Linear 23 peptide capable of coupling single B cell epitope or hapten to prepare immunomodulating peptide and antibody and application thereof

An immunoregulatory, B cell technology, applied in the fields of immunology, pharmacy and medicine, to improve immunogenicity, improve symptoms, and increase CD4+

Inactive Publication Date: 2018-10-02
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How to select the appropriate T helper cell (Th) epitope and how to link with B cell epitope or peptide hapten to significantly improve the immunogenicity of B cell epitope or peptide hapten is a world problem

Method used

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  • Linear 23 peptide capable of coupling single B cell epitope or hapten to prepare immunomodulating peptide and antibody and application thereof
  • Linear 23 peptide capable of coupling single B cell epitope or hapten to prepare immunomodulating peptide and antibody and application thereof
  • Linear 23 peptide capable of coupling single B cell epitope or hapten to prepare immunomodulating peptide and antibody and application thereof

Examples

Experimental program
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Effect test

Example Embodiment

[0027] Example 1: Preparation of 23-peptide LP1 containing T helper epitopes (Th epitopes).

[0028] The 23 peptide was designated LP1, and the amino acid sequence is shown in SEQ ID No. 1. LP1 is transformed from a Th2 cell epitope. This Th2 cell epitope is derived from a P277 peptide at positions 437 to 460 on the human heat shock protein HSP60. The N-terminal of the P277 peptide has a Th2 composed of 13 amino acids. Epitope P2, a linking peptide consisting of 10 amino acid residues is added to its N-terminus to form LP1. The amino acid sequence of the linking peptide consisting of 10 amino acid residues is shown in SEQ ID No. 2, named L1, which contains A B cell epitope derived from human islet cell-specific antigen-insulinoma-associated protein-2 (IA-2) near membrane region JM2, its amino acid sequence is: FEYQD, named IA2(5), at its N-terminus Add amino acid Gly, add 4 amino acid sequence peptide SGTI at the C-terminus to form L1.

Example Embodiment

[0029] Example 2: Preparation of B cell epitopes or synthetic peptide haptens.

[0030] Using a variety of B cell antigen epitope analysis software and online analysis tools, based on a single parameter, combined with secondary structure prediction and spatial structure characteristics, respectively, from a target antigen protein (such as: human DPP4 or human XOD or human uric acid transporter protein). 1) Select a number of dominant B cell antigen epitopes or peptide haptens from more than 100 epitopes. By connecting with 23 peptides, through the process of screening, re-engineering and re-screening in immunized animal experiments, a highly immunogenic B cell can be obtained. B cell epitopes or synthetic peptide haptens.

Example Embodiment

[0031] Example 2-1: Preparation of the dominant B cell antigen epitope of human dipeptidyl peptidase 4 (dipeptidyl peptidase 4, DPP4) by selecting multiple predicted dominant B cell antigen epitopes from its more than 100 epitopes B-cell epitopes or peptide haptens with high immunogenicity such as D1 are obtained through screening of immunized animal experiments. D41 is a polypeptide sequence 88VFLENSTFDE97 of DPP4, with a total of 10 amino acid residues, and the amino acid sequence is shown in SEQ ID No.3.

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Abstract

The invention provides a linear 23 peptide molecule. The linear 23 peptide molecule can be used for coupling a single B cell epitope or hapten to prepare immunomodulating peptide. The immunomodulatingpeptide immunizes animals and can produce an antibody with a high titer that can specifically bind to a protein containing a B cell epitope. A Th2 antibody or a single-chain antibody can be preparedby a hybridoma technology, or an antibody library or single-cell and binding genetic engineering and other technologies. The immunomodulating peptide and the antibody both significantly enhance the immunoregulation deflecting toward Th2, inhibit the activity or function of protease containing the B cell epitope, and improve related diseases. At the same time, the 23 peptide and a coupling or fusion protein or a composition/polymer thereof provided by the invention can also be used in recombinant microorganisms and transgenic animals or plants and cells thereof, so that the 23 peptide and the coupling or fusion protein or the composition/polymer thereof can be produced in production plants to produce vaccines or oral vaccines or produce specific antibodies, and the above peptides and antibodies can be used in the development of drugs to prevent and treat diseases with high Th1, including but not limited to diabetes, hyperuricemia and complications, as well as in the development of detection reagents to detect proteins or peptides containing B cell epitopes.

Description

technical field [0001] The invention relates to the related fields of immunology, pharmacy and medicine. The present invention relates to a linear 23-peptide capable of coupling a single B-cell epitope or hapten for preparing immunomodulatory peptides and antibodies and its use. It is transformed from a Th2 cell epitope, and 23-peptide molecules are covalently linked A single B-cell epitope or hapten, such as a B-cell epitope of DPP4, a B-cell epitope of xanthine oxidase, constitutes an immunomodulatory peptide, and the immunomodulatory peptide immunizes animals, which can produce high titers of energy and containing this Protein-specific binding antibodies to B cell epitopes, and animals immunized with immunomodulatory peptides can prepare various antibodies such as Th2 monoclonal antibodies and single-chain antibodies through hybridoma technology, antibody library, single cell and genetic engineering. Both immunomodulatory peptides and antibodies can significantly enhance T...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N9/02C12N9/48C07K16/40A61K39/00A61K39/395A61P3/10A61P3/06A61P13/12A61P9/10A61P19/06A61P19/02A61P29/00A61P25/28A61P27/02A61P9/00
CPCA61K39/0005A61K2039/505A61P3/06A61P3/10A61P9/00A61P9/10A61P13/12A61P19/02A61P19/06A61P25/28A61P27/02A61P29/00C07K14/47C07K16/40C07K2317/76C07K2319/40C12N9/0093C12N9/485C12Y117/03002C12Y304/14005
Inventor 李泰明顾小骞焦瑞
Owner CHINA PHARM UNIV
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