Generation of bispecific IgG antibodies by structure-based design of an orthogonal Fab interface.
Robust generation of IgG bispecific antibodies has been a long-standing challenge. Existing methods require extensive engineering of each individual antibody, discovery of common light chains, or complex and laborious biochemical processing. Here we combine computational and rational design approaches with experimental structural validation to generate antibody heavy and light chains with orthogonal Fab interfaces. Parental monoclonal antibodies incorporating these interfaces, when simultaneously co-expressed, assemble into bispecific IgG with improved heavy chain-light chain pairing. Bispecific IgGs generated with this approach exhibit pharmacokinetic and other desirable properties of native IgG, but bind target antigens monovalently. As such, these bispecific reagents may be useful in many biotechnological applications.
1] Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA. [2].
Steven M
SM
Lewis
1] Eli Lilly Biotechnology Center, San Diego, California, USA. [2].
Xiufeng
X
Wu
Eli Lilly Biotechnology Center, San Diego, California, USA.
Anna
A
Pustilnik
Eli Lilly Biotechnology Center, San Diego, California, USA.
Arlene
A
Sereno
Eli Lilly Biotechnology Center, San Diego, California, USA.
Flora
F
Huang
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Heather L
HL
Rick
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Gurkan
G
Guntas
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Andrew
A
Leaver-Fay
Eli Lilly Biotechnology Center, San Diego, California, USA.
Eric M
EM
Smith
Eli Lilly Biotechnology Center, San Diego, California, USA.
Carolyn
C
Ho
Eli Lilly Biotechnology Center, San Diego, California, USA.
Christophe
C
Hansen-Estruch
Eli Lilly Biotechnology Center, San Diego, California, USA.
Aaron K
AK
Chamberlain
Eli Lilly Biotechnology Center, San Diego, California, USA.
Stephanie M
SM
Truhlar
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Elaine M
EM
Conner
Eli Lilly Biotechnology Center, San Diego, California, USA.
Shane
S
Atwell
1] Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA. [2] Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Brian
B
Kuhlman
Eli Lilly Biotechnology Center, San Diego, California, USA.
Stephen J
SJ
Demarest
D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry* / Q000378:metabolism; D001709:Biotechnology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D051379:Mice; D008954:Models, Biological; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*
D018033:Antibodies, Bispecific; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G