SO3 chromatography for use in a method for virus purification
Inventors
Wolschek, Markus • Reiter, Manfred • Strancar, Ales • SBAIZERO, Mojca Tajnik
Assignees
Blue Sky Vaccines GmbH • Bluesky Immunotherapies GmbH • Sartorius Bia Separations doo
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11339377-B2
Publication Date
2022-05-24
Expiration Date
Abstract
The present invention refers to a process for purifying virus particles from cell culture, comprising the steps of subjecting the cell culture to centrifugation to get a supernatant fraction of virus particles, incubating said fraction with a nuclease, diluting the fraction with low conductivity buffer, subjecting said diluted fraction to a SO3 chromatography step, performing a washing step with low conductivity buffer, and eluting virus particles.
Core Innovation
The invention relates to a process for purifying virus particles from cell culture. The process comprises subjecting the cell culture to centrifugation to obtain a supernatant fraction of virus particles, incubating the supernatant fraction with a nuclease, and diluting the nuclease-treated supernatant fraction with a low conductivity dilution buffer to obtain a diluted fraction.
The diluted fraction is subjected to a SO3 chromatography step, followed by a washing step with a low conductivity wash buffer having a conductivity of 12 mS/cm or less before the method elutes the virus particles. The SO3 chromatography is described for purifying influenza virus from cell culture while maintaining infectivity/immunogenicity.
The disclosed approach centers on controlling conductivity around the SO3 chromatography step by using low conductivity dilution and low conductivity wash conditions, and by controlling nuclease treatment and nuclease carryover. The document further describes buffering and formulation components in the low conductivity buffers and provides example results showing improved SO3 binding/dynamic binding capacity when harvest conductivity is reduced and strong depletion of host cell protein and residual DNA.
Claims Coverage
The document includes one independent claim, defining a sequential purification process with SO3 chromatography and low-conductivity dilution and wash conditions, followed by elution. The independent claim is built from main inventive features that are refined in dependent claims by specifying quantitative removal constraints and operational constraints, including conductivity and centrifugation thresholds.
Sequential virus purification with nuclease-treated, low-conductivity SO3 chromatography
A process for purifying virus particles from cell culture comprising, in the following order, subjecting the cell culture to centrifugation to obtain a supernatant fraction of virus particles; incubating the supernatant fraction with a nuclease; diluting the supernatant fraction with a low conductivity dilution buffer to obtain a diluted fraction; subjecting the diluted fraction to a SO3 chromatography step; performing a washing step with low conductivity wash buffer having a conductivity of 12 mS/cm or less; and eluting virus particles.
Across the independent claim and its refinements, the core coverage is a nuclease incubation followed by dilution with a low conductivity buffer and SO3 chromatography, combined with a low conductivity wash of 12 mS/cm or less before elution. Dependent claims further narrow the process by adding quantifiable host-cell protein and DNA removal targets, specifying centrifugation speed thresholds, and defining elution modes and wash components such as EDTA.
Stated Advantages
Improved SO3 binding/dynamic binding capacity when harvest conductivity is reduced.
Strong depletion of host cell protein.
Residual DNA is depleted.
Maintaining infectivity/immunogenicity.
Reduction of nuclease carryover using EDTA in wash buffers.
Documented Applications
Purification of influenza virus from cell culture.
Interested in licensing this patent?