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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, applied in the biological field, achieves obvious effects, simple and convenient preparation, and less toxic and side effects

Active Publication Date: 2012-09-05
WEST VAC BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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] Embodiment 1 The preparation of the fusion toxin of the present invention

[0024] The heavy chain and light chain variable regions of the antibody were amplified from known anti-EGFR hybridomas to construct anti-EGFR single-chain antibodies, and ribosome display technology was used for specificity identification and affinity evolution. Gelonin and the obtained single-chain antibody gene were constructed as a fusion gene, and then expressed in Escherichia coli.

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

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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. The technical 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 fused with Gelonin recombinant immunotoxin and its preparation method and application. Background technique [0002] The epidermal growth factor receptor (EGFR) family is one of the most representative molecules among transmembrane receptor tyrosine kinases, which play 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 tumors derived from various epithelial cells 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 the characteristics that it plays an important role in the growth and differentiation of tumor cells make EGFR a promising target for tumor therapy. [0003] The toxin protein Gelonin (Ge...

Claims

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Application Information

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