Tethered-variable CL bispecific IgG: an antibody platform for rapid bispecific antibody screening

Autor: James A. Ernst, Christoph Spiess, Angie Yee, Hok Seon Kim, Jo-Anne Hongo, Ingrid Kim, Raymond K. Tong, Diana Ronai Dunshee, Farzam Farahi, Junichiro Sonoda
Rok vydání: 2017
Předmět:
0301 basic medicine
Bispecific antibody
Gene Expression
Protein Engineering
Biochemistry
Mice
FGF21
0302 clinical medicine
Antibodies
Bispecific
Cloning
Molecular
Mice
Inbred BALB C
biology
Drug discovery
Chemistry
Antibodies
Monoclonal
Recombinant Proteins
bispecific antibody
030220 oncology & carcinogenesis
Original Article
Antibody
Biotechnology
medicine.drug_class
High-throughput screening
Genetic Vectors
Bioengineering
CHO Cells
Computational biology
therapeutic mAb
Immunoglobulin light chain
Monoclonal antibody
03 medical and health sciences
Cricetulus
biepitopic antibody
medicine
Animals
Humans
Receptor
Fibroblast Growth Factor
Type 1
Klotho Proteins
Molecular Biology
Membrane Proteins
High-Throughput Screening Assays
Fibroblast Growth Factors
FGFR1
HEK293 Cells
030104 developmental biology
Immunoglobulin G
biology.protein
KLB
Merge (version control)
high throughput screening
Zdroj: Protein Engineering, Design and Selection
ISSN: 1741-0134
1741-0126
Popis: Bispecific antibodies offer a clinically validated platform for drug discovery. In generating functionally active bispecific antibodies, it is necessary to identify a unique parental antibody pair to merge into a single molecule. However, technologies that allow high-throughput production of bispecific immunoglobulin Gs (BsIgGs) for screening purposes are limited. Here, we describe a novel bispecific antibody format termed tethered-variable CLBsIgG (tcBsIgG) that allows robust production of intact BsIgG in a single cell line, concurrently ensuring cognate light chain pairing and preserving key antibody structural and functional properties. This technology is broadly applicable in the generation of BsIgG from a variety of antibody isotypes, including human BsIgG1, BsIgG2 and BsIgG4. The practicality of the tcBsIgG platform is demonstrated by screening BsIgGs generated from FGF21-mimetic anti-Klotho-β agonistic antibodies in a combinatorial manner. This screen identified multiple biepitopic combinations with enhanced agonistic activity relative to the parental monoclonal antibodies, thereby demonstrating that biepitopic antibodies can acquire enhanced functionality compared to monospecific parental antibodies. By design, the tcBsIgG format is amenable to high-throughput production of large panels of bispecific antibodies and thus can facilitate the identification of rare BsIgG combinations to enable the discovery of molecules with improved biological function.
Databáze: OpenAIRE
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