Patent ID: 8574583

Claim:
In a method of screening an AAV viral library comprising hybrid full-length capsid genes prepared by: (i) isolating AAV capsid nucleotide sequences from two or more AAV serotypes; (ii) digesting the AAV capsid nucleotide sequences into fragments; (iii) reassembling the fragments into a full-length gene using primer-less PCR to form PCR products; (iv) cloning the re-assembled PCR products into plasmids to generate a library of recombinant AAV plasmids, an improvement for increasing infectivity of an AAV virion and enhancing resistance to neutralization by anti-AAV antibodies, said improvement comprising: (a) transfecting mammalian cells with the library of recombinant AAV plasmids from step (iv); (b) incubating the transfected cells in the presence of anti-AAV antibodies until the cells lyse to produce an amplified and packaged AAV viral library; and (c) passaging the AAV viral library in mammalian cells in the presence of anti-AAV antibodies and selecting an AAV virion that survives said passaging, wherein the selected AAV virion has increased infectivity and enhanced resistance to neutralization by anti-AAV antibodies, relative to wildtype AAV-2.