RECOMBINANT ADENO-ASSOCIATED VIRUS PARTICLE PURIFICATION COMPRISING AN AFFINITY PURIFICATION STEP

1 - 16 . (canceled)

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

i. performing a depth filtration of a starting material previously obtained from host cells producing rAAV particles, said starting material comprising a cell lysate and/or a cell culture supernatant, whereby a rAAV-containing clarified composition is provided;

ii. submitting the rAAV-containing clarified composition to an immunoaffinity purification step, whereby a first rAAV enriched composition is provided;

iii. 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, 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;

iv. submitting the second rAAV enriched composition to a step of tangential flow filtration, whereby purified rAAV particles are provided.

18 . The method according to claim 17 , wherein step b) is performed by using a chromatography support onto which antibodies or fragments thereof directed to the said rAAV particles, are immobilized.

19 . The method according to claim 18 , wherein the antibodies are monoclonal.

20 . The method according to claim 18 , wherein the antibodies are camelid antibodies.

21 . The method according to claim 17 , wherein step b) is performed using an AVB Sepharose™ chromatography (GE Healthcare) support.

22 . The method according to claim 17 , wherein the at least one chromatographic support at step c1) is a monolithic chromatographic support.

23 . The method according to claim 17 , wherein elution is performed by using a linear salt gradient.

24 . The method according to claim 17 , wherein step d) is performed by using a filter membrane having a molecular weight cut-off value ranging from 20 kDa to 150 kDa.

25 . The method according to claim 17 , wherein the rAAV particles have an AAV capsid belonging to a serotype selected in a group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV8, AAV9, AAV10, and rhesus macaque-derived serotypes, and mixtures thereof.

26 . The method according to claim 25 , wherein the rhesus macaque-derived serotype is AAVrh10.

27 . The method according to claim 17 , wherein the rAAV particles have an AAV capsid belonging to the AAV5 serotype, and wherein the rAAV particle contains DNA comprising an expression cassette encoding human PDE6-beta.

28 . The method according to claim 17 , wherein the rAAV particles have an AAV capsid belonging to the AAV4 serotype, and wherein the rAAV particle contains DNA comprising an expression cassette encoding human RPE65.

29 . The method according to claim 17 , wherein the purified rAAV particles are suitable for use for gene therapy.

30 . The method according to claim 17 , wherein the purified rAAV particles have a purity of at least 90%.

31 . The method according to claim 17 , wherein the purified rAAV particles have an amount of residual host cell DNA equal or inferior to 50 ng per dose of 10 12 vector genomes (vg).

32 . A rAAV particle obtained by the method according to claim 17 .

33 . A composition comprising the rAAV particle according to claim 32 .

34 . The composition according to claim 33 , comprising rAAV particles with a purity of at least 90% and/or an amount of residual host cell DNA equal or inferior to 50 ng per dose of 10 12 vector genomes (vg).

35 . A host cell transfected with an rAAV particle according to claim 32 .

36 . A method of treating a disease in a subject in need thereof by gene therapy, comprising administering to said subject the rAAV particle according to claim 32 comprising a heterologous nucleic acid encoding a gene product, or a composition comprising the rAAV particle.