 199 containing 10% (v / v) fetal bovine serum until confluence [5]. 3T3-L1 cells were obtained from ATCC and maintained in DMEM containing 10% (v / v) fetal bovine serum until confluence. All experiments involving differentiation or stimulation of 3T3-L1 cells were performed in serum-free medium consisting of DMEM and Ham's F12 medium in a 3:1 ratio (without phenol red and with 7.5 mM HEPES, pH 7.2) 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 additionally supplemented with dexamethasone (100 nM) and 3-isobutyl-1-methylxanthine (500 μM). Except where 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-fatty 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. The adipocyte suspension in PBS was shaken thoroughly with half the volume of chloroform. Centrifugation (2500×g) for 5 min at 25° C. separated the lipids dissolved in chloroform from the cytoplasmic proteins in the supernatant and the lipid-associated proteins in the fat cake at the interface of 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 h. Lipid-associated proteins were suspended in PBS by vortexing. Residual chloroform in the final pellet was evaporated in vacuum. 100 μg lipid-associated or cytoplasmic proteins from adipocytes were solubilized with 100 μl sample buffer containing 8 M urea. 10 μl samples were separated in minigels 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 h at 1 mA / cm2 at 8°C. After staining with Ponceau S for 2 min, 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 h: anti-fat IgG and control IgG were used at a dilution of 1:1000, polyclonal rabbit anti-perilipin-1 was used at 1:1250. Protein G-alkaline phosphatase (AP) was diluted 1:5000. Anti-rabbit IgG-AP was diluted 1:1250. Blots were developed with 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 triacylglycerol, 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 times. 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 a patient with lipoatrophy. Serum containing autoantibodies against adipose tissue from AGL patients 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 (Fig. 1A). Besides human unilocular adipocytes, anti-adipose IgG stained adipocytes from all other mammals examined, but not from chicken and carp (data not shown). Immunofluorescence staining of in vitro differentiated 3T3-L1 mouse adipocytes containing multiple lipid storage droplets (LSDs) (Fig. 1B) suggested a specific reaction of the surface of LSDs with anti-adipose IgG. Such specific labeling of the LSD surface was confirmed by transmission electron microscopy in immuno-gold-labeled ultrathin sections of differentiated 3T3-L1 cells (Fig. 1C). The plasma membrane of adipocytes was only sporadically labeled (Fig. 1D), and non-differentiated cells showed no labeling at all (Fig. 1E), suggesting that expression of the autoantigen was 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 a lipid-associated protein identified as multiple perilipin-1 variants. Using isolated adipocytes of rat epididymal fat pads, delipidated and cytosolic fractions of lipid-associated proteins were prepared. In Western blot experiments with anti-fat IgG, a characteristic pattern of two large and several small bands was found almost exclusively in the lipid-associated protein fraction (Figure 7B). The recognition of this band pattern is specific to anti-fat autoantibodies, since neither IgG of healthy controls nor protein A / G-AP alone reacted with these bands (Figure 7C). Thus, the rat adipocyte fraction confirmed the results of the microscopic analysis. After confirming that anti-fat IgG specifically labeled putative autoantigen(s) from adipocytes, the applicants 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-fat IgG (Figure 2A). These spots were isolated and subjected to trypsin digestion, and the peptides were subsequently analyzed by MALDI-TOF-MS. A search in 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 commercial 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 obtain information about the mechanism by which anti-fat IgG causes 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 (Fig. 3A,B). More specific examination of stored triacylglycerols (Fig. 3C,D) and the intermediate products of lipolysis, diacylglycerol and monoacylglycerol (Fig. 3E,F), also did not reveal any effect of anti-fat IgG in in vitro differentiated human adipocytes. Thus, in accordance with the almost exclusive localization of perilipin-1 at the surface of LSDs (Fig. 1), interference of anti-fat IgG with the normal function of perilipin-1 is unlikely to cause adipocyte loss. However, when live 3T3-L1 adipocytes were incubated with the antibodies 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 classical 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 until day 8. Cells were then preincubated in serum-free medium without or with antibodies for 40 min 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 Fluoro Quench AO / EB reagent (OneLambda, Canoga Park, USA) was added. After 15 min, fluorescent images were captured using a FITC filter (long pass emission). Complement dilutions were freshly prepared and activity checked before each individual experiment.

