Sialyltransferase Inhibitor-Nanoparticle Conjugates for Diagnosis, Separation, and Enrichment of Functional Proteins

• Main Authors: Wei-Jen Chiang, 江維恁
• Other Authors: Wen-Shan Li
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/62101878992995913512

碩士 === 國立中央大學 === 化學研究所 === 98 === Nanoparticles bearing surface-conjugated specific inhibitors are increasingly being utilized for a number of bio-applications including identification, separation and enhancement of desired proteins. These attractive bio-applications involve specific adsorption and recognition; however, these are the major hindrances for nanobiotechnology. To solve this question, we have investigated the effect of nanoparticle surface displaying high affinity molecules, such as sialyltransferase inhibitors. Maintaining a reasonable affinity toward desired protein is generally a prerequisite for proper design of nanoparticle-conjugated specific inhibitors. We prepared sialyltransferase inhibitors, lithocholic acid derivatives with L-Asp or NBD-L-Asp moiety, which have IC50 values at micromolar ranges. Next, magnetic nanoparticle (iron oxide)-conjugated lithocholic acid derivatives, compounds 15~16, were synthesized via click chemistry. Herein, we demonstrated that compounds 15~16 have the ability to identify, separate and enhance binding to the target alpha-2,3(N)-sialyltransferase, a glycol- and membrane-bound protein. The protein band (38 kDa) on SDS-PAGE derived from rat was confirmed by LC MS-MS analysis and proteomics searching. Extending these studies to crude cell lysate in vitro experiment, we found several interesting proteins including talin. Further investigation of talin toward metastasis in cancer is in progress.