Recombinant adeno-associated virus particle purification comprising an affinity purification step

A method for obtaining purified recombinant Adeno-Associated Virus (rAAV) particles, comprising the steps of:

• a) treating a starting material previously obtained from cells producing rAAV particles with a detergent, said starting material comprising a cell lysate and/or a culture supernatant;
• b) performing a depth filtration of said a starting material using a depth filter membrane comprising borosilicate glass microfibers and mixed esters of cellulose, whereby a rAAV-containing clarified composition is provided;
• c) submitting the rAAV-containing clarified composition to an immunoaffinity purification step and eluting the rAAV particles at acidic pH, whereby a first rAAV-enriched composition is provided;
• d) neutralizing the acidic pH of said first rAAV-enriched composition of step c) to neutral or basic pH;
• e) submitting the first rAAV-enriched composition of step d) at least once to:
• e1) a step of anion-exchange chromatography on a chromatographic support, wherein elution is performed by using a salt gradient and wherein the rAAV-containing fraction is collected, whereby a second rAAV-enriched composition is provided;
• or
• e2) a step of density gradient centrifugation, wherein the rAAV-containing fraction is collected, whereby a second rAAV-enriched composition is provided;
• f) submitting the second rAAV-enriched composition to a step of tangential flow filtration, whereby purified recombinant Adeno-Associated Virus (rAAV) particles are provided,
• wherein the method does not comprise an apatite chromatography step or a cation-exchange chromatography step, and
• wherein the rAAV particles belong to an AAV serotype selected from the group consisting of AAV4, AAVrh10, AAV2, AAV8, and AAV9.