title: Optimized Generation of High-Affinity, High-Specificity Single-Chain Fv Antibodies from Multi-Antigen Immunized Chickens.
abstract: High-affinity, highly specific binding proteins are a key class of molecules used in the development of new affinity chromatography methods. Traditionally, antibody-based methods have relied on the use of immunoglobulins purified from immune animal sera, from egg yolks, or from murine monoclonal hybridoma supernatants. To accelerate and refine the reagent antibody generation process, we have developed optimized methods that allow the rapid assembly of scFv libraries from chickens immunized with pools of immunogens. These methods allow the simplified generation of a single, moderately sized library of single chain Fv (scFv) and the subsequent isolation of diverse, high affinity, and high specificity monoclonals for each individual immunogen, via phage display. Using these methods, antibodies can be derived that exhibit the desired selectivity, including exquisite specificity or cross-reactivity to multiple orthologues of the same protein.
authors.affiliation: Global Biotherapeutic Technologies, Pfizer, Grange Castle Business Park, Clondalkin, Dublin 22, Ireland.
authors.forename: William J J
authors.initials: WJ
authors.lastname: Finlay
authors.affiliation: Pfizer Inc., 610 Main St., Cambridge, MA, 02139, USA.
authors.forename: Laird
authors.initials: L
authors.lastname: Bloom
authors.affiliation: Global Biotherapeutic Technologies, Pfizer, Grange Castle Business Park, Clondalkin, Dublin 22, Ireland.
authors.forename: Sreeja
authors.initials: S
authors.lastname: Varghese
authors.affiliation: Global Biotherapeutic Technologies, Pfizer, Grange Castle Business Park, Clondalkin, Dublin 22, Ireland.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Autin
authors.affiliation: Global Biotherapeutic Technologies, Pfizer, Grange Castle Business Park, Clondalkin, Dublin 22, Ireland. orla.cunningham@pfizer.com.
authors.forename: Orla
authors.initials: O
authors.lastname: Cunningham
mesh_terms: D000818:Animals; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D061505:Cell Surface Display Techniques; D002645:Chickens / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D051379:Mice; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000941:Antigens; D000939:Epitopes; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies
keywords: Affinity; Chicken; Chromatography; Epitope; Monoclonal; Phage display; Recombinant; scFv