Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells

BACKGROUND: Two-armed FabscFv-Fc is a favoured bispecific antibody (BsAb) format due to its advantages of the conventional IgG structure. Production of FabscFv-Fc requires expression of three polypeptide chains, one light chain (LC), one heavy chain (HC) and a scFv fused to the Fc (scFvFc) at optima...

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Detalles Bibliográficos
Autores principales: Ong, Han Kee, Nguyen, Ngan T B, Bi, Jiawu, Yang, Yuansheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Original research article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9743168/
https://www.ncbi.nlm.nih.gov/pubmed/36518226
http://dx.doi.org/10.1093/abt/tbac025
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author Ong, Han Kee
Nguyen, Ngan T B
Bi, Jiawu
Yang, Yuansheng
author_facet Ong, Han Kee
Nguyen, Ngan T B
Bi, Jiawu
Yang, Yuansheng
author_sort Ong, Han Kee
collection PubMed
description BACKGROUND: Two-armed FabscFv-Fc is a favoured bispecific antibody (BsAb) format due to its advantages of the conventional IgG structure. Production of FabscFv-Fc requires expression of three polypeptide chains, one light chain (LC), one heavy chain (HC) and a scFv fused to the Fc (scFvFc) at optimal ratios. METHODS: We designed a set of internal ribosome entry site (IRES)-mediated multi-cistronic vectors tailoring to various expression ratios of the three polypeptides to study how the chain ratios affect the FabscFv-Fc production. RESULTS: Expression of HC and scFvFc chains at 1:1 ratio and excess LC gave the highest yield of correctly assembled product. Compared to the use of IRES and multiple promoters, using 2A peptides for co-expression of the three polypeptides gave the highest titre and correctly assembled product. CONCLUSION: The results obtained in this work provide insights to the impacts of hetero-chain ratios on the BsAb production.
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spelling pubmed-97431682022-12-13 Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells Ong, Han Kee Nguyen, Ngan T B Bi, Jiawu Yang, Yuansheng Antib Ther Original research article BACKGROUND: Two-armed FabscFv-Fc is a favoured bispecific antibody (BsAb) format due to its advantages of the conventional IgG structure. Production of FabscFv-Fc requires expression of three polypeptide chains, one light chain (LC), one heavy chain (HC) and a scFv fused to the Fc (scFvFc) at optimal ratios. METHODS: We designed a set of internal ribosome entry site (IRES)-mediated multi-cistronic vectors tailoring to various expression ratios of the three polypeptides to study how the chain ratios affect the FabscFv-Fc production. RESULTS: Expression of HC and scFvFc chains at 1:1 ratio and excess LC gave the highest yield of correctly assembled product. Compared to the use of IRES and multiple promoters, using 2A peptides for co-expression of the three polypeptides gave the highest titre and correctly assembled product. CONCLUSION: The results obtained in this work provide insights to the impacts of hetero-chain ratios on the BsAb production. Oxford University Press 2022-10-13 /pmc/articles/PMC9743168/ /pubmed/36518226 http://dx.doi.org/10.1093/abt/tbac025 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Antibody Therapeutics. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original research article
Ong, Han Kee
Nguyen, Ngan T B
Bi, Jiawu
Yang, Yuansheng
Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title_full Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title_fullStr Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title_full_unstemmed Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title_short Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells
title_sort vector design for enhancing expression level and assembly of knob-into-hole based fabscfv-fc bispecific antibodies in cho cells
topic Original research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9743168/
https://www.ncbi.nlm.nih.gov/pubmed/36518226
http://dx.doi.org/10.1093/abt/tbac025
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