Comprehensive study of domain rearrangements of single-chain bispecific antibodies to determine the best combination of configurations and microbial host cells
| Autor: | Akabane Mihoko, Yuri Kuroki, Hiyamuta Shuichi, Koji Sode, Ryutaro Asano, Watanabe Shunsuke, Izumi Kumagai, Shinzo Mayuzumi, Honma Sachiko |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell Survival medicine.medical_treatment CD3 Recombinant Fusion Proteins Immunology Computational biology medicine.disease_cause Pichia Pichia pastoris Domain (software engineering) law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cancer immunotherapy law Report Cell Line Tumor Antibodies Bispecific medicine Escherichia coli Immunology and Allergy Humans Epidermal growth factor receptor Binding Sites biology Brevibacillus Genetic Variation biology.organism_classification 030104 developmental biology chemistry 030220 oncology & carcinogenesis biology.protein Recombinant DNA Electrophoresis Polyacrylamide Gel Growth inhibition Single-Chain Antibodies |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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