METHOD FOR PRODUCING RNA

摘要

The present invention relates to a method for producing RNA. In particular, the present invention relates to a method for producing RNA, which is scalable and provides RNA at a high purity. The present invention provides a method for producing RNA under GMP and/or cGMP-compliant conditions. The invention further provides specific processes for use as a quality control in the manufacturing of a template DNA and/or in a method for producing RNA, in particular by in vitro transcription.

主权项

1. A method for producing purified RNA comprising the following steps:
a) providing a template DNA comprising a nucleic acid sequence encoding an RNA sequence, wherein the following steps are used to control the quality of the template DNA provided in step a):
I) determining the concentration of the template DNA in a sample;II) determining the integrity of the template DNA;III) determining the identity of the template DNA; andIV) determining the purity of the template DNA; and b) in vitro transcription of the template DNA in order to obtain a composition comprising the RNA; and c) purifying the RNA by at least one purification step selected from the group consisting of a precipitation step and a chromatographic step to obtain purified RNA, wherein the following steps are used to assess the quality of the RNA obtained in step b or the purified RNA obtained in step c:
i) determining the concentration of the RNA or the purified RNA in a sample;ii) determining the integrity of the RNA or the purified RNA;iii) determining the identity of the RNA or the purified RNA;iv) determining the purity of the RNA or the purified RNA;v) determining the pH of a sample comprising the RNA or the purified RNA;vi) determining the osmolality of a sample comprising the RNA or the purified RNA;vii) determining the presence and/or the amount of the template DNA in a sample comprising the RNA or the purified RNA; andviii) determining the presence and/or the amount of an organic solvent in a sample comprising the RNA or the purified RNA.