Method for preparing antibodies from human peripheral blood memory B lymphocytes
A technology of B lymphocytes and human peripheral blood, applied in chemical instruments and methods, from serum immunoglobulins, peptides, etc., can solve the problems of long cycle and high cost of antigenic preparation, and achieve the effect of low cost
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Embodiment 1
[0022] Embodiment 1: Preparation of anti-rotavirus antibody
[0023] (1) Using a commercial kit (HumanMemoryBCellIsolationKit, MiltenyiBiotec), isolate memory B lymphocytes from the peripheral blood of volunteers inoculated with rotavirus, and place them in RPMI1640 culture medium containing 10% bovine serum in a 37°C carbon dioxide incubator nourish;
[0024] (2) Add an activator to the culture medium of step (1), and continue culturing for 5 days to differentiate the memory B lymphocytes into plasma cells; the activator consists of: 10 μg / ml R848, 20 ng / ml IL-2 and 10 ng / mlIL-4;
[0025] (3) Resuscitating and culturing human myeloma cell lines, cultured in RPMI1640 culture medium containing 10% bovine serum in an incubator at 37°C and 5% (V / V) carbon dioxide, as figure 1 shown;
[0026] (4) The plasma cells prepared in step (2) are fused with the recovered human myeloma cell lines in step (3) to prepare hybridoma cell lines, and the hybridoma cell lines that can secrete ...
Embodiment 2
[0040] Embodiment 2: Preparation of antinuclear antibody
[0041] (1) Using a commercial kit (HumanMemoryBCellIsolationKit, MiltenyiBiotec), isolate memory B lymphocytes from the peripheral blood of patients with systemic lupus erythematosus, in RPMI1640 culture medium containing 15% bovine serum, at 37 ° C, 5% (V / V) cultivate in the incubator of carbon dioxide;
[0042] (2) Add an activator to the culture medium of step (1), and continue culturing for 5 days to make it differentiate into plasma cells; the activator consists of: 5 μg / ml R848, 10 ng / ml IL-2 and 20 ng / ml IL-4 ;
[0043] (3) Resuscitating and culturing the human myeloma cell line, in DMEM culture medium containing 10% bovine serum, culturing in an incubator at 37°C and 5% (V / V) carbon dioxide;
[0044] (4) The plasma cells prepared in step (2) are fused with the recovered human myeloma cell lines in step (3) to prepare hybridoma cell lines, and the fusion myeloma cell lines that can secrete antinuclear antibo...
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