title: Rapid generation of functional human IgG antibodies derived from Fab-on-phage display libraries.
abstract: We introduce a procedure for the rapid generation of fully human antibodies derived from "Fab-on-phage" display libraries. The technology is based on the compatibility of display vectors and IgG expression constructs, and allows reformatting of individual Fab clones to IgG, as well as reformatting of antibody repertoires. Examples of batch reformatting of an uncharacterized Fab repertoire and of a pool of Fabs, previously analyzed at the phage level, are presented. The average transient expression levels of the IgG constructs in HEK293T cells are above 10 microg/ml, allowing the use of conditioned media in functional assays without antibody purification. Furthermore, we describe a high-throughput purification method yielding IgG amounts sufficient for initial antibody characterization. Our technology allows the generation and production of antigen-specific complete human antibodies as fast or even faster than raising monoclonal antibodies by conventional hybridoma techniques.
authors.affiliation: Dyax s.a., Sart Tilman, Liege, Belgium.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jostock
authors.forename: Marc
authors.initials: M
authors.lastname: Vanhove
authors.forename: Eddy
authors.initials: E
authors.lastname: Brepoels
authors.forename: Rein
authors.initials: R
authors.lastname: Van Gool
authors.forename: Marc
authors.initials: M
authors.lastname: Daukandt
authors.forename: Anita
authors.initials: A
authors.lastname: Wehnert
authors.forename: Rob
authors.initials: R
authors.lastname: Van Hegelsom
authors.forename: Dan
authors.initials: D
authors.lastname: Dransfield
authors.forename: Dan
authors.initials: D
authors.lastname: Sexton
authors.forename: Mary
authors.initials: M
authors.lastname: Devlin
authors.forename: Art
authors.initials: A
authors.lastname: Ley
authors.forename: Hennie
authors.initials: H
authors.lastname: Hoogenboom
authors.forename: Jürgen
authors.initials: J
authors.lastname: Müllberg
mesh_terms: D000906:Antibodies / Q000235:genetics; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics; D019151:Peptide Library; D042810:Receptor, TIE-1 / Q000276:immunology
chemical_list: D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D019151:Peptide Library; D042810:Receptor, TIE-1