公开(公告)号：US20240393297A1

公开(公告)日：2024-11-28

申请号：US18432898

申请日：2024-02-05

Applicant: CureVac Manufacturing GmbH

Inventor： Aniela WOCHNER ,  Isabel REICHERT ,  Kiriaki RAPTOPOULOU

IPC: G01N30/34 ,  B01D15/32 ,  B01D15/36 ,  C12N15/00 ,  C12N15/10 ,  C12Q1/6806 ,  G01N30/00 ,  G01N30/88

Abstract: The present invention relates to the detection and analysis of by-products, such as short RNA transcripts, in a process of RNA in vitro transcription by HPLC. It further relates to the use of this method for the quality control of RNA produced by in vitro transcription or for identifying suitable RNA purification conditions.

公开(公告)号：US20240218412A1

公开(公告)日：2024-07-04

申请号：US18582620

申请日：2024-02-20

Applicant: CureVac Manufacturing GmbH

Inventor： Aniela WOCHNER ,  Tilmann ROOS ,  Thomas KETTERER

IPC: C12P19/34 ,  C12M1/00 ,  C12M1/34 ,  C12M1/36 ,  C12M1/40 ,  C12M3/00 ,  C12Q1/6844

CPC classification number: C12P19/34 ,  C12M21/18 ,  C12M23/44 ,  C12M29/04 ,  C12M29/14 ,  C12M29/18 ,  C12M41/26 ,  C12M41/32 ,  C12M41/48 ,  C12Q1/6844

Abstract: The present invention relates to a method for synthesizing an RNA molecule of a given sequence, comprising the step of determining the fraction (1) for each of the four nucleotides G, A, C and U in said RNA molecule, and the step of synthesizing said RNA molecule by in vitro transcription in a sequence-optimized reaction mix, wherein said sequence-optimized reaction mix comprises the four ribonucleoside triphosphates GTP, ATP, CTP and UTP, wherein the fraction (2) of each of the four ribonucleoside triphosphates in the sequence-optimized reaction mix corresponds to the fraction (1) of the respective nucleotide in said RNA molecule, a buffer, a DNA template, and an RNA polymerase.
Further, the present invention relates to a bioreactor (1) for synthesizing RNA molecules of a given sequence, the bioreactor (1) having a reaction module (2) for carrying out in vitro RNA transcription reactions in a sequence-optimized reaction mix, a capture module (3) for temporarily capturing the transcribed RNA molecules, and a control module (4) for controlling the infeed of components of the sequence-optimized reaction mix into the reaction module (2), wherein the reaction module (2) comprises a filtration membrane (21) for separating nucleotides from the reaction mix, and the control of the infeed of components of the sequence-optimized reaction mix by the control module (4) is based on a measured concentration of separated nucleotides.

公开(公告)号：US20230313268A1

公开(公告)日：2023-10-05

申请号：US18050940

申请日：2022-10-28

Applicant: CureVac Manufacturing GmbH

Inventor： Aniela WOCHNER

IPC: C12Q1/6806 ,  B01D15/16 ,  B01D15/32 ,  C07H21/02 ,  G01N30/88

CPC classification number: C12Q1/6806 ,  B01D15/163 ,  B01D15/325 ,  C07H21/02 ,  G01N30/88 ,  G01N2030/8827

Abstract: The present invention relates to the field of RNA analysis. In particular, the invention concerns the use of a catalytic nucleic acid molecule for the analysis of an RNA molecule. The invention concerns methods for analyzing the 5′ terminal structures of an RNA molecule having a cleavage site for a catalytic nucleic acid molecule. In particular, the invention concerns a method for determining the presence of a cap structure in an RNA molecule having a cleavage site for a catalytic nucleic acid molecule, a method for determining the capping degree of a population of RNA molecules having a cleavage site for a catalytic nucleic acid molecule, a method for determining the orientation of the cap structure in a capped RNA molecule having a cleavage site for a catalytic nucleic acid molecule and a method for determining relative amounts of correctly capped RNA molecules and reverse-capped RNA molecules in a population of RNA molecules, wherein the population comprises correctly capped and/or reverse-capped RNA molecules that have a cleavage site for a catalytic nucleic acid molecule. Moreover, the present invention provides uses of a catalytic nucleic acid molecule.