Monoclonal Antibodies Division, Havana University, Cuba,
Cytotechnology. 2001 Mar;35(2):145-54. doi: 10.1023/A:1017921702775.
The protein-free medium TurboDoma HP.1 (THP.1) was used to produce the CB.Hep-1 monoclonal antibody (mAb) in a CP-1000 hollow fiber bioreactor (HFB). This mAb is used for the immunopurification of recombinant hepatitis B surface antigen (rHBsAg), which is included in a vaccine preparation against the Hepatitis B Virus. By using the experimental conditions tested in this work we were able to generate more than 433 mg of IgG in 43 days. The maximum antibody concentration obtained was about 2.4 mg ml(-1)and the IgG production per day was approximately 11 mg of monoclonal antibody, which constitutes a good concentration value in comparison to the results obtained in ascitic fluid, where concentration for this hybridoma was around 3 mg ml(-1). We used different analytical methods to control the quality of mAbs, obtained from the in vitro system. They included affinity constant determination, analysis of N-glycan structures, immunoaffinity chromatography and antigen binding properties. The results obtained suggest that no significant changes occurred in the mean characteristics of the mAb harvested from the bioreactor during the 43 days of cultivation.
无蛋白培养基 TurboDoma HP.1（THP.1）用于在 CP-1000 中空纤维生物反应器（HFB）中生产 CB.Hep-1 单克隆抗体（mAb）。该 mAb 用于免疫纯化重组乙型肝炎表面抗原（rHBsAg），该抗原包含在乙型肝炎病毒疫苗制备中。通过使用本工作中测试的实验条件，我们能够在 43 天内产生超过 433mg 的 IgG。获得的最大抗体浓度约为 2.4mg/ml，每天产生的 IgG 约为 11mg 单克隆抗体，与腹水获得的结果相比，这是一个很好的浓度值，在腹水中，该杂交瘤的浓度约为 3mg/ml。我们使用了不同的分析方法来控制从体外系统获得的 mAb 的质量。它们包括亲和常数测定、N-糖链结构分析、免疫亲和层析和抗原结合特性。结果表明，在 43 天的培养过程中，从生物反应器中收获的 mAb 的平均特性没有发生显著变化。
