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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
Artikel, Bibliographie

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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
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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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