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
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[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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