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| 題 名 | Preparation of Glutathione-S-Transferase Piezoelectric Immunosensors with Hydrogel-Coating=水膠塗覆製備Glutathione-S-Transferase壓電免疫感測器之研究 |
|---|---|
| 作 者 | 陳秀美; 林裕勝; | 書刊名 | Journal of the Chinese Institute of Chemical Engineers |
| 卷 期 | 30:5 1999.09[民88.09] |
| 頁 次 | 頁413-420 |
| 分類號 | 460.02 |
| 關鍵詞 | 水膠塗覆; 壓電免疫感測器; 日本血吸蟲; Hydrogel-coating; Glutathione-S-Transferase; Immunoassay; Piezoelectric; Protein A; Schistosomiasis; SjGST; |
| 語 文 | 英文(English) |
| 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與共抗體間之作用符合Langmuir吸附動力模式,而所測得 之K□值為8x10□M□。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酵素及共突變種。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 L-lysine modified piezoelectric crystal surface, a reusable Schistosoma japonicum glutathione-Stransferase (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 value of 8X 10 □M□ 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 ani:ibody 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 irnmunoassay 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. |
本系統中英文摘要資訊取自各篇刊載內容。