429. Alpha 1 Antitrypsin Gene Delivery for Preventing Type 1 Diabetes in NOD Mice

Abstract

We have created a chemical strategy for the preparation of stable conjugates of functional AAV (adeno-associated viral vectors) with solid microbeads (nanocarriers). The conjugation of AAV to the microbeads used the streptavidin-biotin system, which provides a very strong and stable association between the viral particle and the microbead. Optimal biotinylation conditions were initially determined, under which biotin groups can be attached covalently to the outer surfaces of AAV particles without disturbing their intrinsic ability to bind and infect cells. The biotinylated AAV particles can be conjugated efficiently to the surfaces of streptavidin-coated polystyrene microbeads. The ability of these AAV-microbead conjugates to infect cells was investigated by using two strategies. One strategy involved the co-attachment of lectins to the microbead surface. This should provide the conjugates with the ability to bind to carbohydrate moieties on the cell surface. The other strategy used streptavidin-coated polystyrene microbeads containing a superparamagnetic core as virus carriers. This should allow the conjugates to associate tightly with target cells by using magnetic force. The use of these two strategies allowed efficient infection of target cells by conjugated AAV particles, indicating that conjugated AAV particles are capable of infecting target cells efficiently upon stable association of AAV-microbead conjugates with the cell surface. Thus, AAV-microbead conjugates offer great potential to be used for the development of methods for enhanced delivery of AAV to specific sites or tissues in a controlled, versatile manner.