Immobilized poly(n)polymerase
AssigneeCureVac AG
InventorsTilmann Roos, Benyamin Yazdan Panah, Markus Conzelmann, Veronika Wagner
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Abstract
The present invention relates to an immobilized poly(N)polymerase (PNP), methods of producing said PNP and uses thereof. Further disclosed is an enzyme reactor and kit comprising the PNP for producing polynucleotidylated ribonucleic acid poly(N)RNA)molecules which are useful in gene therapy, immunotherapy, protein replacement therapy and/or vaccination.
Claims17
A method of producing polyadenylated ribonucleic acid (poly(A)RNA) molecules comprising contacting an RNA molecule with a poly(A)polymerase covalently immobilized onto a solid support, said poly(A)polymerase comprising at least 90% sequence identity to SEQ ID NO:
- 113, wherein the poly(A)polymerase comprises at least one newly introduced cysteine residue compared to a native poly(A)polymerase, wherein the at least one newly introduced cysteine residue is located at a terminus of the protein, wherein the poly(A)polymerase is immobilized via a thiol group of the at least one newly introduced cysteine residue, and wherein the at least one cysteine residue is attached to the poly(A)polymerase via a peptide linker element comprising the sequence of SEQ ID NO:
- 162 (GlyGlyGlyGly), and,
- wherein at least 80% of the produced poly(A)RNA has a poly(A) length of at least 100 nucleotides.
Dependencies are inferred from the claim text.