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Recombinant human antibody against HBsAg and its preparation method

A surface antigen, human-derived technology, applied in recombinant DNA technology, anti-animal/human immunoglobulin, using vectors to introduce foreign genetic material, etc., can solve the problems of difficult renaturation and low expression

Inactive Publication Date: 2005-01-26
余宙耀 +2
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Problems solved by technology

[0004] For the research of genetically engineered human hepatitis B surface antibody, phage display library technology is used at home and abroad to obtain its Fab segment (fragment of antigen binding) gene, and then expressed in Escherichia coli, but the problem is that the Fab antibody is double-chain , secreted expression in Escherichia coli, the expression level is very low, and it is difficult to refold when expressed in inclusion bodies

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  • Recombinant human antibody against HBsAg and its preparation method
  • Recombinant human antibody against HBsAg and its preparation method
  • Recombinant human antibody against HBsAg and its preparation method

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Embodiment Construction

[0060] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the embodiments are only used to illustrate the present invention rather than limit the present invention.

[0061] 1. Obtaining anti-HBsAg Fab antibody fragment gene by phage display technology

[0062] On the fifth day after the booster injection of hepatitis B vaccine volunteers, blood was collected, monocytes were isolated, mRNA was extracted, cDNA was synthesized by reverse transcription, and the light chain gene and heavy chain Fd of the anti-HBsAg antibody were amplified by PCR with the designed primers. The fragment genes were respectively inserted into the corresponding sites of the vector pComb3H (Carlos F.Barbas and Dennis R.Burton, 1994, Cold Spring Harbor Laboratory, Monoclonal Antibodies from combinatorial Libraries) to construct vector plasmids containing antibody light chain and heavy chain Fd segments. Transform the competent...

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Abstract

The present invention discloses a recombinant human antibody against HBsAg single-chain antibody used as human hepatitis B surface antibody and its preparation method. The Fab segment gene for cloning the human HBsAg antibody by bacterial virus display technology is expressed in bacillus coli; The HBsAg single-chain antibody gene can be obtained by superposing PCR and the gene is inserted into a pronucleus cytoplasm expression carrier, the high efficiency expression can be obtained in the bacillus coli, then the recombinant single-chain antibody is purified and recovered to original property; the recombinant human antibody against HBsAg single-chain antibody used as human hepatitis B surface antibody has an activity combined with the HBsAg, and the primary structure of its protein is quite correct by measuring mass spectra. The single-chain antibody constructed in the invention can replace the blood source antibody, and can be used for passive immunity of hepatitis B and can be developed for guide medicine having curing function based on it.

Description

technical field [0001] The invention relates to an antibody and a preparation method thereof, in particular to a recombinant human-derived anti-hepatitis B surface antigen (HBsAg) single-chain antibody and a preparation method thereof, belonging to the field of biological products. Background technique [0002] Hepatitis B is a worldwide epidemic infectious disease. There are more than 300 million hepatitis B virus carriers in the world, among which there are more than 100 million hepatitis B virus carriers in my country, and more than 10 million hepatitis B patients. According to the statistics of the World Health Organization, more than one million people die from this disease every year all over the world, and there is no effective treatment method in clinical practice. Since there is no effective means to treat hepatitis B at present, it is particularly important to prevent its infection in controlling the infection of the disease, making prevention the focus of preventi...

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Application Information

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IPC IPC(8): C07K16/18C12N15/63C12N15/70C12P21/00
Inventor 余宙耀粟宽源任向荣高辉杨安钢
Owner 余宙耀
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