| Title: |
Detection of residual T7 RNA polymerase used in mRNA in vitro transcription by Simple Western |
| Authors: |
Shieh, Yvonne; Swartz, Andrew R.; Rustandi, Richard R. |
| Source: |
ELECTROPHORESIS ; volume 45, issue 19-20, page 1834-1839 ; ISSN 0173-0835 1522-2683 |
| Publisher Information: |
Wiley |
| Publication Year: |
2024 |
| Collection: |
Wiley Online Library (Open Access Articles via Crossref) |
| Description: |
Therapeutic messenger RNA (mRNA) has been demonstrated as a scalable and versatile vaccine platform for the rapid development and manufacture of new vaccine candidates. mRNA is synthesized enzymatically through in vitro transcription (IVT) using bacteriophage T7 RNA polymerase (T7 RNAP), a 99 kDa protein with high binding affinity for the promoter sequence and a low error rate. Post‐IVT, mRNA is purified to remove impurities, but if T7 RNAP is insufficiently cleared, undesirable clinical side effects may result. Therefore, it is important to quantitate T7 RNAP concentrations in IVT and process intermediates to understand clearance during downstream purification. A high‐throughput T7 RNAP assay was developed using Simple Western (SW), a capillary immunoassay technology, to quantitate concentrations as low as 5.3 ng/mL with good precision and accuracy. Compared to existing T7 RNAP immunoassays or total protein assays such as bicinchoninic acid assays or Bradford, the SW T7 RNAP assay is specific to T7 RNAP, requires |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1002/elps.202400044 |
| Availability: |
http://dx.doi.org/10.1002/elps.202400044 |
| Rights: |
http://creativecommons.org/licenses/by/4.0/ |
| Accession Number: |
edsbas.7FEB469E |
| Database: |
BASE |