title: Development of high-affinity single chain Fv against foot-and-mouth disease virus.
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.
authors.affiliation: Department of Chemical and Biomolecular Engineering, BK21 Plus Program, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
authors.forename: Joon-Goo
authors.initials: JG
authors.lastname: Jung
authors.affiliation: Department of Chemical and Biomolecular Engineering, BK21 Plus Program, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
authors.forename: Gu Min
authors.initials: GM
authors.lastname: Jeong
authors.affiliation: Department of Chemical and Biomolecular Engineering, BK21 Plus Program, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.
authors.forename: Sung Sun
authors.initials: SS
authors.lastname: Yim
authors.affiliation: 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.
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000915:Antibody Affinity; D036022:Capsid Proteins / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D005536:Foot-and-Mouth Disease / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*; D020349:Surface Plasmon Resonance
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C442501:VP1 protein, Foot-and-mouth disease virus
keywords: Affinity maturation; Antibody; Escherichia coli; FACS screening; FMDV; Single chain Fv