Humanization of Antibodies Using Heavy Chain Complementarity-determining Region 3 Grafting Coupled with in Vitro Somatic Hypermutation

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2013 Jan 29

PMID 23355464

Citations 11

Authors

Peter M Bowers

Tamlyn Y Neben

Geoffery L Tomlinson

Jennifer L Dalton

Larry Altobell

Xue Zhang

John L Macomber

Betty F Wu

Rachelle M Toobian

Audrey D McConnell

Petra Verdino

Betty Chau

Robert A Horlick

David J King

Affiliations

Soon will be listed here.

Abstract

A method for simultaneous humanization and affinity maturation of monoclonal antibodies has been developed using heavy chain complementarity-determining region (CDR) 3 grafting combined with somatic hypermutation in vitro. To minimize the amount of murine antibody-derived antibody sequence used during humanization, only the CDR3 region from a murine antibody that recognizes the cytokine hβNGF was grafted into a nonhomologous human germ line V region. The resulting CDR3-grafted HC was paired with a CDR-grafted light chain, displayed on the surface of HEK293 cells, and matured using in vitro somatic hypermutation. A high affinity humanized antibody was derived that was considerably more potent than the parental antibody, possessed a low pm dissociation constant, and demonstrated potent inhibition of hβNGF activity in vitro. The resulting antibody contained half the heavy chain murine donor sequence compared with the same antibody humanized using traditional methods.

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