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epcam single domain antibody e6

A single-domain antibody and expression vector technology, applied in the field of molecular biology, can solve problems such as unsatisfactory clinical trial results of immune-directed therapy

Active Publication Date: 2021-07-23
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to disclose an EpCAM single-domain antibody for the problems of the unsatisfactory clinical trial results of EpCAM immune-directed therapy in the prior art.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1. Construction of EpCAM single-domain antibody library

[0057] (1) Purchasing commercially available EpCAM recombinant proteins (ORIGENE Company) Here, other commercially available EpCAM recombinant proteins can be used, and the way to purchase EpCAM recombinant proteins will not affect the realization of the purpose of subsequent experiments. According to the instructions, the purchased recombinant protein was taken out, mixed with an equal volume of Freund's incomplete adjuvant and fully emulsified, and used as a vaccine to immunize alpacas.

[0058] (2) Using the method of subcutaneous multi-point injection, immunize two adult male alpacas for 5 consecutive immunizations, with a one-week interval between each time, to promote the production of EpCAM antigen-specific single-domain antibodies;

[0059] (3) After 5 times of immunization, according to the detection titer (take 5ml of blood from the common carotid artery before each immunization, collect the upper serum...

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Abstract

The present invention belongs to the field of molecular biology, and specifically relates to EpCAM single domain antibody E6. Using phage display technology, the single domain heavy chain antibody library that can specifically bind to the target antigen EpCAM is screened from the single domain heavy chain antibody library immune to camel origin , and further cloned the gene sequence encoding the antibody into a prokaryotic expression vector to obtain a highly expressed EpCAM single domain antibody. It can significantly improve the specific binding performance of single-domain antibodies, and provide a basis for the clinical application of EpCAM immune-directed therapy.

Description

technical field [0001] The invention relates to an EpCAM single domain antibody and a preparation method thereof, belonging to the field of molecular biology. Background technique [0002] EpCAM refers to epithelial cell adhesion molecule (Epithelial Cell Adhesion Molecule,), also known as CD326, is a kind of glycoprotein widely expressed in human epithelial tissue, and has a very potent antigenic epitope. EpCAM not only has the function of epidermal cell adhesion molecule, but also participates in multiple life activities such as accelerating cell cycle, promoting cell proliferation, differentiation, migration and immune escape. [0003] Studies have shown that EpCAM activates the expression of proto-oncogenes such as c-myc and cyclinA / E by participating in the β-catenin-dependent Wnt cascade reaction, leading to tumorigenesis. [0004] Under physiological conditions, EpCAM is expressed to varying degrees in all normal epithelium except squamous epithelial cells, and most ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/30C12N15/13C12N15/70C12N1/21A61K39/395A61P35/00G01N33/574
CPCA61K39/00A61P35/00C07K16/30C07K2317/569G01N33/57484
Inventor 季晨博
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL