Removal of DNA fragments in mRNA production process
US10077439B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Mar 13, 2014 |
| Grant date | Sep 18, 2018 |
| Priority date | — |
| Expiry date | Mar 13, 2034 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6806
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention describes methods of removing DNA from an RNA transcript during the mRNA production process. The method embodies procedures for obtaining an in vitro transcription product, and removing any DNA from the product. The DNA can be removed by adding either free DNase or a resin containing immobilized DNase to the product, and recovering the RNA transcript. Alternatively, the DNA template used in the in vitro transcription reaction is labeled. After transcription, the product is applied to a resin that is configured to bind the label, and the RNA transcript is recovered. To detect whether any residual impurities are left in the RNA transcript product, the product is subjected to nuclease digestion and subsequently to liquid chromatography-tandem mass spectrometry analysis to quantitate any residual DNA. The present invention demonstrates efficient and effective methods of isolating an RNA transcript from an in vitro transcription product.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.