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Recombinant humanized anti-hepatitis B surface antigen (HBs Ag) Fab antibody and its preparaticon method

A human-derived antibody technology, applied in the field of human-derived anti-HBsAg Fab antibody and its preparation, antibody and its preparation, can solve the problems of small expression and low purity of antibody

Inactive Publication Date: 2006-06-28
余宙耀 +1
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Problems solved by technology

However, its expression system is not a eukaryotic expression system, and the expressed protein still has a certain distance from the natural state; the expression amount is relatively small; the antibody purity is low

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  • Recombinant humanized anti-hepatitis B surface antigen (HBs Ag) Fab antibody and its preparaticon method
  • Recombinant humanized anti-hepatitis B surface antigen (HBs Ag) Fab antibody and its preparaticon method
  • Recombinant humanized anti-hepatitis B surface antigen (HBs Ag) Fab antibody and its preparaticon method

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Embodiment Construction

[0025] 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.

[0026] 1. Phage display technology to obtain anti-HBs Fab antibody fragment gene

[0027] 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. Segment genes were inserted into corresponding sites of the vector pComb3H (Carlos F.Barbas and Dennis R.Burton, 1994, ColdSpring Harbor Laboratory, Monoclonal Antibodies from combinatorial Libraries) to construct vector plasmids containing antibody light chain and heavy chain Fd segments. Use this plasmid to transform XL1-Blue (CarlosF.Bar...

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Abstract

The present invention provides a recombinant humanized anti-hepatitis B surface antigen (HBsAg) Fab antibody as human hepatitis B surface antibody and its preparation method. The preparation method includes the following steps: utilizing phage display technique to obtain the gene of a humanized anti-HBsAg Fab fragment, making expression in prokaryotic expression system, on this basis utilizing PCK technique to amplifying the light-heavy chain gene of Fab, constructing it into the methyl alcohol yeast expression vector, using two-step process to convert yeast, constructing Fab engineering bacterium, secreting, expressing and producing Fab antbody fragment. The Fab antibody has strong HBsAG combination activity and antigen specificity, and the expression product of said gene has high application value in clinical therapy of hepatic disease.

Description

technical field [0001] The invention relates to an antibody and a preparation method thereof, in particular to a human-derived anti-HBsAg Fab 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 prevention and treatment of hepatitis B. [0003] The bloo...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12P19/34C12P21/00C12N15/63C07H21/04
Inventor 余宙耀粟宽源任向荣高辉杨安钢
Owner 余宙耀
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