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REINIGUNG VON REKOMBINANTEN, ADENO-ASSOZIIERTEN VIREN MIT EINEM IMMUNAFFINEN REINIGUNGSSCHRITT
专利权人:
INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE (INSERM);UNIVERSITE DE NANTES;CHU Nantes;Association Francaise Contre Les Myopathies (AFM)
发明人:
申请号:
EP15305188.3
公开号:
EP3054007A1
申请日:
2015.02.09
申请国别(地区):
EP
年份:
2016
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 immunoaffinity 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; orc2) 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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