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Enzyme Microb Technol. 2016 Mar;84:50-5. doi: 10.1016/j.enzmictec.2015.12.011. Epub 2015 Dec 24.

Development of high-affinity single chain Fv against foot-and-mouth disease virus.

Author information

1
Department of Chemical and Biomolecular Engineering, BK21 Plus Program, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
2
Department of Chemical and Biomolecular Engineering, BK21 Plus Program, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea; Institute for the BioCentury, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea. Electronic address: kjjeong@kaist.ac.kr.

Abstract

Foot-and-mouth disease (FMD) is caused by the FMD virus (FMDV) and results in severe economic losses in livestock farming. For rapid FMD diagnostic and therapeutic purposes, an effective antibody against FMDV is needed. Here, we developed a high-affinity antibody against FMDV by FACS-based high throughput screening of a random library. With the FITC-conjugated VP1 epitope of FMDV and high-speed FACS sorting, we screened the synthetic antibody (scFv) library in which antibody variants are displayed in the periplasm of Escherichia coli. After three rounds of sorting, we isolated one antibody fragment (#138-scFv) against the VP1 epitope of FMDV. Next, to improve its affinity, a mutation library of #138-scFV was constructed by error-prone PCR and screened by FACS. After three rounds of sorting, we isolated one antibody (AM-32 scFv), which has a higher binding affinity (KD=42.7nM) than that of the original #138-scFv. We also confirmed that it specifically binds to whole inactivated FMDV.

KEYWORDS:

Affinity maturation; Antibody; Escherichia coli; FACS screening; FMDV; Single chain Fv

PMID:
26827774
DOI:
10.1016/j.enzmictec.2015.12.011
[Indexed for MEDLINE]

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