[0131] Example 8 Anti-fat autoantibodies induce complement-dependent cell death in 3T3-L1 cells. We examined whether anti-fat IgG directs the complement-mediated response towards intact adipocytes. Complement alone or in the presence of control IgG had no effect on the staining of dead or viable cells, respectively (Fig. 5). In contrast, within 1 h, anti-fat IgG induced a large increase in the number of dead cells in the presence of complement (Fig. 5A, B). Higher complement concentrations led to a further increase in the number of dead cells, which was reflected by a significant loss of viable cells (Fig. 5C). Extension of the incubation time to 24 h had no effect on cell viability or the number of dead cells in the absence of anti-fat IgG, but induced an even more pronounced loss of viable cells in the presence of anti-fat IgG (Fig. 8). Apparently, the low-density epitopes targeted by anti-perilipin-1 autoantibodies on the plasma membrane of adipocytes (see Fig. 1D) were sufficient to induce complement-mediated cell death. Interestingly, after treatment with complement plus anti-adipose IgG, the remaining viable cell population consisted primarily of preadipocytes lacking LSD and perilipin-1, again indicating a 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 generalized lipodystrophy. Generalized lipodystrophy is associated with severe metabolic complications. Familial forms have been identified as defects in genes key to adipocyte function. The pathogenesis of acquired lipodystrophy is poorly understood, but is often associated with chronic infection or autoimmune markers. As described in previous examples, the applicants isolated autoantibodies against adipose tissue from serum of AGL patients. The immunoglobulin G fraction of patient serum, but not normal serum, specifically stained lipid-accumulating adipocytes in tissue sections and in vitro, pointing to a direct immune reaction against adipocytes involved in the pathogenesis of AGL. Lipodystrophy-associated autoantibodies react with an intracellular autoantigen exclusively expressed in adipocytes and located on the surface of lipid storage droplets, which was identified as perilipin-1 by two-dimensional electrophoresis coupled to 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 has a key role in the selective loss of adipocytes in AGL via a complement-mediated response.

[0133] We disclose that lipodystrophy-associated autoantibodies react with an autoantigen, identified as perilipin-1, that is exclusively expressed in adipocytes and located on the surface of lipid storage droplets. The autoantibodies do not alter cellular lipid homeostasis and do not 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. For antibody selection of perilipin-1 specific antibodies, a naive human antibody-phage library was used. First, non-specific or cross-reactive antibody-phages were removed from the library (campaign a). For that purpose, 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 was synthesized with a biotin-PEG spacer and a GGSGGS linker (SEQ ID NO: 438) at the N-terminus. Additionally, a peptide was synthesized with a biotin-PEG spacer and a AAAAA linker (SEQ ID NO: 439) at the C-terminus. Both peptide variants were refined by HPLC purification.

[0137] First, the target peptide (with 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 non-specific 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, the bacteria were co-infected with M13K07 helper phage to induce antibody-phage amplification. The amplified phages were used for two more 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 with lambda and kappa antibody libraries separately from each other.

[0138] [Table 1]

[0139] To direct the antibody-phage selection towards cell-binding antibodies, the antibody-phage were used for selection on the cell surface after selection cycle 2 of campaign a as well (campaign b). For that purpose, the amplified phages were incubated in the presence of HEK293 cells. The 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. The cells were washed multiple times to remove non-specific or weakly bound antibody-phage particles. The antigen-specific antibody phages were rescued by E. coli infection. An overview of the antibody selection strategy is given below. Each strategy was performed separately from each other 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 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] In campaign b, 18 antibodies were identified that were identified in parallel in campaign a:

[0146] [Table 4]

[0147] Based on the 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. VH genes were inserted into mammalian human IgG1 heavy chain expression vector. VL genes were inserted into mammalian human lgG1 light chain lambda or kappa expression vector. IgG expression vectors were transformed into E. coli. Single colonies were selected for plasmid expansion. After isolation of transfection grade DNA, HEK cells were transiently co-transfected with 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 adjustment of the buffer to PBS, the protein concentration of the antibodies was determined by UV / VIS spectrometry. The integrity and purity of the antibodies 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 tested for their functional characteristics. In particular, the binding and functional characteristics of antibodies with various VH and VL regions were tested (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, in particular 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), in particular any of SEQ ID NOs: 330, 331, 332 and 333, respectively.

[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. Adipocytes were freshly isolated from WT untreated mice and stained with selected antibodies. In particular, 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 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. The adipocytes were then 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 non-specific binding of the antibody. Moreover, one control sample was incubated only with the secondary antibody without the addition of the primary antibody. After washing, the adipocytes were incubated with a secondary antibody (goat anti-human Alexa 568) that recognizes the primary antibody. After incubation and an additional washing step, the DNA stain Hoechst 33342 was added at a 1:1000 dilution to identify intact cells in flow cytometry. Flow cytometry was then used to analyze the binding of the anti-perilipin antibody to the adipocytes.

[0154] For signal detection, anti-human Alexa 568 was used, which recognizes the primary antibody (Figures 10 and 11). Non-specific 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 antibodies compared to the isotype control. Histograms of the tested antibodies of the invention showed enhanced binding of the antibodies of the invention to adipocytes compared to the binding of the isotype control to adipocytes. Furthermore, the antibodies of the invention showed enhanced binding to adipocytes compared to the commercially available anti-perilipin antibody (Figures 10 and 11).

