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Human acid fibroblast growth factor resistant single-chain antibody gene

A technology of fibroblasts and growth factors, applied in the field of genetic engineering, can solve the problems restricting the clinical application of monoclonal antibodies, etc., and achieve the effect of strong penetration ability of solid tumors, easy popularization and application, and low price

Inactive Publication Date: 2006-07-12
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many diseases cannot be treated with monoclonal antibody drugs, which largely restricts the clinical application of monoclonal antibodies

Method used

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  • Human acid fibroblast growth factor resistant single-chain antibody gene

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Effect test

Embodiment Construction

[0054] Embodiments of the invention include the following:

[0055] 1. Preparation of anti-human aFGF hybridoma cell line

[0056] 30ug of recombinant human aFGF protein was used to immunize 8-week-old female Balb / c mice. On the 14th and 35th days, the same amount of antigen was immunized respectively. On the 45th day, the mouse tail blood was detected by indirect ELISA. Booster immunization was carried out on day 56, and splenocytes were taken for fusion after 3 days. Complete Freund's adjuvant was used for the first immunization, and incomplete adjuvant was used for the rest.

[0057] The immunized mouse splenocytes and mouse myeloma cells NS-1 were mixed at a ratio of 5:1, centrifuged at 1500 rpm for 5 min, the supernatant was discarded, and placed in a water bath at 37°C. Add 1ml of pre-warmed PEG slowly, shake gently while adding, and then dilute with 50ml of pre-warmed RPMI1640 incomplete medium. Centrifuge at 800rpm for 5min, discard the supernatant, add enough HAT me...

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PUM

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Abstract

The invention relates to a single-chain antibody gene, recombinant vector and expressing product of anti-human acid desmocyte growth factor in the field of gene project technology. It provides a single-chain Fv which has real value. ScFv is a signal chain which is formed by two variable areas of a light chain and a heavy chain. It folds the two variable areas whose non-covalent bond forms the section with antigen conjugated function.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a single-chain antibody gene of anti-human acidic fibroblast growth factor, its recombinant vector and expression product. Background technique [0002] Acid fibroblast growth factor (aFGF) is a member of the fibroblast growth factor family, which can affect the growth, differentiation and function of various cells such as mesenchymal cells, endocrine cells and nerve cells. Participate in the process of growth and development and tissue damage repair in physiological and pathological processes. [0003] Recent studies have shown that aFGF gene mRNA is overexpressed in transitional cell carcinoma of the bladder, ovarian cancer and ovarian borderline tumors, compared with normal ovaries, ovarian tumor-like lesions and benign ovarian tumors, there are significant differences. When cancer occurs, due to the action of carcinogenic factors, the activity of aFGF...

Claims

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Application Information

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IPC IPC(8): C12N15/13C07K16/18
Inventor 朱筱娟王兴智杨涛
Owner NORTHEAST NORMAL UNIVERSITY
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