AAV capsid library and AAV capsid proteins

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.