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Screening of peptide mimetic peptides against human histamine receptor 4 (hr4) epitope using phage antibody library and its vaccine construction method

A technology of epitope-mimicking peptides and construction methods, which can be applied to antibody medical components, peptides, drug combinations, etc., and can solve problems such as expensive prices and unclear effects on liver and kidney

Active Publication Date: 2018-08-03
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

JNJ-7777120 is chemically synthesized, expensive, 3030RMB / 50mg, has oral bioavailability, and the effect on liver and kidney is not yet clear

Method used

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  • Screening of peptide mimetic peptides against human histamine receptor 4 (hr4) epitope using phage antibody library and its vaccine construction method
  • Screening of peptide mimetic peptides against human histamine receptor 4 (hr4) epitope using phage antibody library and its vaccine construction method
  • Screening of peptide mimetic peptides against human histamine receptor 4 (hr4) epitope using phage antibody library and its vaccine construction method

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Embodiment Construction

[0035] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the present invention Examples, not all examples. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0036] 1. Main experimental materials

[0037] (1) Target molecule: Anti-HR4antibody (ab178704) monoclonal antibody was purified by Protein A / G in the early stage to obtain a high-purity (>95%) monoclonal antibody that can be used for phage peptide library screening.

[0038] (2) Phage peptide library: phage random dodecapeptide library (Ph.D-12TM phage display peptide library) ...

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Abstract

The invention discloses a method for screening human histamine receptor 4 (HR4) epitope mimetic peptides using a phage antibody library and a vaccine construction method thereof. A polypeptide fragment is obtained by screening a phage random dodecapeptide library, and its amino acid sequence is: FNKWMDCLSVTH. The present invention utilizes the phage random dodecapeptide library technology to screen the polypeptide sequence of the HR4 epitope mimetic peptide, ELISA to identify the affinity of the phage monoclonal to the HR4 monoclonal antibody, obtain a polypeptide sequence named PT1 by sequencing, and further identify the target of the phage positive clone. The tropism results suggest that the phage-positive clones can specifically bind to human HR4 monoclonal antibodies, and the obtained human HR4 monoclonal antibody-specific polypeptides are screened to construct HR4 epitope mimic peptide vaccines. The HR4 mimic epitope-based vaccine of the present invention will It will lay a good foundation in the clinical treatment of allergic rhinitis and the development of HR4 antagonists, and provide a preliminary experimental basis for its construction.

Description

technical field [0001] The invention relates to the technical field of medicine and biology, in particular to a method for screening human histamine receptor 4 (HR4) epitope mimic peptides using a phage antibody library and a vaccine construction method thereof. Background technique [0002] Research on HR4 molecules and their antagonists: 1. HR4 molecule histamine receptor-4 (histamine receptor-4, HR4) is a newly discovered histamine receptor, which is a transmembrane G protein-coupled receptor (G -protein-coupled receptors, GPCRs), the function is to inhibit adenylyl cyclase activity, activate phospholipase C (phospholipase C) and induce calcium ion flow. HR4 is preferentially distributed on immune organs and hematopoietic cells, and is involved in all immune cells in the pathological immune response of allergic rhinitis (allergicrhinitis, AR). The expression of HR4 can be detected in mast cells, CD4+T, CD8+T cells, eosinophils, NK cells, dendritic cells, and monocytes, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08G01N33/577A61K39/00A61P37/08A61P11/02
Inventor 李琳李东原
Owner JILIN UNIV