近日，國際著名雜誌Molecular Biotechnology 在線刊登了上海巴斯德研究所周保羅研究組的最新研究成果「High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreactor，」這是首次關於用波浪生物反應器灌注培養穩轉了人單克隆抗體基因的果蠅S2細胞來產生人單克隆抗體的報導。
治療性人單克隆抗體已經增長為數十億美元的產業，因而建立一個穩定的培養條件以滿足人單克隆抗體的大規模表達是非常重要的。波浪生物反應器於二十世紀九十年代後期被用於哺乳動物及昆蟲細胞的蛋白表達，但是用波浪生物反應器來灌注培養穩轉了人單克隆抗體基因的果蠅穩轉細胞表達人單克隆抗體蛋白尚屬首次。
在該研究中上海巴斯德所的研究助理王璐嵐和博士生胡紅星在周保羅教授的指導下克隆了抗高致病性禽流感病毒的血凝蛋白的人單克隆抗體基因，並構建到果蠅誘導型表達載體裡面，穩轉了果蠅S2細胞系，通過有限稀釋法拿到高表達人單克隆抗體的單細胞克隆。對穩轉了人單克隆抗體的單克隆細胞株分別用來在灌注和非灌注的生物反應器中進行比較培養，發現灌注培養方法在獲得細胞數量與表達抗體蛋白產量上明顯優於後者，且這兩種方法生產的抗體都是有中和活性的。這些結果表明用波浪生物反應器進行灌注培養S2穩轉細胞系進行大規模蛋白表達是非常好的方法。
該研究是與GE公司研發部Fast Trak中心的楊建軍博士合作完成的，得到了國家自然科學基金及李嘉誠基金會的資助。（生物谷Bioon.com）
High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreactor
Lulan Wang, Hongxing Hu, Jianjun Yang, Feng Wang, Christian Kaisermayer and Paul Zhou
Since it was first introduced in late 1990s Wave bioreactor has been used for protein production by mammalian and insect cell lines. However, using Wave bioreactor to produce human monoclonal antibody by stable Drosophila Schneider 2 (S2) cell transfectants has not been reported before. In this study, S2 cells were co-transfected with an inducible vector expressing human monoclonal antibody heavy and light chains, respectively, specific for hemagglutinin (HA) of H5N1 influenza virus. Stable S2 transfectant clone was selected by limiting dilution assay. Stable S2 transfectant clone that produce the highest amount of human monoclonal antibody was inoculated into two 2-l disposable cellbags, where cell growth and antibody production were compared between batch and perfusion cultures using Wave bioreactor. Here, we report that maximum viable cell density reached 1.06 × 107 cells/ml in batch culture; whereas 1.04 × 108 cells/ml was achieved in perfusion culture. The maximum volumetric antibody productivity in batch culture was 52 mg/l/day; while perfusion culture yielded 1,437 mg/l/day. As a result, the total antibody production was 201 mg in batch culture and 8,212 mg in perfusion culture. The antibody produced by both cultures displays full neutralizing activity. Thus, our results provide strong support for using Wave bioreactor in perfusion culture for a large-scale production of human monoclonal antibody by stable S2 cell transfectants.