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大量制备McAb 时,通常采用的方法有以下两种:(一)体外培养法:在完全培基中培养杂交瘤细胞,其上清液中McAb 的浓度较低,每ml 中仅含10—100 μg,并且其中含有大量小牛血清,难以除掉,使用很不方便。如果在无血清培基中培养,所得McAb 的浓度更低,必须经过浓缩,透析后,方能使用。(二)体内培养法:将杂交瘤细胞接种到事先腹腔注入过降植烷(Pristane)的BALB/C 小鼠腹腔中,可得到含高浓度McAb 的(5—20mg/ml)腹水,每只小鼠可得5—10ml,一般实验即可应用。这已成为生产McAb 的常规方法。在实验中,我们发现产生腹水的小鼠血清中亦含有高浓度的McAb。为了证实这一问题,我们选用抗人IgG 的杂交瘤细胞株21株,接种32只小鼠。制备腹水。在取腹水之前,眼窝放血,分离血清,与同一小鼠的腹水配对,
When a large number of McAb preparation, the commonly used methods are the following two kinds: (a) in vitro culture method: in complete culture medium hybridoma cells, the lower the concentration of McAb in the supernatant, containing only 10-100 per ml μg, and contains a lot of bovine serum, it is difficult to remove, very inconvenient to use. If cultured in serum-free medium, the resulting McAb is less concentrated and must be concentrated and dialyzed before it can be used. (B) In Vivo Culture: Hybridoma cells were inoculated into the peritoneal cavity of BALB / C mice preperitoneally injected with Pristane to obtain (5-20 mg / ml) ascites containing high concentrations of McAbs, each 5-10ml mice, the general experiment can be applied. This has become the conventional method of producing McAbs. In the experiment, we found that ascites also contains high concentrations of McAbs in the serum of mice. To confirm this problem, we selected 21 anti-human IgG hybridoma cell lines and inoculated 32 mice. Ascites is prepared. Before taking ascites, orbital bleeding, serum separation, and the same mouse ascites paired,