利用肝素化的纖維素基質吸附第二型重組腺病毒並進行局部基因傳導

Application of Heparinized Cellulose Affinity Membranes in Recombinant Adeno-Associated Virus Serotype 2 Binding and Delivery

王志豪(Zhi-Hao Wang)

指導教授 ： 湯學成

國立清華大學/工學院/化學工程學系/碩士(2008年)

https://doi.org/10.6843/NTHU.2008.00192

摘要

The microporous affinity membrane based on cellulose matrices offers minimal mass-transfer effects in membrane chromatography with low nonspecific adsorption. In this
research, we tested a novel application of the microporous, heparinized cellulose matrices (H-CM) for their affinity toward recombinant adeno-associated virus serotype 2 (rAAV2,
which uses heparan sulfate proteoglycans as the primary cellular receptor) to develop a controlled, substrate-mediated viral vector delivery. We conjugated rAAV2 to an
epoxy-crosslinked heparin cellulose membrane, which led to vector transduction upon cellular adhesion. When adhered, the human HT-1080 fibroblasts exhibited proliferation
kinetics similar to those on the standard polystyrene tissue-culture surface. Using green fluorescent protein and β-galactosidase as reporters, we showed that the heparin-bound rAAV2 particles remained active and that the rAAV2-heparin binding was reversible and capable of mediating transgene delivery in cell culture. In addition, we applied the affinity membrane to adsorb unpurified rAAV2 from the crude lysate of packaging cells via the ligand-receptor binding, avoiding the use of conventional ultracentrifugation or chromatography in preparation of infectious rAAV2 for transduction. Our work explores a
new application of affinity cellulose matrices in substrate-mediated viral vector delivery, which can be a useful tool in developing protocols for localized gene transfer.

關鍵字

第二重組腺病毒 ； 親和性層析法 ； 纖維素 ； 基因傳導 ； 肝素

並列摘要

無資料

並列關鍵字

AAV2 ； affinity membrane chromatography ； cellulose ； gene transfer ； heparin

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