The 20th annual Symposium will be held on Thursday, April 23, 2009.

Chromatographic Purification of Recombinant Adeno-Associated Viral Vectors Using Convective Interaction Media
Norris 100, 3:15 PM

Man Kai (Alyssa)  Wong,   '09 17
   Majors: Chemistry, Music
   Hometown: Hong Kong, China

Sponsor(s): Vanessa McCaffrey
Support: Cystic Fibrosis Foundation. This research was conducted at University of Pennsylvania School of Medicine under the guidance of Dr. Martin Lock

Abstract: 
Adeno-associated virus (AAV) is one of the most promising viral vectors for human gene therapy. The classic method of AAV purification by density gradient centrifugation is only effective for a small scale. A scalable production strategy of rAAV is often demanded for the use of clinical trials with larger animals. The objective of the study is to generate a scalable and robust rAAV purification system using ion exchange chromatography. For this study AAV serotype 7 and 6.2 were chosen. For optimal resolution and elution of AAV, a wide range of salt concentrations and buffer pH was screened. A set of optimal conditions was designed based on the results of initial screening that allows maximum yield of the purified vector with minimal loss of viral protein in flow through. The resolution of the two AAV serotypes was studied using a convective interaction media column (CIM-QA), which has large surface area for a flow independent resolution and binding capacity. The result demonstrated that a considerable amount of the AAV7 viral protein could be recaptured from the column in a single step, making AAV7 a favorable candidate for monolith chromatography. AAV6.2 showed slight resolution, and with a significant loss in the flow through. This chromatography method may offer advantages in ease of purification, final vector purity, and process scalability. Large-scale production of AAV is possible with careful selection of column media and elution conditions. The optimal pH and salt concentration for purification of different AAV serotypes may vary, and may affect the overall yield of the vector.

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