A scalable manufacturing platform for viral vector purification and viral vectors so purified for use in gene therapy

A scalable manufacturing platform for viral vector purification and viral vectors so purified for use in gene therapy

  • CN 102,947,453 A
  • Filed: 01/25/2011
  • Published: 02/27/2013
  • Est. Priority Date: 01/28/2010
  • Status: Active Application
First Claim
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1. one kind is used for comprising the genetically modified real AAV carrier granule that coding is treated albumen or its fragment from the AAV prepared product purifying that comprises AAV carrier granule, hollow capsid and host cell impurity, thereby the method for the AAV product that does not substantially contain the AAV hollow capsid is provided, and described method comprises:

  • A) results comprise the cell of the AAV that recombinates;

    B) via the concentrated described cell of tangential flow filtration;

    C) pass through the described cell of microjet cracking to form lysate;

    D) filter, thereby make step c) the lysate clarification;

    E) by ion-exchange chromatography method purifying AAV particle and optional by tangential flow filtration evaporating column elutriant;

    F) mix described elutriant and cesium chloride and carry out described mixture centrifugal;

    G) at step f) collect afterwards virion and carry out identical buffer-exchanged by tangential flow filtration;

    H) prepare the AAV particle of purifying so that the AAV granular preparation to be provided with tensio-active agent;

    I) filter described preparation to remove any residual impurity, the amount of the described real AAV carrier granule that exists in the wherein said AAV product is at least 95%.

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