AAV capsid library and AAV capsid proteins

1. 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.

2. The improvement of claim 1 , wherein isolating includes isolating AAV capsid nucleotide sequences from human AAV serotypes and non-human AAV serotypes.

3. The improvement of claim 2 , wherein isolating includes isolating AAV capsid nucleotide sequences selected from the group consisting of AAV-2, AAV-8, and AAV-9.

4. The improvement of claim 1 , wherein said transfecting comprises transfecting into 293 kidney cells with a helper virus.

5. The improvement of claim 1 , wherein said anti-AAV antibodies comprise human anti-AAV antibodies.

6. A recombinant AAV (rAAV) library prepared according to the improvement of claim 1 .