Yang Lian, Zhang Yanhong, Ding Husheng, Wang Liyun, Chen Wei, Zhang Hao
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
Wei Sheng Wu Xue Bao. 2009 Jul;49(7):880-8.
A drawback of the expression of single chain antibody fragment (scFv) in prokaryotic system is the protein accumulation in the cytoplasm as inclusion body. We aimed at high-level production of an anti-aflatoxin B1 scFv in functional form.
The gene of scFv-H4 was cloned into pET22b vector and transformed into E. coli BL21(DE3) and Origami (DE3),respectively. The amount of functional scFv-H4 was optimized in terms of IPTG concentration and induction temperature.
scFv-H4 could be expressed in both BL21(DE3) and Origami (DE3). Compared with BL21(DE3), Origami(DE3) could express multifunctional scFv-H4 (35 mg/mL) and less in inclusion body (11% of the total expression). The expression of scFv-H4 was significantly affected by induction temperature rather than IPTG concentration.
The pET22b could be used for high-level expression of the functional scFv-H4 in Origami (DE3), which has an oxidative cytoplasm. In addition,the induction at low temperature avoided the formation of inclusion body.
单链抗体片段（scFv）在原核系统中表达的一个缺点是蛋白在细胞质中以包涵体形式积累。我们旨在高水平生产功能形式的抗黄曲霉毒素B1 scFv。
将scFv-H4基因克隆到pET22b载体中，分别转化到大肠杆菌BL21(DE3)和Origami(DE3)中。根据异丙基-β-D-硫代半乳糖苷（IPTG）浓度和诱导温度对功能性scFv-H4的产量进行优化。
scFv-H4在BL21(DE3)和Origami(DE3)中均能表达。与BL21(DE3)相比，Origami(DE3)能表达多功能scFv-H4（35 mg/mL），且包涵体较少（占总表达量的11%）。scFv-H4的表达受诱导温度的显著影响，而不是IPTG浓度。
pET22b可用于在具有氧化性细胞质的Origami(DE3)中高水平表达功能性scFv-H4。此外，低温诱导可避免包涵体的形成。
