Moro A M, Rodrigues M T, Gouvea M N, Silvestri M L, Kalil J E, Raw I
Centro de Biotecnologia, Instituto Butantan, Sao Paulo, Brazil.
J Immunol Methods. 1994 Nov 10;176(1):67-77. doi: 10.1016/0022-1759(94)90351-4.
Monoclonal antibodies are one of the most important products of biotechnology and laboratories and companies all over the world are pursuing their large-scale production. Herein we report a protocol for hybridoma cell cultivation over small glass cylinders inside a 3 liter bioreactor vessel which leads to the production and purification--in order of grams--of one MAb intended for human therapeutic use. This protocol proved to be simple, reproducible and cost effective. Three trials are reported: the first two using conventionally serum-supplemented medium culture and producing 3.15 and 2.1 g of purified MAb in 30 and 21 days respectively, and the third one using serum-free medium culture and producing 6 g of purified MAb in 36 days. We have ascertained the stability of the hybridoma by its cloning directly in serum-free medium. The downstream processing of the serum-free trial was done in a single step, concentrating large volumes of supernatant while simultaneously purifying the antibody.
单克隆抗体是生物技术最重要的产品之一，全世界的实验室和公司都在致力于其大规模生产。在此，我们报告一种在3升生物反应器容器内的小玻璃圆筒中培养杂交瘤细胞的方法，该方法可实现用于人类治疗的一种单克隆抗体按克级的生产和纯化。该方法被证明是简单、可重复且具有成本效益的。报告了三项试验：前两项使用传统的含血清培养基培养，分别在30天和21天内生产出3.15克和2.1克纯化单克隆抗体，第三项使用无血清培养基培养，在36天内生产出6克纯化单克隆抗体。我们通过直接在无血清培养基中克隆确定了杂交瘤的稳定性。无血清试验的下游加工在一步中完成，在浓缩大量上清液的同时纯化抗体。
