caa a22 4500 606213112 CHVBK 20210128101046.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s10295-014-1570-9 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1570-9 Expression of VH-linker-VL orientation-dependent single-chain Fv antibody fragment derived from hybridoma 2E6 against aflatoxin B1 in Escherichia coli [Elektronische Daten] [Aiping Liu, Yang Ye, Weifeng Chen, Xiaohong Wang, Fusheng Chen] Aflatoxin B1 (AFB1) is a toxic secondary metabolic product, which threatens human and animal health. Antibody is a key factor for immunoassay against toxic stuff like AFB1, and single-chain Fv antibody fragment (scFv) has become a popular format of genetically engineered antibody. In this study, four hybridoma cell lines against AFB1 were obtained, and then scFvs 2E6 derived from hybridoma cell line 2E6 were constructed in different VH/VL orientations. Subsequently, scFvs 2E6 were expressed in E. coli BL21(DE3) mainly in the form of inclusion body. SDS-PAGE, Western blot and ELISA were employed to characterize scFvs 2E6. The results revealed that the yield of inclusion body of scFvs 2E6 in either VH/VL orientation was similar; however, only the scFv in VH-linker-VL orientation showed anti-AFB1 bioactivity after refolding. The present study underscores the importance of choosing optimal VH/VL orientation for scFv construction, and scFv may be favorable for immunoassays in food industry. Society for Industrial Microbiology and Biotechnology, 2014 Aflatoxin B1 nationallicence scFv nationallicence Biotechnology nationallicence Hybridoma nationallicence ELISA nationallicence Food safety nationallicence Liu Aiping College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut Ye Yang College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut Chen Weifeng College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut Wang Xiaohong Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut Chen Fusheng Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 255-262 1367-5435 42:2<255 2015 42 10295 https://doi.org/10.1007/s10295-014-1570-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-014-1570-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Aiping College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut NATIONALLICENCE 700 1- Ye Yang College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut NATIONALLICENCE 700 1- Chen Weifeng College of Food Science and Technology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut NATIONALLICENCE 700 1- Wang Xiaohong Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut NATIONALLICENCE 700 1- Chen Fusheng Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, 430070, Wuhan, Hubei, People's Republic of China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 255-262 1367-5435 42:2<255 2015 42 10295