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RECOMBINANT ADENO-ASSOCIATED VIRUS PARTICLE PURIFICATION COMPRISING AN AFFINITY PURIFICATION STEP
专利权人:
CHU NANTES;UNIVERSITE DE NANTES;ASSOCIATION FRANCAISE CONTRE LES MYOPATHIES;INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICAL (INSERM)
发明人:
Nicole BRUMENT
申请号:
US15549186
公开号:
US20190382733A1
申请日:
2016.02.09
申请国别(地区):
US
年份:
2019
代理人:
摘要:
The invention describes a method for obtaining purified recombinant Adeno-Associated Virus particles (rAAV), comprising the steps of: a) performing a depth filtration of a starting material previously obtained from cells producing rAAV particles, the said starting material being selected in a group comprising a cell lysate and a culture supernatant, whereby a rAAV-containing clarified composition is provided; b) submitting the rAAV-containing clarified composition to an affinity purification step, whereby a first rAAV enriched composition is provided; c) submitting the first rAAV enriched composition at least once to: c1) a step of anion-exchange chromatography on a chromatographic support wherein elution is performed by using a salt gradient, preferably a linear salt gradient, and wherein the rAAV-containing fraction is collected, whereby a second rAAV enriched composition is provided; or c2) a step of density gradient centrifugation, wherein the rAAV-containing fraction is collected, whereby a second rAAV enriched composition is provided; d) submitting the second rAAV enriched composition to a step of tangential flow filtration, whereby purified recombinant Adeno-Associated Virus particles (rAAV) are provided.
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