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BP180 single chain antibody of humanized anti-bullous pemphigoid antigen and its use

A BP180, pemphigoid technology, applied in application, antibody, anti-animal/human immunoglobulin, etc., to achieve the effect of treating bullous pemphigoid

Inactive Publication Date: 2011-08-10
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0004] The object of the present invention is to provide a human anti-bullous pemphigoid antigen BP180 single chain antibody (human anti-BP180 scFv), which can be soluble expressed in E. Bullous pemphigoid antigen BP180 single chain antibody solves the problem of clinical treatment of bullous pemphigoid, and eventually becomes a biopharmaceutical for the treatment of bullous pemphigoid

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Embodiment Construction

[0017] The pathogenesis of bullous pemphigoid (BP) is a complex process of severe immune damage to the skin mediated by pathogenic autoantibodies and complement and other inflammatory components. BP180 is epidermal basal cell transmembrane collagen, also known as type XVII collagen (COL17) or bullous pemphigoid antigen 2 (BPAG2), and its gene is located at 10q24.3. The BP180 molecule consists of three parts: the intracellular domain, the transmembrane region and the extracellular domain. The extracellular domain includes 15 collagen coding regions (separated by 15 non-collagen coding regions) and the sixteenth non-collagen coding region. The coding region (NC16A), forms a rod-shaped collagen structure with a flexible tail that connects the basal cells and extends down to the lamina densica before its ends fold back up to the zona pellucida. With this unique structure, BP180 molecule, as an important component of the "hemidesmosome-anchor fiber complex", plays a vital role in m...

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Abstract

The invention discloses a human-derived anti-bullous pemphigoid antigen BP180 ScFv; the gene sequence of the BP180 ScFv structurally belongs to the single-chain antibody (scFv) and is composed of a light chain variable region gene and a heavy chain variable region gene. The antibody preparation method mainly comprises the following steps: a, separating the peripheral blood mononuclear cells of bullous pemphigoid patients, extracting the RNA and constructing a phage antibody library through RT-PCR amplification of the antibody light and heavy chain variable region genes; b, selecting the BP180phage antibodies from the phage antibody library; c, obtaining the human-derived anti-BP180 ScFv of soluble expression through genetic engineering means; d, identifying the binding activity, specificity, affinity and other immunological characteristics of the obtained antibody. The antibody of the invention can block the binding activity between the bullous pemphigoid autoantibody and the corresponding self-antigen and can block the activation of the complement-mediated inflammatory response as a result of the combination of the autoantibodies, and can be used in bullous pemphigoid treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a human-derived anti-bullous pemphigoid antigen BP180 single-chain antibody, which blocks bullous pemphigoid autoantibodies and corresponding autoantigens by combining with BP180 The combination of BP180 can block the inflammatory response mediated by the activation of complement due to the combination of autoantibodies, and has the effect of treating bullous pemphigoid. Background technique [0002] Bullous pemphigoid (BP) is a common and stubborn autoimmune blistering disease, and it is a difficult disease. At present, there is no specific treatment method and the systemic immunosuppression mainly depends on glucocorticoids. [0003] The main pathological change of bullous pemphigoid is the combination of pathogenic autoantibodies, mainly anti-BP180 autoantibodies, with corresponding antigens, activating complement and triggering immune inflammatory response, leading to the separat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N15/13A61K39/395A61P37/06
Inventor 王刚孙娜娜刘玉峰
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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