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Comprehensive study of domain rearrangements of single-chain bispecific antibodies to determine the best combination of configurations and microbial host cells.
- Source :
-
MAbs [MAbs] 2018 Aug/Sep; Vol. 10 (6), pp. 854-863. Date of Electronic Publication: 2018 Jul 09. - Publication Year :
- 2018
-
Abstract
- Small bispecific antibodies (bsAbs) are important therapeutic molecules and represent the first bsAb format approved by the United States Food and Drug Administration. Diabody (Db), a small bsAb format, has four possible domain orders; we previously reported the differences in the expression levels and cancer growth inhibition effects upon rearranging the domain order of this format. However, there have been no comprehensive reports on domain rearrangements of bispecific single-chain Db (scDb) and tandem single-chain Fv (taFv), which are widely used bsAb formats. In this study, we designed all possible domain orders for scDb and taFv (each with eight variants) with identical Fv pairs and individually expressed all 16 variants using Escherichia coli, Pichia pastoris, and Brevibacillus choshinensis. Comprehensive investigations showed that the intrinsic functions of the variants were similar to each other, regardless of the expression host system, but expression levels varied depending on the format as well as on the host cell. Among the 16 variants, we found a promising candidate that exhibited high activity and productivity. Furthermore, we determined that B. choshinensis is an attractive expression host because of its secretory production of recombinant proteins.
- Subjects :
- Antibodies, Bispecific genetics
Binding Sites genetics
Brevibacillus genetics
Cell Line, Tumor
Cell Survival immunology
Electrophoresis, Polyacrylamide Gel
Escherichia coli genetics
Genetic Variation
Humans
Pichia genetics
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Single-Chain Antibodies genetics
Antibodies, Bispecific immunology
Binding Sites immunology
Recombinant Fusion Proteins immunology
Single-Chain Antibodies immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1942-0870
- Volume :
- 10
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- MAbs
- Publication Type :
- Academic Journal
- Accession number :
- 29985753
- Full Text :
- https://doi.org/10.1080/19420862.2018.1476815