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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-...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-8161450 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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