Method for constructing hybridoma cell line of heterogeneity of seereting human monoclonal antibody for anti hepatitis B

A technology of human monoclonal antibody and hybridoma cells, which is applied in the field of construction of heterogeneous hybridoma cell lines, and can solve problems such as imperfect technology and unsatisfactory antibody expression

Inactive Publication Date: 2005-03-30
SHAANXI JIUZHOU BIOTECH
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Problems solved by technology

[0004] At present, the technical development of the development of human monoclonal antibodies in the world is mainly reflected in the development of "human-derived antibodies" by DNA recombination technology. Research in this area has also been carried out in China, but the development of "human-derived"

Method used

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  • Method for constructing hybridoma cell line of heterogeneity of seereting human monoclonal antibody for anti hepatitis B
  • Method for constructing hybridoma cell line of heterogeneity of seereting human monoclonal antibody for anti hepatitis B
  • Method for constructing hybridoma cell line of heterogeneity of seereting human monoclonal antibody for anti hepatitis B

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Experimental program
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Embodiment

[0025] 1. Preparation of various cells:

[0026] 1. Preparation of anti-HBs positive human spleen lymphocytes (SL):

[0027] Spleen donor: A 45-year-old male with a history of chronic HB. He was hospitalized due to splenomegaly and underwent splenectomy. Anti-HBs positive in menstrual blood test, intramuscular injection of Hepatitis B vaccine one week before splenectomy, aseptically take out spleen cortex tissue immediately when spleen is taken out, homogenate spleen tissue to separate splenocytes, wash with RG (serum-free medium) centrifuge 2 times, and then use RG to suspend SL, count live cells by trypan blue exclusion method, and take the required number for later use.

[0028] 2. Murine plasmacytoma cell line P 3 -X 63 Preparation of / AG8-653:

[0029] Take vigorously growing 653 cells and inoculate them in RGS (complete medium) medium at a concentration of 4×10 4 / ml, cultured for 4 days. In cells up to a concentration of 1 x 10 6 / ml, the viability reaches 98%, a...

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Abstract

This invention provides a method for establishing hetero-hybrid tumor engineering cell sequence for producing human monoclonal antibody for anti-hepatitis B virus. The anti-HBs positive human splenic lymph cell (SL) is fused with mouse plasmacytoma sequence P3-X63/Ag8-653 cell. Limiting dilution method is used for cloning the positive antibody poro-hybrid-tumor cell to obtain hetero hybrid tumor engineering cell sequence which produces human monoclonal antibody for anti-hepatitis B virus, with stable and high product, rate. Said monoclonal antibody is proceeded with purification and cross-linkage with interferon to produce target prepn. of anti-hepatitis B virus human monoclonal antibody crosslinking with itnerferon, and passive immune prepn. for hepatitis B, and testing agent for hepatitis B.

Description

technical field [0001] The invention relates to a construction method of a heterogeneous hybridoma engineering cell line, in particular to a construction method of a heterogeneous hybridoma cell line secreting anti-hepatitis B human monoclonal antibody. Background technique [0002] Since the successful development of mouse monoclonal antibodies by hybridoma technology, people have developed a wide variety of mouse monoclonal antibodies, which have been widely used and formed a huge high-tech industry. The use of mouse monoclonal antibodies as therapeutic agents in humans is limited due to the fact that they are heterologous proteins that can cause serious adverse reactions. Therefore, since the 1980s, the research on human monoclonal antibodies has been highly valued by the medical and biological community. It is believed that human monoclonal antibodies can strengthen and improve the treatment of infectious diseases, and human serum immunoglobulin as a treatment model, Hi...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P1/16A61P31/12C12N15/02C12P21/08G01N33/576G01N33/577
Inventor 张永忠
Owner SHAANXI JIUZHOU BIOTECH
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