Establishment of a mass production system of human monoclonal anti-toxin antibodies

1999年，日本和中国的研究人员举行了三次会议，同意在兰州生物制品研究所进行人类单抗毒素的生产，并由日本研究人员通过对培养条件的实验室检查来支持生产。由于杂交瘤细胞在生产抗破伤风单抗方面的滞后性，我们致力于杂交瘤细胞在含5%FCS的培养液中的大规模培养以及从培养物中纯化抗体的方法。该杂交瘤细胞经1.5个月的灌流培养，共获得1.5g抗体，其抗体浓度为7×10~(-3)IU/ml，其中以破伤风类毒素亲和层析和Mono-Q离子交换层析相结合的纯化效果最好。G2和G6最终制剂对100μg免疫球蛋白的中和活性分别为6.2IU和20.4IU。这些制剂作为生物制品的特性目前正在中国的国家药品生物制品检定所进行调查。

In 1999, Japanese and Chinese researchers had three meetings and agreed that the production of human monoclonal antitoxins would be performed in Lanzhou Institute of Biological Product and that Japanese researchers would support the production by laboratory examination of culture conditions. Because of the delay in adaptation of hybridomas for the production of human anti-tetanus monoclonal antibodies, we focused our effort on the mass culture of the hybridomas in the medium containing 5 % FCS and the way of purification of antibodies from the culture. Perfusion culture of the hybridomas for 1.5 months gave us 1.5 g of IgG in total, and the concentration of the antibodies in the culture was 7X10^<-3> IU/ml. The most effective way of purification was proved to be a combination of an affinity chromatography with tetanus toxoid and an ion-exchange chromatography with Mono Q column. Neutralizing activities of final preparations of G2 and G6 were 6.2 IU and 20.4 IU for 100 μg IgG, respectively. Characteristics of these preparations as biological products are now under investigation in National Institute for the Control of Pharmaceuticals and Biological Products in China.