Martial-Gros A, Sarin K K, Mukhopadhyay A, Ghosh S
National Institute of Immunology, Aruna Asaf Ali Marg, New-Delhi, India.
J Biotechnol. 1999 Jan 22;67(2-3):205-16. doi: 10.1016/s0168-1656(98)00182-5.
The feasibility of large scale production of human anti-tetanus toxoid monoclonal antibody for therapeutic use was evaluated using a human heterohybridoma. The effects of duration of subculture, transition from static to agitated culture conditions and the level of serum concentration were studied. The level of antibody secreted by the clone decreased with increasing length of subculture and decreasing serum concentration. The clone exhibited heterogeneity in expression of surface IgG after 2 or 7 weeks of subculture in static culture conditions irrespective of the serum concentration. However, a prolonged duration of subculture (9 weeks) in 3% serum medium had an effect on the expression of surface IgG both in static and agitated culture conditions. With respect to total (surface and intracellular) IgG, two distinct cell populations were observed. On long term subculture (9 weeks) in low serum medium (3% FCS), there was a decrease in the population which was the high synthesizer. In addition, when these cells were cultivated in agitated spinner flasks, a defect in secretion of antibodies was observed. Thus a general fall in the amount of antibody in the supernatant of agitated cultures was due to decrease in antibody synthesis as well as the defect in secretion of antibodies.
使用人异源杂交瘤评估了大规模生产用于治疗的人抗破伤风类毒素单克隆抗体的可行性。研究了传代培养时间、从静态培养条件过渡到搅拌培养条件以及血清浓度水平的影响。随着传代培养时间的延长和血清浓度的降低，克隆分泌的抗体水平下降。在静态培养条件下，无论血清浓度如何，传代培养2周或7周后，克隆在表面IgG表达上表现出异质性。然而，在3%血清培养基中延长传代培养时间（9周）对静态和搅拌培养条件下的表面IgG表达均有影响。就总（表面和细胞内）IgG而言，观察到两个不同的细胞群体。在低血清培养基（3%胎牛血清）中进行长期传代培养（9周）时，高合成细胞群体数量减少。此外，当这些细胞在搅拌式转瓶中培养时，观察到抗体分泌缺陷。因此，搅拌培养上清液中抗体量的普遍下降是由于抗体合成减少以及抗体分泌缺陷所致。
