C50株抗CEA杂交瘤细胞抗体生成动力学及体外培养条件的优化

Production Kinetics of Antibody to C50 Strain of CEA Hybridoma Cell Line and Optimization of In Vitro Culture Condition

目的 研究C5 0株抗CEA杂交瘤细胞的抗体生成动力学 ,确定优化体外培养条件。方法 体外静态培养C5 0细胞 ,采用ELISA和细胞计数的方法检测培养上清抗体浓度和细胞数量 ,分析细胞生长与抗体分泌的关联性。采用体内复壮和添加新型免疫增强剂CpGODN的方法 ,尝试恢复长期体外培养的C5 0细胞的抗体生成能力。结果 C5 0细胞的抗体生成动力学为非生长关联型 ,经过体内复壮 ,C5 0细胞体外培养上清中抗体浓度显著提高 ,但活细胞密度或细胞活力均未发生明显变化。在培养基中添加一定浓度的CpGODN ,可以恢复长期体外培养的C5 0细胞的抗体生成能力。结论 根据细胞的抗体生成动力学特征 ,通过调节细胞的生长状态 ,可提高上清抗体浓度。根据抗体基因的组织特异性表达的机制 ,通过优化杂交瘤细胞的体外培养条件 ,能保持细胞正常的抗体生成能力 。

Objective To study the production kinetics of antibody to C50 strain of CEA hybridoma cell line and determine the optimal in vitro culture condition of the cell line.Methods Culture C50 cells in vitro, detect the antibody concentration in culture supernatant by ELISA,count the cells and analyze the relationship between cell growth and antibody secretion.On the basis of this,the authors attempted to recover the antibody producing ability of C50 cells after long term in vitro culture by rejuvenating test in vivo and adding a novel immunopotentiator CpG ODN.Results The production kinetics of antibody to C50 cell was non growth related.After rejuvenating in vivo,the antibody concentration in supernatant of in vitro culture of C50 cells increased significantly.However,no significant change was observed in live cell density or cell viability.The addition of a certain concentration of CpG ODN could recover the antibody producing ability of C50 cells after long term in vitro culture.Conclusion The antibody concentration in culture supernatant can be increased by adjusting the growth of C50 cells.The normal antibody producing ability of C50 cells,as well as high and stable antibody concentration in culture supernatant,can be maintained by optimizing the in vitro culture condition of CEA hybridoma cells.