title: High-efficiency generation of monoclonal antibody for Vitreoscilla hemoglobin protein.
abstract: Bacterial hemoglobin from Vitreoscilla (VHb) is recognized as a good fusion protein for the soluble expression of foreign protein. In this study, we generated a monoclonal antibody (MAb) against VHb for its detection. For the rapid screening of MAb, a protein chip technology based on the Alexa-488 (A488) dye labeling method was introduced. In order to fabricate the chip, the VHb protein was chemically coupled to the chip surface and then the culture supernatants of 84 hybridoma cell lines were spotted onto the VHb chip. The bound MAbs were measured by A488- modified anti-mouse IgG. A single spot (MAb A10) exhibited significantly high signal intensity. The immunoblot analysis evidenced that the MAb A10 can detect VHb-fused proteins with high specificity.
authors.affiliation: Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, 900 Gajwa-Dong, Chinju 660-701, Korea.
authors.forename: Eun Mi
authors.initials: EM
authors.lastname: Kim
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kim
authors.forename: Min-Gon
authors.initials: MG
authors.lastname: Kim
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Kim
authors.forename: Hyeon-Su
authors.initials: HS
authors.lastname: Ro
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology* / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000276:immunology*; D005456:Fluorescent Dyes / Q000378:metabolism; D051379:Mice; D046228:Microarray Analysis; D040081:Protein Array Analysis; D013194:Staining and Labeling / Q000379:methods; D054793:Truncated Hemoglobins / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D054793:Truncated Hemoglobins; C082613:hemoglobin protein, Vitreoscilla