Purification of Functional Gene Transfer Agents Using Two-Step Preparative Monolithic Chromatography

Background: Gene transfer agents (GTAs) are phage-like particles that transfer cellular genomic DNA between cells. A hurdle faced in studying GTA function and interactions with cells is the difficulty in obtaining pure and functional GTAs from cultures. Materials and Methods: We used a novel two-step method for purification of GTAs from R. capsulatus by monolithic chromatography. Results: Our efficient and simple process had advantages compared to previous approaches. The purified GTAs retained gene transfer activity and the packaged DNA could be used for further studies. Conclusions: This method is applicable to GTAs produced by other species and small phages, and could be useful for therapeutic applications.