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Antibody of G protein coupled receptor, preparation method of antibody and G protein coupled receptor kit

A technology for the preparation of coupled receptors and antibodies, used in immunoglobulins, chemical instruments and methods, anti-animal/human immunoglobulins, etc.

Active Publication Date: 2020-05-15
苏州睿瀛生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing methods for detecting GPCRs are generally antibody production techniques using polypeptides as immunogens. For the detection of changes in the content of a specific GPCR, the detection results have limitations, and it is often necessary to purchase a large number of antibodies or kits to detect changes in GPCRs. downstream experiments

Method used

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  • Antibody of G protein coupled receptor, preparation method of antibody and G protein coupled receptor kit
  • Antibody of G protein coupled receptor, preparation method of antibody and G protein coupled receptor kit
  • Antibody of G protein coupled receptor, preparation method of antibody and G protein coupled receptor kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of Antibody I for G Protein-Coupled Receptor

[0048] Step 1. Select the following polypeptide immunogens as the antigen polypeptide group of two or more G protein-coupled receptors with similar homology, and modify the N-terminal and C-terminal of each polypeptide immunogen with cysteine ​​and arginine respectively , purified to obtain the antigenic determinant polypeptide;

[0049] The amino acid sequence of the polypeptide immunogen is:

[0050] FZD8 / FZD5 CNHDTQDEAGLEVHQFWR;

[0051] GPR109 / HCAR2-CLQRKMTGEPDNNRSTR;

[0052] OR10C1 / OR10C1 CHLLTGRRHISRSR;

[0053] OR2A5 / 2A14CKSRHPEEQQKVLR;

[0054] OR2AG1 / 2 CADFLRRENTISFR;

[0055] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0056] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0057] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0058] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0059] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0060] OR2A4 / 7CVGPRYGNPKEQKR;

[0061] OR2AG1 / 2 CLADFLRRENTISFR;

[0062] OR2T2 / 35CQDLLSKDKTISFR;

[0063]OR2T3 / 34 CLRSMMQSRM...

Embodiment 2

[0070] Example 2 Preparation of Antibody II for a G Protein-Coupled Receptor

[0071] The preparation method is the same as in Example 1, except that the following polypeptide immunogen is selected as the antigen polypeptide group of two or more G protein-coupled receptors with similar homology in step 1. The amino acid sequence of the polypeptide immunogen is:

[0072] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0073] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0074] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0075] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0076] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0077] OR2A4 / 7CVGPRYGNPKEQKR;

[0078] OR2AG1 / 2 CLADFLRRENTISFR;

[0079] OR2T2 / 35CQDLLSKDKTISFR;

[0080] OR2T3 / 34 CLRSMMQSRMNQEKR;

[0081] OR4A4 / 47 CYTLRNSEMTSAMKKLR;

[0082] OR8B2 / B3 CHIKSTQGRSKAFR;

[0083] OR11H1 / 11H2 / 11H12 CSLQNKEIKAALRKR;

[0084] OR13C2 / 13C9 CMKPKSKETLNSDR;

[0085] OR2T5 / 2T29 VNKVSAPECGMQMR;

[0086] OR10H3 / 10H4 CYLKPKGLHSMYSDR;

[0087] OR10K1 / 10K2 CDLLSQKKTISFLR;

[0088] OR1D4 / 5 CQMPSASK...

Embodiment 3

[0094] Example 3 Preparation of Antibody III for a G Protein-Coupled Receptor

[0095] The preparation method is the same as in Example 1, except that the following polypeptide immunogen is selected as the antigen polypeptide group of two or more G protein-coupled receptors with similar homology in step 1. The amino acid sequence of the polypeptide immunogen is:

[0096] OR2A5 / 2A14CKSRHPEEQQKVLR;

[0097] OR2AG1 / 2 CADFLRRENTISFR;

[0098] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0099] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0100] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0101] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0102] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0103] OR2A4 / 7CVGPRYGNPKEQKR;

[0104] OR2AG1 / 2 CLADFLRRENTISFR;

[0105] OR2T2 / 35CQDLLSKDKTISFR;

[0106] OR2T3 / 34 CLRSMMQSRMNQEKR;

[0107] OR4A4 / 47 CYTLRNSEMTSAMKKLR;

[0108] OR8B2 / B3 CHIKSTQGRSKAFR;

[0109] OR11H1 / 11H2 / 11H12 CSLQNKEIKAALRKR;

[0110] OR13C2 / 13C9 CMKPKSKETLNSDR;

[0111] OR2T5 / 2T29 VNKVSAPECGMQMR;

[0112] OR10H3 / 10H4 CYLKPKGLH...

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Abstract

The invention relates to the technical field of biological detection reagents, in particular to an antibody of a G protein coupled receptor, a preparation method of the antibody and a G protein coupled receptor kit. Antigenic determinant polypeptide is coupled to hemocyanin to form a holoantigen, and then a polyclonal antibody technology is adopted to prepare the antibody of the G protein coupledreceptor; and the antigenic determinant polypeptide comprises multiple polypeptide immunogens, the N ends of the polypeptide immunogens are modified with cysteine, and the C ends of the polypeptide immunogens are modified with arginine. The antibody of the G protein coupled receptor can recognize G protein coupled receptors with similar homology at the same time, and the G protein coupled receptorkit produced by using the antibody is used for rapidly detecting a cell-level G protein coupled receptor signal.

Description

technical field [0001] The invention relates to the technical field of biological detection reagents, in particular to an antibody to a G protein-coupled receptor, a preparation method thereof, and a G protein-coupled receptor kit. Background technique [0002] G protein-coupled receptors (GPCRs) are the most known family of membrane receptors, play important biological roles in physiological and pathological processes, and are the main superfamily of drug targets. The main function is to mediate extracellular signals into the cell, and participate in autocrine, paracrine, endocrine system signals a series of important reactions inside the cell. About 50% of small molecule therapeutic targets are GPCRs, and the ligands of GPCRs include a wide range, including proteins, peptides, carbohydrates, lipids and so on. For most GPCRs, the external ligand binds to the receptor in the membrane, causing a structural change that activates several or hundreds of guanylate trimers in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C07K7/08G01N33/74
CPCC07K16/2869C07K7/08G01N33/74G01N2333/726
Inventor 邓金亮
Owner 苏州睿瀛生物技术有限公司
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