Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for identifying MHC I binding peptide motif

An identification method and a technology for binding peptides, applied in the field of biomedicine, can solve the problems of long time and low accuracy of peptide binding motif determination, and achieve the effects of efficient screening, short time-consuming and reliable results.

Active Publication Date: 2019-04-23
CHINA AGRI UNIV +1
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problems of long time and low accuracy in the determination of polypeptide binding motifs, the present invention provides a method for identifying MHC I binding polypeptide motifs based on a random peptide library, the biggest advantage of which is short detection time and low cost

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying MHC I binding peptide motif
  • Method for identifying MHC I binding peptide motif
  • Method for identifying MHC I binding peptide motif

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045]Example 1 Porcine SLA I polypeptide motif identification and synthesis of binding polypeptide

[0046] 1.1 Synthesis of random 9-peptide

[0047] Using the Fmoc method to synthesize a 9-peptide library with random sequences, the specific steps are as follows:

[0048] (1) Polypeptide-solid phase carrier cross-linking: 19 kinds of α-amino protecting group amino acids (except cysteine) were prepared by Fmoc (9-fluorenylmethoxycarbonyl), in DMSO at 20-25°C Reaction with alkoxybenzyl alcohol resin;

[0049] (2) Combined deprotection: 19 kinds of cross-linked polypeptide-solid phase carriers were thoroughly mixed, and the amino groups were deprotected by TFA (trifluoroacetic acid);

[0050] (3) Neutralization and washing: neutralize the free amino terminal with triethylamine and fully wash;

[0051] (4) Grouping and condensation: Divide the washed mixture into 19 equal parts, activate the carboxyl group of a new round of amino acids through DCC, and add 19 kinds of amino a...

Embodiment 2

[0105] Example 2 Duck Anpl - MHC I peptide motif identification

[0106] According to the method of Example 1, random sampling was performed on ducks from a duck breeding farm, and lymphocytes were separated by lymphocyte separation medium (Tianjin Haoyang Biological Products Technology Co., Ltd.), and the separation method was carried out with reference to the instructions. The obtained cDNA was reverse-transcribed, and the primer sequences are shown in Table 2:

[0107] Table 2 Anpl -MHC I α chain and β chain primers

[0108] .

[0109] According to the method of Example 1, using cDNA as a template to obtain two Anpl -MHC I, and eventually two Anpl - MHC I binding peptides, and perform LC-MS / MS mass spectrometry sequencing and analysis and data analysis. The result is as Figure 7-13 As shown, the analysis obtained two Anpl - MHC I preference for binding to different sites of the polypeptide. Depend on Figure 13 and Figure 14 It can be seen that Anpl -...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for identifying an MHC I binding peptide motif. The method includes the following steps that (1) a random peptide with the random sequence and the length of 8-12 is synthesized; (2) amino acid distribution is obtained through DeNovo analysis of the random peptide; (3) an alpha chain and a beta<2m> chain of MHC I are expressed; (4) the random peptide, the alpha chainand the beta<2m> chain form a MHC I-peptide compound in a solution; (5) the MHC I-peptide compound is subjected to thermal denaturation, and an MHC I binding peptide is obtained after separation andpurification; and (6) amino acid distribution of the MHC I binding peptide is obtained through DeNovo analysis of the MHC I binding peptide, and by comparing with amino acid distribution in the step (2), the preference of amino acids at different sites of the MHC I binding peptide and main anchor positioning are obtained. According to the method for identifying the MHC I binding peptide motif, operation is simple, the result is reliable, and the consumed time is short, efficient screening of potential pathogen specific T cell epitopes is achieved, and the method can be widely used in preliminary screening of animal or human body peptide vaccines.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for identifying MHCI binding polypeptide motifs by using a random peptide library. Background technique [0002] Major histocompatibility complex class I (Major histocompatibility complex class I, MHCI) molecule-mediated cytotoxic T lymphocyte (Cytotoxic T lymphocyte, CTL) response is an important means for the body to clear diseased cells in the body and resist virus infection. MHC I molecules are expressed on the surface of all nucleated cells and platelets, and bind and present intracellular antigen polypeptides (such as viral polypeptides degraded by proteasomes) for CD8 + The T cell receptor (TCR) on the surface of T cells specifically recognizes and activates CD8 under the cooperation of co-receptor molecules (CD8 molecules, etc.) + T cells specifically kill target cells. This process is called CTL response, and the polypeptides presented by MHC I molecule...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C07K7/06C07K1/06C07K1/04
CPCC07K7/06G01N33/6818G01N33/6848
Inventor 张念之张琳吴家强魏孝辉李卓琳
Owner CHINA AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com