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Updated: Jun 19, 2026
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
Published on: October 15, 2016
[Expression and optimization of anti-AFB1 scFv in Escherichia colil]
Lian Yang1, Yanhong Zhang, Husheng Ding
1State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China. yanglian27@yahoo.com.cn
Objective:
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.
Methods:
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.
Results:
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.
Conclusion:
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.
