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Recombinant antibody production using a dual-promoter single plasmid system

Carrara, Stefania C. ; Fiebig, David ; Bogen, Jan P. ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald (2021)
Recombinant antibody production using a dual-promoter single plasmid system.
In: Antibodies, 10 (2)
doi: 10.3390/antib10020018
Article, Bibliographie

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Abstract

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.

Item Type: Article
Erschienen: 2021
Creators: Carrara, Stefania C. ; Fiebig, David ; Bogen, Jan P. ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald
Type of entry: Bibliographie
Title: Recombinant antibody production using a dual-promoter single plasmid system
Language: English
Date: 2021
Place of Publication: Basel
Publisher: MDPI
Journal or Publication Title: Antibodies
Volume of the journal: 10
Issue Number: 2
Collation: 12 Seiten
DOI: 10.3390/antib10020018
Corresponding Links:
Abstract:

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.

Uncontrolled Keywords: monoclonal antibodies, promoters, bidirectional, antibody production, upstream processing
Additional Information:

This article belongs to the Special Issue Design, Production and Characterization of Peptide Antibodies

Classification DDC: 500 Science and mathematics > 540 Chemistry
500 Science and mathematics > 570 Life sciences, biology
Divisions: 07 Department of Chemistry
07 Department of Chemistry > Clemens-Schöpf-Institut > Fachgebiet Biochemie
Date Deposited: 15 Jan 2024 07:47
Last Modified: 16 Jan 2024 07:54
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