Improvement in the binding specificity of anti-isomiroestrol antibodies by expression as fragments under oxidizing conditions inside the SHuffle T7 E. coli cytoplasm
Issued Date
2022-10-01
Resource Type
ISSN
09168451
eISSN
13476947
Scopus ID
2-s2.0-85138491396
Pubmed ID
35876636
Journal Title
Bioscience, Biotechnology and Biochemistry
Volume
86
Issue
10
Start Page
1368
End Page
1377
Rights Holder(s)
SCOPUS
Bibliographic Citation
Bioscience, Biotechnology and Biochemistry Vol.86 No.10 (2022) , 1368-1377
Suggested Citation
Juengsanguanpornsuk W., Kitisripanya T., Boonsnongcheep P., Yusakul G., Srisongkram T., Sakamoto S., Putalun W. Improvement in the binding specificity of anti-isomiroestrol antibodies by expression as fragments under oxidizing conditions inside the SHuffle T7 E. coli cytoplasm. Bioscience, Biotechnology and Biochemistry Vol.86 No.10 (2022) , 1368-1377. 1377. doi:10.1093/bbb/zbac126 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/85496
Title
Improvement in the binding specificity of anti-isomiroestrol antibodies by expression as fragments under oxidizing conditions inside the SHuffle T7 E. coli cytoplasm
Other Contributor(s)
Abstract
Sensitive and specific analysis of isomiroestrol (Iso) is required for the quality control of Pueraria candollei, a herb used to treat menopausal disorders. The anti-isomiroestrol monoclonal antibody (Iso-mAb) exhibits cross-reactivity with miroestrol and deoxymiroestrol, which impacts the analytical results. Here, the active and soluble forms of the single-chain variable fragment (Iso-scFv) and fragment antigen-binding (Iso-Fab) against Iso were expressed using Escherichia coli SHuffle® T7 to alter the binding specificity. The Iso-scFv format exhibited a higher binding activity than the Iso-Fab format. The reactivity of Iso-scFv towards Iso was comparable with that of the parental Iso-mAb. Remarkably, the binding specificity of the scFv structure was improved and cross-reactivity against analogs was reduced from 13.3-21.0% to 1%. The structure of recombinant antibodies affects the binding characteristics. Therefore, the immunoassays should improve specificity; these findings can be useful in agricultural processes and for quality monitoring of P. candollei-related materials.