US 10,954,492 B2 · Processes for production and purification of nucleic acid-containing compositions

Abstract

Described are improved processes for production and purification of nucleic acid-containing compositions, such as non-naturally occurring viruses, for example, recombinant polioviruses that can be employed as oncolytic agents. Some of the described improved processes relate to improved processes for producing viral DNA template. Also described are improved processes for chromatography purification of nucleic acid-containing compositions, in which the nucleic acid is quantified in chromatography fractions using a rapid detection method of the one or more nucleic acid sequences of the nucleic acid-containing composition, such as detection by real time RT-qPCR. In addition, improved processes for production and purification of oncolytic poliovirus, such as PVS-RIPO, are described. Compositions generated using these methods are also provided.

Claims21

Dependent claims

A purification process for obtaining a composition comprising a live non-naturally occurring poliovirus, comprising:

separating an aqueous fluid comprising the live non-naturally occurring poliovirus on a size separation chromatography column;
measuring by quantitative polymerase chain reaction (qPCR) one or more nucleic acid sequences found in the live non-naturally occurring poliovirus in at least one fraction of the eluate from said size separation column;
collecting at least one positive fraction of the eluate from said size separation column containing the one or more nucleic acid sequences found in the live non-naturally occurring poliovirus;
pooling the at least one positive fraction;
separating the pooled at least one positive fraction on an anion exchange chromatography column;
and
collecting in at least one positive fraction of the flow-through eluate from said anion exchange chromatography column containing the live non-naturally occurring poliovirus.

4 more elements

Dependencies are inferred from the claim text.

Processes for production and purification of nucleic acid-containing compositions

Life of this patent

Processes for production and purification of nucleic acid-containing compositions

Life of this patent

Abstract

Claims21