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Preparation method of IL-4R resistant single-chain antibody, and application of IL-4R resistant single-chain antibody to tumor resistance

A single-chain antibody and bacteriophage technology, applied in the field of biomedicine, can solve the problems of low immunity, expensive surgery, radiotherapy and chemotherapy, and loss of qi and blood.

Inactive Publication Date: 2016-07-13
GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

At present, surgical resection, radiotherapy and chemotherapy are still the main means of treating tumors. However, these treatments are likely to cause various side effects such as blood loss and low immunity, and the cost of surgery, radiotherapy and chemotherapy has become economical for many people. burden
Therefore, finding high-efficiency and low-toxic traditional Chinese medicine to treat tumors has become the goal pursued by medical scientists at home and abroad, but there are very few drugs that can really achieve the goal of high efficiency and low toxicity.

Method used

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  • Preparation method of IL-4R resistant single-chain antibody, and application of IL-4R resistant single-chain antibody to tumor resistance
  • Preparation method of IL-4R resistant single-chain antibody, and application of IL-4R resistant single-chain antibody to tumor resistance
  • Preparation method of IL-4R resistant single-chain antibody, and application of IL-4R resistant single-chain antibody to tumor resistance

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Embodiment 1

[0032] 1. Preparation of anti-IL-4R single chain antibody

[0033] 1 material

[0034] 1.1 Main reagents and equipment: PCR instrument was purchased from Bio-Rad Company of the United States; biochemical incubator was purchased from Guangdong Medical Instrument Factory, full-wavelength microplate reader, gel imager\RevertAidFirstcDNASynthesis kit, Mval (BstNI) were purchased from Thermo Company. rh-IL-4R protein was purchased from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd.; Freund's incomplete adjuvant and Freund's complete adjuvant were purchased from Sigma; RNAisoPlus was purchased from TaKaRa; fast agarose gel DNA recovery kit Ⅱ. High-purity plasmid miniprep kit and 2×PowerTaqPCR MasterMix were purchased from Beijing Biotech Co., Ltd.; DNAMarker, ampicillin, kanamycin, PEG8000, and nalidixic acid were purchased from Beijing Suolaibao Biotechnology Co., Ltd. Co., Ltd.; NotⅠ, SfiⅠ, TransStartFastPfuDNA polymerase were purchased from Beijing Quanshijin Biotechnology Co.,...

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Abstract

The invention discloses a preparation method of an IL-4R resistant single-chain antibody and application of the IL-4R resistant single-chain antibody to tumor resistance. The preparation method comprises the following steps: performing animal immunization, constructing pCANTAB5E-scFv recombinant plasmids, constructing IL-4R resistant antibody libraries, enriching and screening phage antibody libraries, optimizing scFv sequences, constructing pET-32a-scFv recombinant plasmids, performing enzyme digestion and identification on pET-32a-scFv recombinant plasmids, expressing cFv recombination protein, performing SDS-PAGE analysis, performing renaturation and purification on expression protein, performing Western blot analysis and the like. According to the invention, the stable and specific preparation method of the IL-4R single-chain antibody is established and can be used for intervening lung cancers in vivo and in vitro through purification protein, a new experiment basis is provided for treating the lung cancer, and a favorable foundation is laid for further developing of a novel antitumor drug.

Description

technical field [0001] The invention relates to a preparation method of an anti-IL-4R single-chain antibody and its application in anti-tumor, belonging to the technical field of biomedicine. Background technique [0002] Lung cancer is the malignant tumor with the highest morbidity and mortality, which poses a great threat to human life and health. It is the first cause of death from malignant tumors, and its incidence is still on the rise. At present, surgical resection, radiotherapy and chemotherapy are still the main means of treating tumors. However, these treatments are likely to cause various side effects such as blood loss and low immunity, and the cost of surgery, radiotherapy and chemotherapy has become economical for many people. burden. Therefore, finding high-efficiency and low-toxic traditional Chinese medicine to treat tumors has become the goal pursued by medical scientists at home and abroad, but there are very few drugs that can really achieve the goal of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28A61K39/395A61P35/00
CPCC07K16/2866A61K2039/505A61K2039/542C07K2317/622
Inventor 何光志杨光勇郭海涛刘茜明田维毅蔡琨王文佳梁建东王平韩洁
Owner GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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