Humanization of antibodies using heavy chain complementarity-determining region 3 grafting coupled with in vitro somatic hypermutation.
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.
Anaptysbio Inc., San Diego, California 92131. Electronic address: pbowers@anaptysbio.com.
Peter M
PM
Bowers
Anaptysbio Inc., San Diego, California 92131.
Tamlyn Y
TY
Neben
Anaptysbio Inc., San Diego, California 92131.
Geoffery L
GL
Tomlinson
Anaptysbio Inc., San Diego, California 92131.
Jennifer L
JL
Dalton
Anaptysbio Inc., San Diego, California 92131.
Larry
L
Altobell
Anaptysbio Inc., San Diego, California 92131.
Xue
X
Zhang
Anaptysbio Inc., San Diego, California 92131.
John L
JL
Macomber
Anaptysbio Inc., San Diego, California 92131.
Betty F
BF
Wu
Anaptysbio Inc., San Diego, California 92131.
Rachelle M
RM
Toobian
Anaptysbio Inc., San Diego, California 92131.
Audrey D
AD
McConnell
Anaptysbio Inc., San Diego, California 92131.
Petra
P
Verdino
Anaptysbio Inc., San Diego, California 92131.
Betty
B
Chau
Anaptysbio Inc., San Diego, California 92131.
Robert A
RA
Horlick
Anaptysbio Inc., San Diego, California 92131.
David J
DJ
King
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000941:Antigens / Q000737:chemistry; D001483:Base Sequence; D001667:Binding, Competitive; D002469:Cell Separation; D003062:Codon; D022801:Complementarity Determining Regions / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D015202:Protein Engineering / Q000379:methods; D015398:Signal Transduction
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D003062:Codon; D022801:Complementarity Determining Regions; D016207:Cytokines