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Anti-EGFR single-chain antibody fusion Gelonin recombination immunotoxin, preparation method and usage thereof

An immunotoxin and single-chain antibody technology, which is applied in the biological field to achieve obvious effects, simple and convenient preparation, and small toxic and side effects.

Active Publication Date: 2010-03-17
WEST VAC BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the recombinant immunotoxin tumor treatment technology plan using the toxin protein Gelonin to target the epidermal growth factor receptor

Method used

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  • Anti-EGFR single-chain antibody fusion Gelonin recombination immunotoxin, preparation method and usage thereof
  • Anti-EGFR single-chain antibody fusion Gelonin recombination immunotoxin, preparation method and usage thereof
  • Anti-EGFR single-chain antibody fusion Gelonin recombination immunotoxin, preparation method and usage thereof

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Embodiment 1

[0023] Example 1 Preparation of the fusion toxin of the present invention

[0024] The heavy and light chain variable regions of antibodies were amplified from known anti-EGFR hybridomas to construct anti-EGFR single-chain antibodies, which were specifically identified and affinity evolved using ribosome display technology. A fusion gene was constructed between Gelonin and the obtained single-chain antibody gene, and then expressed in E. coli.

[0025] Specifically: extract total RNA from anti-EGFR hybridoma cells (preserved in our laboratory), and then amplify the heavy and light chains (VH, VL) of Anti-EGFR by RT-PCR according to the designed library and specific primers Gene, select the band with good amplification effect for gel recovery, and insert into T vector. The ligation product was transformed into DH5a, a large number of single colonies were picked, the plasmid was extracted, and sent for sequencing. The sequencing results were analyzed by Jellyfish software, and...

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Abstract

The invention belongs to the biotechnology field, in particular to recombination immunotoxin of anti-EGFR single-chain antibody fusion Gelonin, a preparation method thereof and the usage thereof. Thetechnical problem to be solved is to provide recombination immunotoxin taking an epidermal growth factor receptor as a target. The recombination immunotoxin is formed by the single-chain antibody of the EGFR and toxin-protein Gelonin fusion expression. The technical solution plays a role in the specific cytocidal effect on tumor by using the targeting ability of the single-chain antibody on the EGFR high- expression tumor and the cytotoxic effect of Gelonin and has obvious effect; the preparation method is simple and convenient, has good application prospect, and provides a new choice for thetreatment of the tumor.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an anti-EGFR single-chain antibody fusion Gelonin recombinant immunotoxin and a preparation method and application thereof. Background technique [0002] Epidermal growth factor receptor (epidermal growth factor receptor, EGFR) family is one of the most representative molecules of transmembrane receptor-type tyrosine kinase, which plays an important role in the process of cell growth, proliferation and differentiation. Studies have shown that there are abnormal activation, amplification and overexpression of EGFR gene in a variety of epithelial cell-derived tumors, such as breast cancer, colorectal cancer, lung cancer, head and neck squamous cell carcinoma, and pancreatic cancer. The high expression of EGFR in tumors and its important role in tumor cell growth and differentiation make EGFR a promising target for tumor therapy. [0003] The toxin protein Gelonin (Gel) is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21A61K38/16A61K48/00A61K47/48A61P35/00C12R1/19A61K47/68
Inventor 李炯魏于全
Owner WEST VAC BIOPHARMA CO LTD
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