[Expression and optimization of anti-AFB1 scFv in Escherichia colil].
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.


CONCLUSIONS
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.
Yang
Lian
L
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China. yanglian27@yahoo.com.cn
Zhang
Yanhong
Y
Ding
Husheng
H
Wang
Liyun
L
Chen
Wei
W
Zhang
Hao
H
D016604:Aflatoxin B1 / Q000235:genetics / Q000276:immunology*; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D046508:Culture Techniques; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D002479:Inclusion Bodies / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D016604:Aflatoxin B1