title: Towards proteome scale antibody selections using phage display.
abstract: In vitro antibody generation by panning a large universal gene library with phage display was employed to generate antibodies to more than 60 different antigens. Of particular interest was a comparison of pannings on 20 different SH2 domains provided by the Structural Genomics Consortium (SGC). Streamlined methods for high throughput antibody generation developed within the 'Antibody Factory' of the German National Genome Research Network (NGFN) were demonstrated to minimise effort and provide a reliable and robust source for antibodies. For the SH2 domains, in two successive series of selections, 2668 clones were analysed, resulting in 347 primary hits in ELISA. Half of these hits were further analysed, and more than 90 different scFv antibodies to all antigens were identified. The validation of selected antibodies by cross-reactivity ELISA, western blot and on protein microarrays demonstrated the versatility of the in vitro antibody selection pipeline to generate a renewable resource of highly specific monoclonal binders in proteome scale numbers with substantially reduced effort and time.
authors.affiliation: Technische Universität Braunschweig, Institute of Biochemistry and Biotechnology, Braunschweig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Mersmann
authors.forename: Doris
authors.initials: D
authors.lastname: Meier
authors.forename: Jana
authors.initials: J
authors.lastname: Mersmann
authors.forename: Saskia
authors.initials: S
authors.lastname: Helmsing
authors.forename: Peter
authors.initials: P
authors.lastname: Nilsson
authors.forename: Susanne
authors.initials: S
authors.lastname: Gräslund
authors.forename: Karen
authors.initials: K
authors.lastname: Colwill
authors.forename: Michael
authors.initials: M
authors.lastname: Hust
authors.forename: Stefan
authors.initials: S
authors.lastname: Dübel
mesh_terms: D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019151:Peptide Library; D020543:Proteome / Q000276:immunology* / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D019151:Peptide Library; D020543:Proteome