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Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System

Monoclonal antibodies (mAbs) have demonstrated tremendous effects on the treatment of various disease indications and remain the fastest growing class of therapeutics. Production of recombinant antibodies is performed using mammalian expression systems to facilitate native antibody folding and post-...

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Autores principales: Carrara, Stefania C., Fiebig, David, Bogen, Jan P., Grzeschik, Julius, Hock, Björn, Kolmar, Harald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161450/
https://www.ncbi.nlm.nih.gov/pubmed/34068440
http://dx.doi.org/10.3390/antib10020018
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author Carrara, Stefania C.
Fiebig, David
Bogen, Jan P.
Grzeschik, Julius
Hock, Björn
Kolmar, Harald
author_facet Carrara, Stefania C.
Fiebig, David
Bogen, Jan P.
Grzeschik, Julius
Hock, Björn
Kolmar, Harald
author_sort Carrara, Stefania C.
collection PubMed
description Monoclonal antibodies (mAbs) have demonstrated tremendous effects on the treatment of various disease indications and remain the fastest growing class of therapeutics. Production of recombinant antibodies is performed using mammalian expression systems to facilitate native antibody folding and post-translational modifications. Generally, mAb expression systems utilize co-transfection of heavy chain (hc) and light chain (lc) genes encoded on separate plasmids. In this study, we examine the production of two FDA-approved antibodies using a bidirectional (BiDi) vector encoding both hc and lc with mirrored promoter and enhancer elements on a single plasmid, by analysing the individual hc and lc mRNA expression levels and subsequent quantification of fully-folded IgGs on the protein level. From the assessment of different promoter combinations, we have developed a generic expression vector comprised of mirrored enhanced CMV (eCMV) promoters showing comparable mAb yields to a two-plasmid reference. This study paves the way to facilitate small-scale mAb production by transient cell transfection with a single vector in a cost- and time-efficient manner.
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spelling pubmed-81614502021-05-29 Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System Carrara, Stefania C. Fiebig, David Bogen, Jan P. Grzeschik, Julius Hock, Björn Kolmar, Harald Antibodies (Basel) Article Monoclonal antibodies (mAbs) have demonstrated tremendous effects on the treatment of various disease indications and remain the fastest growing class of therapeutics. Production of recombinant antibodies is performed using mammalian expression systems to facilitate native antibody folding and post-translational modifications. Generally, mAb expression systems utilize co-transfection of heavy chain (hc) and light chain (lc) genes encoded on separate plasmids. In this study, we examine the production of two FDA-approved antibodies using a bidirectional (BiDi) vector encoding both hc and lc with mirrored promoter and enhancer elements on a single plasmid, by analysing the individual hc and lc mRNA expression levels and subsequent quantification of fully-folded IgGs on the protein level. From the assessment of different promoter combinations, we have developed a generic expression vector comprised of mirrored enhanced CMV (eCMV) promoters showing comparable mAb yields to a two-plasmid reference. This study paves the way to facilitate small-scale mAb production by transient cell transfection with a single vector in a cost- and time-efficient manner. MDPI 2021-05-13 /pmc/articles/PMC8161450/ /pubmed/34068440 http://dx.doi.org/10.3390/antib10020018 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Carrara, Stefania C.
Fiebig, David
Bogen, Jan P.
Grzeschik, Julius
Hock, Björn
Kolmar, Harald
Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title_full Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title_fullStr Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title_full_unstemmed Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title_short Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
title_sort recombinant antibody production using a dual-promoter single plasmid system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161450/
https://www.ncbi.nlm.nih.gov/pubmed/34068440
http://dx.doi.org/10.3390/antib10020018
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