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Binding protein aiming at CYFRA21-1, application thereof, tumour diagnosis reagent and kit

A technology for binding proteins and CDR-VH2, applied in the field of antibodies

Active Publication Date: 2021-06-04
DONGGUAN PENGZHI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are some defects in the sensitivity, specificity and affinity of monoclonal antibodies used to detect CYFRA21-1 in China, and there is still a lot of room for improvement. Therefore, this There is still a strong demand for monoclonal antibodies to detect CYFRA21-1 in the field

Method used

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  • Binding protein aiming at CYFRA21-1, application thereof, tumour diagnosis reagent and kit
  • Binding protein aiming at CYFRA21-1, application thereof, tumour diagnosis reagent and kit
  • Binding protein aiming at CYFRA21-1, application thereof, tumour diagnosis reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] In this example, the restriction endonuclease and Prime Star DNA polymerase were purchased from Takara Company. MagExtractor-RNA extraction kit was purchased from TOYOBO Company. BD SMART TM RACE cDNA Amplification Kit was purchased from Takara Company. The pMD-18T vector was purchased from Takara Company. Plasmid extraction kit was purchased from Tiangen Company. Primer synthesis and gene sequencing were performed by Invitrogen.

[0127] This example provides a method for preparing a recombinant antibody against CYFRA21-1

[0128] 1 Construction of recombinant plasmids

[0129] (1) Primer

[0130] Amplify Heavy Chain and Light Chain 5'RACE Primers:

[0131]

[0132] (2) Antibody variable region gene cloning and sequencing

[0133] RNA was extracted from hybridoma cell lines secreting anti-CYFRA21-1 monoclonal antibody, and the first-strand cDNA was synthesized with SMARTERTMRACE cDNA Amplification Kit kit and SMARTER II A Oligonucleotide and 5'-CDS primers ...

Embodiment 2

[0155] Antibody performance testing

[0156] (1) Activity detection of the antibody and its mutants in Example 1

[0157] Further analysis, the heavy chain variable region of the CYFRA21-1 monoclonal antibody (WT) of Example 1 is shown in SEQ ID NO: 13, wherein the amino acid sequence of each complementarity determining region of the heavy chain is as follows:

[0158] CDR-VH1: G-F-S(X1)-F-S-D(X2)-F(X3)-G-M-H;

[0159] CDR-VH2: Y-L(X1)-S-S-G-S-D-T(X2)-V(X3)-Y-Y-A-D-T(X4)-V-K-G;

[0160] CDR-VH3: A-K(X1)-S-N(X2)-IL(X3)-N-W-F-A;

[0161] Its light chain variable region is shown in SEQ ID NO: 11, wherein the amino acid sequence of each complementarity determining region of the light chain is as follows:

[0162] CDR-VL1: R-S-S-I(X1)-G-A-I(X2)-T-T-S-K(X3)-Y-A-N;

[0163] CDR-VL2: G-T-Q(X1)-N-R-G(X2)-P;

[0164] CDR-VL3: G-V(X1)-W-F(X2)-S-T(X3)-H-W.

[0165] On the basis of the CYFRA21-1 monoclonal antibody in Example 1, the sites related to antibody activity in the complemen...

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Abstract

The invention discloses a binding protein aiming at CYFRA21-1, application thereof, a tumour diagnosis reagent and a kit, and relates to the technical field of antibodies; the binding protein disclosed by the invention comprises an antigen binding structural domain; and the antigen binding structural domain comprises a complementarity determining region. The binding protein disclosed by the invention can be specifically bound with the CYFRA21-1, has better binding activity and affinity, can be used for detecting the CYFRA21-1 and can also be used for auxiliary diagnosis of tumours taking the CYFRA21-1 as a marker; and more protein choices are provided for detection of the CYFRA21-1 and diagnosis of related tumours.

Description

technical field [0001] The invention relates to the technical field of antibodies, in particular to a binding protein against CYFRA21-1, its application, a tumor diagnostic reagent and a kit. Background technique [0002] Cytokeratin (cytokeratin, CYK) is the intermediate filament of the cell body, the main structural protein that forms the epithelial cytoskeleton, and has the function of maintaining the integrity of epithelial cells. According to the isoelectric point and molecular weight, CYK can be divided into type I and type II. Type I belongs to acidic protein, including CYK-9~CYK-23, and type II belongs to basic protein, including CYK-1~CYK-8. Cytokeratin 19 is a class I acidic protein with a molecular weight of about 40KD, and is the smallest protein in the keratin family. Widely distributed on the surface of normal tissues, such as stratified epithelium and squamous epithelium, and single-layer epithelial cells, such as acini, sweat glands, mammary ducts, trachea, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30G01N33/68G01N33/574
CPCC07K16/30G01N33/68G01N33/57423G01N33/57484C07K2317/565C07K2317/569C07K2317/54C07K2317/55C07K2317/622C07K2317/92
Inventor 崔鹏何志强孟媛钟冬梅马秋燕梁碧游辉
Owner DONGGUAN PENGZHI BIOTECH CO LTD
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