• 作者： Chen, Hsiu-Mei; Lin, Yu-Sheng
• 中文摘要： 利用一簡易的水膠塗覆技術, 以氧化葡聚醣為架橋將蛋白質固定於Poly-L-lysine修飾過之電極表面, 可穩定地製備固定有日本血吸蟲Glutathione-S-transferase (SjGST)之壓電免疫感測晶片。研究結果顯示, 所製備之感測晶片, 其特異性極佳, 而且可穩定地對其抗體進行連續重覆檢測。固定化SjGST與其抗體間之作用符合Langmuir吸附動力模式, 而所測得之K/sub a/值為8*10/sup 7/M/sup -1/。然而, 由於SjGST快速失活之故, 所製備晶片在一天後即活性下降。此外, 以相同方法製備固定有Protein A之感測晶片, 可親和固定化 SjGST之抗體, 並進而間接免疫檢測SjGST酵素及其突變種。利用此方法可正確地固定抗SjGST抗體之方向, 使得平均每個抗體可和1.9個 SjGST單體分子結合。此研究結果證明以壓電晶片免疫檢測SjGST及其抗體之可行性。此技術不僅可應用於血吸蟲病之檢測, 還可用於其它以GST為生物指標之疾病檢測(如細胞壞死及癌症)。
• 英文摘要： By using a simple hydrogel-coating technique, where a periodate-oxidized dextran matrix was used as a double- sided linker to covalently immobilize protein molecules onto a poly-L-lysine modified piezoelectric crystal surface, a reusable Schistosoma japonicum glutathione-S- transferase (SjGST)-coated sensing crystal was prepared. The sensing crystal was highly specific and stable in continuos repetitive immunoassay of anti-SjGST antibody. The interaction of anti-SjGST antibody with immobilized SjGST followed the Langmuir single-site adsorption and a k/sub a/ value of 8*10/sup 7/M/sup -1/ was obtained. However, the prepared crystal rapidly lost its activity in one day because of the fast denaturation of SjGST. In addition, a protein A-coated crystal was also prepared by using the same hydrogel-coating technique to affinity- capture anti-SjGST antibody for the subsequent immunoassay of SjGST and its genetically engineered mutant. The antibodies affinity-captured by protein A were properly oriented such that on average each antibody bound about 1.9 molecules of the monomer of SjGST under the experimental conditions. This study demonstrated the feasibility of the immunoassay of SjGST or anti-SjGST antibody for Schistosomiasis by using piezoelectric crystals. The similar technique can be applied to the immunoassay of other GSTs that are biomarkers of organ damage or cancer risk.
• 中文關鍵字： 水膠塗覆; 免疫分析; 壓電; 蛋白質
• 英文關鍵字： Hydrogel Coating; Immunoassay; Piezoelectric; Protein