Multimodal metal affinity processing AAV capsids

Abstract

A method for separating full Adeno-associated virus (AAV) capsids from empty AAV capsids in a buffered mixture comprising full AAV capsids, empty AAV capsids, comprising the steps of contacting the buffered mixture with a first substrate bearing a metal affinity ligand attached to the first substrate, said metal affinity ligand having the ability to complex metal ions via three or more nitrogen atoms, separating empty AAV capsids from full AAV capsids by eluting with a pH gradient, a salt gradient, a metal ion gradient or a combination thereof in the presence of multivalent cations bound to the metal affinity ligand to obtain a purified full AAV capsid fraction. For removing contaminating DNA in the mixture or purified AAV capsid fraction, the method of the invention can be combined with contacting of the buffered mixture or the purified full AAV capsid fraction with a second substrate bearing a metal affinity ligand attached to the second substrate in the presence of multivalent cations bound to the metal affinity ligand, said metal affinity ligand comprises two or more negatively charged carboxylic acid residues.

Dependencies are inferred from the claim text.