title: Surrogate antigens as targets for proteome-wide binder selection.
abstract: In the past decade, many initiatives were taken for the development of antibodies for proteome-wide studies, as well as characterisation and validation of clinically relevant disease biomarkers. Phage display offers many advantages compared to antibody generation by immunisation because it is an unlimited resource of affinity reagents without batch-to-batch variation and is also amendable for high throughput in contrast to conventional hybridoma technology. One of the major bottlenecks to proteome-wide binder selection is the limited supply of suitable target antigens representative of the human proteome. Here, we provide proof of principle of using easily accessible, cancer-associated protein epitope signature tags (PrESTs), routinely generated within the Human Protein Atlas project, as surrogate antigens for full-length proteins in phage selections for the retrieval of target-specific binders. These binders were subsequently tested in western blot, immunohistochemistry and protein microarray application to demonstrate their functionality.
authors.affiliation: Department of Immunotechnology, Lund University, SE-221 84 Lund, Sweden.
authors.forename: Elin
authors.initials: E
authors.lastname: Gustavsson
authors.forename: Sara
authors.initials: S
authors.lastname: Ek
authors.forename: Johanna
authors.initials: J
authors.lastname: Steen
authors.forename: Malin
authors.initials: M
authors.lastname: Kristensson
authors.forename: Cajsa
authors.initials: C
authors.lastname: Älgenäs
authors.forename: Mathias
authors.initials: M
authors.lastname: Uhlén
authors.forename: Christer
authors.initials: C
authors.lastname: Wingren
authors.forename: Jenny
authors.initials: J
authors.lastname: Ottosson
authors.forename: Sophia
authors.initials: S
authors.lastname: Hober
authors.forename: Carl A K
authors.initials: CA
authors.lastname: Borrebaeck
mesh_terms: D000941:Antigens / Q000276:immunology*; D001665:Binding Sites; D015415:Biomarkers; D007118:Immunoassay / Q000379:methods*; D011485:Protein Binding; D020543:Proteome / Q000276:immunology*
chemical_list: D000941:Antigens; D015415:Biomarkers; D020543:Proteome