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
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2021 |
| Autor(en): | Carrara, Stefania C. ; Fiebig, David ; Bogen, Jan P. ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald |
| Art des Eintrags: | Bibliographie |
| Titel: | Recombinant antibody production using a dual-promoter single plasmid system |
| Sprache: | Englisch |
| Publikationsjahr: | 2021 |
| Ort: | Basel |
| Verlag: | MDPI |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Antibodies |
| Jahrgang/Volume einer Zeitschrift: | 10 |
| (Heft-)Nummer: | 2 |
| Kollation: | 12 Seiten |
| DOI: | 10.3390/antib10020018 |
| Zugehörige Links: | |
| Kurzbeschreibung (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. |
| Freie Schlagworte: | monoclonal antibodies, promoters, bidirectional, antibody production, upstream processing |
| Zusätzliche Informationen: | This article belongs to the Special Issue Design, Production and Characterization of Peptide Antibodies |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie |
| Hinterlegungsdatum: | 15 Jan 2024 07:47 |
| Letzte Änderung: | 16 Jan 2024 07:54 |
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| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
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Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System. (deposited 12 Jan 2024 15:01)
- Recombinant antibody production using a dual-promoter single plasmid system. (deposited 15 Jan 2024 07:47) [Gegenwärtig angezeigt]
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