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Anti-CA50 antibody and applications thereof

An antigen, CDR-VH1 technology, applied in the fields of biotechnology and medicine, can solve the problems of sensitivity and specificity defects, and achieve the effect of binding protein with strong activity and high affinity

Active Publication Date: 2020-04-17
DONGGUAN PENGZHI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the commonly used immunoassay method for CA50 detection requires the use of monoclonal antibodies against CA50. At present, there are very few raw materials for monoclonal antibodies used to detect CA50 in China, and there are defects in sensitivity and specificity, and there is still a large improvement. space

Method used

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  • Anti-CA50 antibody and applications thereof
  • Anti-CA50 antibody and applications thereof
  • Anti-CA50 antibody and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0134] 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. The hybridoma cell line secreting the Anti-CA50 8D10 monoclonal antibody is the existing hybridoma cell line of the applicant, and it is recovered for use.

[0135] 1. Primers

[0136] Amplify Heavy Chain and Light Chain 5'RACE Primers:

[0137] SMARTER II A Oligonucleotide:

[0138] 5'-AAGCAGTGGTATCAACGCAGAGTACXXXXX-3';

[0139] 5'-RACE CDS Primer (5'-CDS): 5'-(T) 25 VN-3'(N=A,C,G,orT; V=A,G,orC);

[0140] Universal Primer A Mix (UPM): 5’-

[0141] CTAATACGACTCACTATAGGGCAAGCAGTGGTATCAACGCAGAGT-3';

[0142] Nested Universal Primer ...

Embodiment 2

[0173] Although the antibody obtained in Example 1 (with sequences such as light chain and heavy chain shown in SEQ ID NO: 11 and 12) has the ability to bind CA50, its affinity and antibody activity are not ideal, so the applicant passed the antibody. The light chain CDRs and heavy chain CDRs were mutated.

[0174] After analysis, the complementarity determining region (WT) of the heavy chain:

[0175] CDR-VH1 is G-I(X1)-T-F-S(X2)-D-A-F(X3)-M-D;

[0176] CDR-VH2 is I-G-Q(X1)-N-G-N-D(X2)-H-A-T-Y-F(X3)-A-E-S-V;

[0177] CDR-VH3 is T-L(X1)-R-F-P(X2)-Y;

[0178] Complementarity-determining regions of the light chain:

[0179] CDR-VL1 is Q-E(X1)-L(X2)-N-S-Y-L(X3)-S;

[0180] CDR-VL2 is R-P(X1)-K-K(X2)-L-I(X3)-D;

[0181] CDR-VL3 is L-N(X1)-Y-D-D(X2)-I(X3)-P-R-T;

[0182] Among them, X1, X2, and X3 are mutation sites.

[0183] Table 1 Mutation sites related to antibody activity

[0184]

[0185]

[0186] After the mutation, the antibody activity was detected, and the c...

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Abstract

The present invention relates to a novel isolated binding protein containing a CA50 antigen binding domain, and researches of the preparation, the applications and the like of the binding protein. Thebinding protein is high in activity, has high affinity with human CA50, and can be widely applied to the field of CA50 detection.

Description

technical field [0001] The invention relates to the fields of biotechnology and medical technology, in particular to an anti-CA50 antibody and its application. Background technique [0002] CA50 is a non-specific broad-spectrum tumor marker (Tumor Marker). It is a sialate and sialoglycoprotein, a chemical class that reflects the presence of tumors. They either do not exist in normal adult tissues but are only found in embryonic tissues, or their content in tumor tissues greatly exceeds that in normal tissues. Their existence or quantitative changes can indicate the nature of tumors, so as to understand the histogenesis and cell differentiation of tumors , cell function, to help tumor diagnosis, classification, prognosis judgment and treatment guidance. [0003] CA50 is a sialic acid ester and sialic acid glycoprotein, which generally does not exist in normal tissues, but CA50 is widely distributed in various tumor tissues such as colon, rectum, lung, pancreas, gallbladder,...

Claims

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Application Information

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IPC IPC(8): C07K16/30C12N15/13C12N15/85G01N33/574
CPCC07K16/30C12N15/85G01N33/57419G01N33/57446G01N33/57438C07K2317/92C07K2317/565C07K2317/569C07K2317/622C07K2317/54C07K2317/55
Inventor 孟媛钟冬梅游辉范凌云
Owner DONGGUAN PENGZHI BIOTECH CO LTD