[0155] Various antibodies of the invention were tested for their binding efficiency on the cell surface of adipocytes. The binding of the antibodies of the invention was further analyzed using gold nanoparticles and electron microscopy essentially following the 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 min. The fixed cell layer was then incubated overnight at 4°C without primary antibodies or with 1 μg of each tested antibody per coverslip. After washing, the coverslips were incubated for 5 h at 4°C with a crosslinking antibody: rabbit anti-human IgG (DAKO; Cat. No. A0423; for any human or humanized primary antibody) or polyclonal mouse anti-rabbit (H+L) (Invitrogen; Cat. 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-20 nm; catalog number EMGAR20; for human or humanized primary antibodies; or goat anti-mouse Au-20 nm; catalog number EMGAM20; for commercial primary antibodies). After a final wash, intact adipocytes were identified by their lipid vacuoles and microvilli, and bound gold particles were detected in field emission scanning electron microscopy mode.

[0156] Successful binding of the antibodies of the invention to adipocytes was observed. Binding of the antibodies of the invention, for example, antibody 5, to adipocytes is shown in Figure 15. Similar results were observed for other antibodies of the invention, for example, antibodies 11, 17, 20, 29 and 31 (data not shown). In contrast, electron microscopy did not show 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 was prepared by mixing DMEM (NaHCO 3 DMEM / HAM's F12 medium (1:1) containing 15 mM HEPES mixed 1:1 with 1 g / l glucose (without 1 g / l glucose), 1.2 g / l NaHCO 3 , 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 for 4 days in SFD medium supplemented only with insulin (1 μM). For complement assays

[10] , cells were incubated with selected antibodies (anti-perilipin and isotype control antibodies) at a final concentration between 0.017 and 0.05 mg / ml in 60 μl SFD medium (pooled antibodies each at a concentration of 0.0017 mg / ml per antibody in 60 μl 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 on top of the cells. After 1 h 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 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 available from Thermo Fisher Scientific Inc.

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

[0160] (Example 16) Further evaluation of exemplary antibodies of the invention. Various antibodies of the invention were further investigated with respect to 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 explore 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 may, both separately and in any combination thereof, 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) An antigen-binding peptide according to any of the preceding items, which is not a perilipin-1 targeting autoantibody of a human patient with lipoatrophy, preferably not a perilipin-1 targeting anti-PLIN1 antibody against a peptide spanning Thr8 to Ala145 having manufacturer R&D catalogue #AF6615. (Item 5) 13. 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 ) an antigen-binding peptide according to any of the preceding items that binds to perilipin-1 at (Item 7) 13. The antigen-binding peptide of any of the preceding items, which is an antibody, preferably a monoclonal antibody. (Item 8) An antigen-binding peptide according to any of the preceding items, wherein the antigen-binding peptide, in particular 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. (Item 9) 13. The antigen-binding peptide of any of the preceding items, wherein when the antigen-binding peptide, in particular the antibody, is incubated with adipocytes, the antigen-binding peptide, in particular 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. comprises 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. comprises 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. comprises 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. comprising 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. comprising 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 a peptide comprising an amino acid sequence whose heavy chain CDR1, CDR2, CDR3 regions and whose light chain CDR1, CDR2, CDR3 regions have at least 90% identity, preferably at least 95% identity, more preferably at least 98% identity, even more preferably at least 99% identity to any of the sequences set out in i. to lii. or an antibody. (Item 11) 13. 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 antibodies, substantially intact antibodies, Fab fragments, F(ab') 2 2. 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 claims, capable of binding to perilipin-1 fragments presented on adipocytes, wherein the binding of the antigen-binding peptide to perilipin-1 fragments on the surface of the adipocytes activates the immune effector system to target and / or eliminate the adipocytes. (Item 14) 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 an effector cell 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) 13. An antigen-binding peptide according to 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 one of SEQ ID NOs: 1 to 6; (ii) a second binding site; and 2. A bispecific antigen-binding peptide comprising: (Item 18) 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 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. (Item 19) 19. The bispecific antigen-binding peptide according to 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 of items 1 to 15 and / or the bispecific antigen-binding peptide according to any of items 17 to 18. (Item 21) The composition according to item 20, comprising two or more antigen-binding peptides according to any of items 1 to 15 and / or two or more bispecific antigen-binding peptides according to any 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) The composition comprises at least two or at least three or at least 4 or at least 5 or at least 6 antigen-binding peptides, each of which binds to a different perilipin-1 epitope, and / or the composition comprises at least 2 or at least 3 or at least 4 or at least 5 or at least 6 bispecific antigen-binding peptides according to any of items 17-18, 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 of items 1 to 15 and / or the bispecific antigen-binding peptide according to any of items 17 to 18 for use in the prevention or treatment of obesity. (Item 26) 19. An isolated nucleic acid encoding the antigen-binding peptide according to any one of items 1 to 15, or encoding the bispecific antigen-binding peptide according to item 17 or 18. (Item 27) 27. A recombinant cell comprising the nucleic acid according to 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: 1-6, 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 harvesting cells expressing antigen-binding peptides from the spleen of said non-human animal, preparing hybridoma cells therefrom, expressing said antigen-binding peptides, 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: 1-6 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 pool of cells containing cytotoxic T cells, such as by tetramer staining or dextramer 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 a 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; The method includes: (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

[Claim 1] The invention described in the specification.

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