EDC/NHS-Mediated Heparinization of Small Intestinal Submucosa for Recombinant Adeno-Associated Virus Serotype 2 Binding and Transduction

• Main Authors: Tse-Wei Yue, 余則威
• Other Authors: Shiue-Cheng Tang
• Format: Others
• Language: en_US
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/72232177107904400270

碩士 === 國立清華大學 === 化學工程學系 === 95 === A major challenge in the use of gene transfer vectors as therapeutic tools is controlling vector administration at a desired tissue site. One potential solution is implanting tissue-engineering constructs loaded with gene transfer vectors such as viruses for localized transgene delivery. In this work, we conjugated recombinant adeno-associated virus serotype 2 (rAAV2) to a heparinized small intestinal submucosa (H-SIS) matrix, which resulted in vector transduction upon cellular adhesion. H-SIS was prepared by incorporating heparin, the rAAV2 receptor, into SIS through N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) mediated crosslinking. Incorporated heparin adsorbed rAAV2 onto the H-SIS matrix for conjugation. Using green fluorescent protein and ��-galactosidase as reporters, we showed that conjugated rAAV2 was active and capable of mediating transgene delivery in cell culture. Our work provides a unique, modified tissue substrate H-SIS for rAAV2 binding and transduction, which can be a useful tool in developing localized gene transfer.