人源抗HBsAgFab工程化大肠杆菌优化表达的实验研究.pdfVIP

1022 2003 1128 11 Med J Chin PLA Vol 28 No 11 November 2003 人源抗HBsAg Fab 工程化大肠杆菌优化表达的实验研究 510515 郑大勇 罗荣城 韩焕兴 HBsAg Fab , HBsAg Fab , , , , , 25 02% arabinose 12h Fab , OD600 552, 110g/ L , Fab , ; , Fab; ; ; + R3782 2; R3 2-33 OPTIMIZING EXPRESSION CONDITIONS OF ENGINEERED ESCHERICHIA COLI BEARING HUMANIZED ANTI-HB- sAG Fab ON LABORATORY SCALE ZHENG Da-yong, LUO Rong-cheng, HAN Huan-xing. Nanfang Hospital, First Military Medical University, Guangzhou 501515, China Abstract Object ve To investigate thebest fermentation and induction models of the engineered Escherichia coli, in order to obtain the highest expression level of humanized ant-i HBsAg Fab. MethodsThe optimal condition governing the growth of the Escherichia coli, with the flask-shaking method and the best fermentation and induction conditions to yield the highest production level were explored, and then Ecoliwere grown in fermen- tor using the fed-batch method following the same principle under flask-shaking condition to ensure the best production. Results The data obtained from flask-shaking conditions showed that when the induction procedure started the amount of ant-i HBsAg Fab would reach the highest level at mid- log growth phase under the induction condition of 25 and 02% arabinose. Using the DO-stat fed-batch method, the OD600 value of the culture would reach 552, which corresponded to 110g/ L bacterial wet weight. The biological activity of Fab was proved to have well preserved. Conclus on We established the optimal production technic of HBsAg Fab, and lay a foundation to produceHBsAg Fab on a large scale Key wordsengineered antibody; immunoglobulins, Fab; Escherichia coli; fermentation; optimized expression HBsAg Fab [ tryptone 10g/ L, yeast extract 10g/ L, KH PO 2 4 ,, 2g/ L, K