Virus purification
Inventors
Barbero Calzado, Jana • Nebenführ, Mario • Schlegl, Robert • Weber, Michael • HEINDL-WRUSS, Jürgen
Assignees
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Publication Number
US-12214033-B2
Publication Date
2025-02-04
Expiration Date
Abstract
Described herein are processes for purifying infectious virus particles and uses of protamine in such processes.
Core Innovation
The invention relates to separating and purifying infectious virus particles from non-infectious virus particles and impurities by using protamine. The approach includes precipitating non-infectious virus particles with protamine to obtain a virus preparation enriched in infectious virus particles, with the disclosed methods targeting virus particles selected from specified virus families and expressly excluding Zika virus particles.
In one aspect, a crude harvest comprising infectious virus particles, non-infectious virus particles, and impurities generated from growing virus particles on a cell substrate is contacted with an agent comprising protamine. The invention provides an enrichment of infectious virus particles in the resulting virus preparation relative to total virus particles in the crude harvest, within the range of at least 50% to 95%.
In another aspect, the invention further includes additional purification steps to obtain a final virus preparation containing infectious virus particles while controlling impurities and aggregates. Size-exclusion techniques are referenced as optional, including sucrose density gradient centrifugation and size exclusion modalities, and the overall concept is to enrich and purify infectious virus particles while addressing impurities generated from cell substrate and limiting residual host cell DNA, residual host cell protein, and virus particle aggregates.
Claims Coverage
The independent claims are directed to protamine-based separation and purification of infectious virus particles from preparations containing non-infectious virus particles and impurities, limited to specified virus families and expressly excluding Zika virus particles. Two inventive features are identified.
Protamine precipitation separation excluding Zika
Precipitating non-infectious virus particles with protamine to separate infectious virus particles from non-infectious virus particles, for virus particles selected from Paramyxoviridae, Orthomyxoviridae, Flaviviridae, Filoviridae, Arenaviridae, Rhabdoviridae, Togaviridae and Coronaviridae, wherein the virus particles are not Zika virus particles.
Protamine-contact purification with defined infectious enrichment range excluding Zika
Providing a crude harvest comprising infectious virus particles, non-infectious virus particles, and impurities generated from growing the virus particles on a cell substrate; contacting the crude harvest with an agent comprising protamine to obtain a virus preparation comprising infectious virus particles, wherein the enrichment of infectious virus particles relative to total virus particles in the crude harvest is at least 50% to 95%, for virus particles selected from Paramyxoviridae, Orthomyxoviridae, Flaviviridae, Filoviridae, Arenaviridae, Rhabdoviridae, Togaviridae and Coronaviridae, wherein the virus particles are not Zika virus particles.
Across the independent claims, the core claimed inventive concept is protamine-based precipitation/contact to enrich infectious virus particles from preparations that contain both infectious and non-infectious virus particles and impurities, limited to specified virus families and expressly excluding Zika virus particles.
Stated Advantages
Enriches infectious virus particles relative to total virus particles in the crude harvest in the range of at least 50% to 95%.
Separates infectious virus particles from non-infectious virus particles by precipitating non-infectious virus particles with protamine.
Facilitates separation of infectious virus particles from host cell proteins and/or low molecular weight materials.
Optionally enables further purification using size-exclusion techniques while controlling residual host cell DNA, residual host cell protein, and virus particle aggregates.
Documented Applications
No documented applications found
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