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-...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-05-01
|
| Series: | Antibodies |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4468/10/2/18 |
| _version_ | 1797534172296773632 |
|---|---|
| author | Stefania C. Carrara David Fiebig Jan P. Bogen Julius Grzeschik Björn Hock Harald Kolmar |
| author_facet | Stefania C. Carrara David Fiebig Jan P. Bogen Julius Grzeschik Björn Hock Harald Kolmar |
| author_sort | Stefania C. Carrara |
| collection | DOAJ |
| 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 (<i>hc</i>) and light chain (<i>lc</i>) genes encoded on separate plasmids. In this study, we examine the production of two FDA-approved antibodies using a bidirectional (BiDi) vector encoding both <i>hc</i> and <i>lc</i> with mirrored promoter and enhancer elements on a single plasmid, by analysing the individual <i>hc</i> and <i>lc</i> 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. |
| first_indexed | 2024-03-10T11:25:50Z |
| format | Article |
| id | doaj.art-618caab871564ae9a80491b80eafe118 |
| institution | Directory Open Access Journal |
| issn | 2073-4468 |
| language | English |
| last_indexed | 2024-03-10T11:25:50Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj.art-618caab871564ae9a80491b80eafe1182023-11-21T19:39:41ZengMDPI AGAntibodies2073-44682021-05-011021810.3390/antib10020018Recombinant Antibody Production Using a Dual-Promoter Single Plasmid SystemStefania C. Carrara0David Fiebig1Jan P. Bogen2Julius Grzeschik3Björn Hock4Harald Kolmar5Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Str. 4, D-64287 Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Str. 4, D-64287 Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Str. 4, D-64287 Darmstadt, GermanyFerring Darmstadt Laboratories, Alarich-Weiss-Str. 4, D-64287 Darmstadt, GermanyFerring International Center S.A, Chemin de la Vergognausaz 50, CH-1162 Saint Prex, SwitzerlandInstitute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Str. 4, D-64287 Darmstadt, GermanyMonoclonal 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 (<i>hc</i>) and light chain (<i>lc</i>) genes encoded on separate plasmids. In this study, we examine the production of two FDA-approved antibodies using a bidirectional (BiDi) vector encoding both <i>hc</i> and <i>lc</i> with mirrored promoter and enhancer elements on a single plasmid, by analysing the individual <i>hc</i> and <i>lc</i> 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.https://www.mdpi.com/2073-4468/10/2/18monoclonal antibodiespromotersbidirectionalantibody productionupstream processing |
| spellingShingle | Stefania C. Carrara David Fiebig Jan P. Bogen Julius Grzeschik Björn Hock Harald Kolmar Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System Antibodies monoclonal antibodies promoters bidirectional antibody production upstream processing |
| 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 | monoclonal antibodies promoters bidirectional antibody production upstream processing |
| url | https://www.mdpi.com/2073-4468/10/2/18 |
| work_keys_str_mv | AT stefaniaccarrara recombinantantibodyproductionusingadualpromotersingleplasmidsystem AT davidfiebig recombinantantibodyproductionusingadualpromotersingleplasmidsystem AT janpbogen recombinantantibodyproductionusingadualpromotersingleplasmidsystem AT juliusgrzeschik recombinantantibodyproductionusingadualpromotersingleplasmidsystem AT bjornhock recombinantantibodyproductionusingadualpromotersingleplasmidsystem AT haraldkolmar recombinantantibodyproductionusingadualpromotersingleplasmidsystem |