title: Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS).
abstract: Antibodies and their derivatives are the most important agents in therapeutics and diagnostics. Even after the significant progress in the technology for antibody screening from huge libraries, it takes a long time to isolate an antibody, which prevents a prompt action against the spread of a disease. Here, we report a new strategy for isolating desired antibodies from a combinatorial library in one day by repeated fluorescence-activated cell sorting (FACS). First, we constructed a library of synthetic human antibody in which single-chain variable fragment (scFv) was expressed in the periplasm of Escherichia coli. After labeling the cells with fluorescent antigen probes, the highly fluorescent cells were sorted by using a high-speed cell sorter, and these cells were reused without regeneration in the next round of sorting. After repeating this sorting, the positive clones were completely enriched in several hours. Thus, we screened the library against three viral antigens, including the H1N1 influenza virus, Hepatitis B virus, and Foot-and-mouth disease virus. Finally, the potential antibody candidates, which show K(D) values between 10 and 100 nM against the target antigens, could be successfully isolated even though the library was relatively small (∼ 10(6)). These results show that repeated FACS screening without regeneration of the sorted cells can be a powerful method when a rapid response to a spreading disease is required.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea.
authors.forename: Sung Sun
authors.initials: SS
authors.lastname: Yim
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea.
authors.forename: Hyun Bae
authors.initials: HB
authors.lastname: Bang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea.
authors.forename: Young Hwan
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea.
authors.forename: Yong Jae
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea.
authors.forename: Gu Min
authors.initials: GM
authors.lastname: Jeong
authors.affiliation: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) (BK21 plus program), Daejeon, Republic of Korea; Institute for the BioCentury, KAIST, Daejeon, Republic of Korea.
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D005536:Foot-and-Mouth Disease / Q000276:immunology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D006509:Hepatitis B / Q000276:immunology; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology* / Q000302:isolation & purification*
chemical_list: D057127:Single-Chain Antibodies