Beyond CDR-grafting: Structure-guided humanization of framework and CDR regions of an anti-myostatin antibody.
Antibodies are an important class of biotherapeutics that offer specificity to their antigen, long half-life, effector function interaction and good manufacturability. The immunogenicity of non-human-derived antibodies, which can be a major limitation to development, has been partially overcome by humanization through complementarity-determining region (CDR) grafting onto human acceptor frameworks. The retention of foreign content in the CDR regions, however, is still a potential immunogenic liability. Here, we describe the humanization of an anti-myostatin antibody utilizing a 2-step process of traditional CDR-grafting onto a human acceptor framework, followed by a structure-guided approach to further reduce the murine content of CDR-grafted antibodies. To accomplish this, we solved the co-crystal structures of myostatin with the chimeric (Protein Databank (PDB) id 5F3B) and CDR-grafted anti-myostatin antibody (PDB id 5F3H), allowing us to computationally predict the structurally important CDR residues as well as those making significant contacts with the antigen. Structure-based rational design enabled further germlining of the CDR-grafted antibody, reducing the murine content of the antibody without affecting antigen binding. The overall "humanness" was increased for both the light and heavy chain variable regions.
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
James R
JR
Apgar
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Michelle
M
Mader
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Rita
R
Agostinelli
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Susan
S
Benard
b Rare Disease Research Unit, Pfizer Inc. , Cambridge , MA , USA.
Peter
P
Bialek
b Rare Disease Research Unit, Pfizer Inc. , Cambridge , MA , USA.
Mark
M
Johnson
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Yijie
Y
Gao
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Mark
M
Krebs
b Rare Disease Research Unit, Pfizer Inc. , Cambridge , MA , USA.
Jane
J
Owens
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Kevin
K
Parris
b Rare Disease Research Unit, Pfizer Inc. , Cambridge , MA , USA.
Michael
M
St Andre
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Kris
K
Svenson
b Rare Disease Research Unit, Pfizer Inc. , Cambridge , MA , USA.
Carl
C
Morris
a Biomedicine Design, Pfizer Inc. , Cambridge , MA , USA.
Lioudmila
L
Tchistiakova
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D022801:Complementarity Determining Regions / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D055435:Myostatin / Q000276:immunology*; D015202:Protein Engineering / Q000379:methods*
D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; C525911:MSTN protein, human; D055435:Myostatin
Antibody; X-ray crystallography; antibody engineering; complementarity-determining region; humanization; immunogenicity; in silico design; myostatin