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Antibody to g protein-coupled receptor, preparation method thereof, and g protein-coupled receptor kit

A technique for coupling receptors and antibody preparation, applied in immunoglobulins, chemical instruments and methods, anti-animal/human immunoglobulins, etc.

Active Publication Date: 2021-12-14
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 to g protein-coupled receptor, preparation method thereof, and g protein-coupled receptor kit
  • Antibody to g protein-coupled receptor, preparation method thereof, and g protein-coupled receptor kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Preparation of Antibody I for a G Protein-Coupled Receptor

[0047] Step 1. Select the following polypeptide immunogens as the antigen polypeptide group of two or more G protein-coupled receptors with similar homology, modify each polypeptide immunogen with cysteine ​​and arginine, purify, and obtain antigenic determination cluster polypeptide;

[0048] The amino acid sequence of the antigenic determinant polypeptide is:

[0049] FZD8 / FZD5 CNHDTQDEAGLEVHQFWR;

[0050] GPR109 / HCAR2-CLQRKMTGEPDNNRSTR;

[0051] OR10C1 / OR10C1 CHLLTGRRHISRSR;

[0052] OR2A5 / 2A14CKSRHPEEQQKVLR;

[0053] OR2AG1 / 2 CADFLRRENTISFR;

[0054] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0055] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0056] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0057] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0058] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0059] OR2A4 / 7CVGPRYGNPKEQKR;

[0060] OR2AG1 / 2 CLADFLRRENTISFR;

[0061] OR2T2 / 35CQDLLSKDKTISFR;

[0062] OR2T3 / 34 CLRSMMQSRMNQEKR.

[0063] Step 2, coupl...

Embodiment 2

[0069] Example 2 Preparation of Antibody II for a G Protein-Coupled Receptor

[0070] The preparation method is the same as in Example 1, except that the amino acid sequence of the epitope polypeptide prepared in step 1 is:

[0071] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0072] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0073] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0074] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0075] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0076] OR2A4 / 7CVGPRYGNPKEQKR;

[0077] OR2AG1 / 2 CLADFLRRENTISFR;

[0078] OR2T2 / 35CQDLLSKDKTISFR;

[0079] OR2T3 / 34 CLRSMMQSRMNQEKR;

[0080] OR4A4 / 47 CYTLRNSEMTSAMKKLR;

[0081] OR8B2 / B3 CHIKSTQGRSKAFR;

[0082] OR11H1 / 11H2 / 11H12 CSLQNKEIKAALRKR;

[0083] OR13C2 / 13C9 CMKPKSKETLNSDR;

[0084] OR2T5 / 2T29 VNKVSAPECGMQMR;

[0085] OR10H3 / 10H4 CYLKPKGLHSMYSDR;

[0086] OR10K1 / 10K2 CDLLSQKKTISFLR;

[0087] OR1D4 / 5 CQMPSASKKYKTFR;

[0088] OR1D4 / 5 CLQMPSASKKYKTFSR;

[0089] OR3A2 / 3 CRLLSHKSTISYDR;

[0090] OR5M1 / 5M10 CMYVRPPSEKSVER;

[0091] OR10X1 / OR10R3P ...

Embodiment 3

[0093] Example 3 Preparation of a G protein-coupled receptor antibody III

[0094] The preparation method is the same as in Example 1, except that the amino acid sequence of the epitope polypeptide prepared in step 1 is:

[0095] OR2A5 / 2A14CKSRHPEEQQKVLR;

[0096] OR2AG1 / 2 CADFLRRENTISFR;

[0097] OR2T2 / 2T35 CHRMNSAEGRRKAFR;

[0098] OR2T2 / 2T35 CLQDLLSKDKTISFR;

[0099] OR4F4 / 4F5 / 4F17 CDFFSQRKVISFKR;

[0100] OR5AU1 / OR5AU1 CLKMSSAQGRFKAFR;

[0101] OR1S1 / 1S2 CPSSTHPEDTDKIGR;

[0102] OR2A4 / 7CVGPRYGNPKEQKR;

[0103] OR2AG1 / 2 CLADFLRRENTISFR;

[0104] OR2T2 / 35CQDLLSKDKTISFR;

[0105] OR2T3 / 34 CLRSMMQSRMNQEKR;

[0106] OR4A4 / 47 CYTLRNSEMTSAMKKLR;

[0107] OR8B2 / B3 CHIKSTQGRSKAFR;

[0108] OR11H1 / 11H2 / 11H12 CSLQNKEIKAALRKR;

[0109] OR13C2 / 13C9 CMKPKSKETLNSDR;

[0110] OR2T5 / 2T29 VNKVSAPECGMQMR;

[0111] OR10H3 / 10H4 CYLKPKGLHSMYSDR;

[0112] OR10K1 / 10K2 CDLLSQKKTISFLR;

[0113] OR1D4 / 5 CQMPSASKKYKTFR;

[0114] OR1D4 / 5 CLQMPSASKKYKTFSR;

[0115] OR3A2 / 3 CRLLSHKSTI...

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Abstract

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. A G protein-coupled receptor antibody is prepared by coupling an antigenic determinant polypeptide with hemocyanin to form a whole antigen, and then using polyclonal antibody technology; the antigenic determinant polypeptide includes multiple polypeptide immunogens, and the The polypeptide immunogens are modified with cysteine ​​and arginine, respectively. The G protein-coupled receptor antibody provided by the present invention can recognize G protein-coupled receptors with similar homology at the same time, and the G protein-coupled receptor kit produced by using the antibody is aimed at cellular level G protein-coupled receptors Signal fast detection.

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 largest 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(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