Rapid generation of functional human IgG antibodies derived from Fab-on-phage display libraries.
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.
Jostock
Thomas
T
Dyax s.a., Sart Tilman, Liege, Belgium.
Vanhove
Marc
M
Brepoels
Eddy
E
Van Gool
Rein
R
Daukandt
Marc
M
Wehnert
Anita
A
Van Hegelsom
Rob
R
Dransfield
Dan
D
Sexton
Dan
D
Devlin
Mary
M
Ley
Art
A
Hoogenboom
Hennie
H
Müllberg
Jürgen
J
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
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D019151:Peptide Library; D042810:Receptor, TIE-1