Single-chain antibody KGH-R1-ScFv for resisting p21Ras protein and application thereof

A p21ras, kgh-r1-scfv technology, applied in the field of medical biology, can solve the problems of weak vascular permeability, strong immunogenicity and long half-life

Inactive Publication Date: 2012-11-21
杨举伦
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Due to the large molecular weight, weak vascular permeability, poor ability to diffuse into tumors, long half-life and strong immunogenicity of intact antibodies used for tumor biotherapy, their targeted delivery and tumor antagonism application effects are limited.

Method used

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  • Single-chain antibody KGH-R1-ScFv for resisting p21Ras protein and application thereof
  • Single-chain antibody KGH-R1-ScFv for resisting p21Ras protein and application thereof
  • Single-chain antibody KGH-R1-ScFv for resisting p21Ras protein and application thereof

Examples

Experimental program
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Embodiment 1

[0050] Example 1 : A kind of anti-p21Ras protein single-chain antibody KGH-R1-ScFv and application thereof, the technical scheme of its acquisition is as follows:

[0051] 1. Total RNA extraction and cDNA synthesis of anti-p21Ras hybridoma cell lines

[0052] 1.1 Total RNA extraction

[0053] The hybridoma cells cultured to the logarithmic growth phase were collected (preserved in the China Center for Type Culture Collection on September 23, 2011, with the preservation number CCTCC NO: C201197), using Trizol (obtained from Mrcgene Company), according to the reagent instructions, Total RNA was extracted from hybridoma cells. The integrity of total RNA was detected by 1.0% agarose gel electrophoresis. Total RNA concentration was measured with a NanoDrop 2000 (from Thermo) spectrophotometer.

[0054] 1.2 cDNA synthesis

[0055] Use RevertAid TM H Minus First Strand cDNA Synthesis Kit (obtained from Fermentas Company), according to the instructions of the kit, the total RN...

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Abstract

The invention provides a single-chain antibody KGH-R1-ScFv for resisting p21Ras protein and application of the single-chain antibody KGH-R1-ScFv, belonging to the field of medical biology, and particularly relates to a recombination human adenovirus vector Ad-hrGFP-ScFv with a single-chain antibody coding gene KGH-R1-ScFv. The single-chain antibody provided by the invention can specifically recognise and perform antagonism to p21Ras proteins with different sources, different tissues and different cell strains in a broad-spectrum way, and also can inhibit the proliferation capacity and invasion capacity of human tumor cell of p21Ras protein with high expression in vitro, reduce the ratio of cells in cell division period, and inhibit the growth of transplantation tumor of human tumor cell of p21Ras protein with high expression in mice.

Description

technical field [0001] The invention belongs to the field of medical biology, and in particular relates to an anti-p21Ras protein single-chain antibody KGH-R1-ScFv and application thereof. Background technique [0002] Tumors, especially malignant tumors, seriously endanger human life and health. Carrying out research on the prevention and treatment of malignant tumors has very important practical significance for preventing the occurrence of tumors, reducing the incidence of tumors, reducing mortality, and guiding clinical treatment. [0003] Ras gene is an important proto-oncogene, which is involved in the occurrence and development of various tumors. Its expression product Ras protein is an important molecular switch involved in cell proliferation, growth, differentiation and apoptosis. Studies have shown that ras gene mutation and (or) Ras protein overexpression are closely related to the occurrence and development of tumors, and play a very important role in the patho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/32C12N15/13C12N15/861C12N1/21C12R1/19
Inventor 杨举伦胡奇婵陈玥王丽丁峰
Owner 杨举伦
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