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題 名 | 應用馬來醯亞胺法製備酶聯結抗體以偵測竹嵌紋病毒=Preparation of Rabbit IgG-Alkaline Phosphatase Conjugate by Maleimide Method and Its Application for Detecting Bamboo Mosaic Potexvirus |
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作 者 | 陳滄海; 盧耀村; | 書刊名 | 國立屏東科技大學學報 |
卷 期 | 9:2 2000.06[民89.06] |
頁 次 | 頁125-132 |
分類號 | 433.4 |
關鍵詞 | 抗體酵素標識法; 馬來醯亞胺法; 竹嵌紋病毒; Enzyme-labeling of antibody; Maleimide method; Bamboo mosaic potexvirus; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究以馬來醯亞胺法(maleimide method)作竹嵌紋病毒(bamboo mosaic potexvirus, BaMV)抗體之鹼性磷酸?標識(labelling),並以DAS-ELISA評估其與傳統一段式戊二醛法在抗體酵素標識效益之差異。純化之免疫球蛋白經S- acetylmercaptosuccinic anhydride處理及Sephadex G-25層析而硫酐化(mercaptosuccinylation)後,與馬來醯亞胺化之鹼性磷酸?作用再經管柱層析即可得鹼性磷酸?標識BaMV抗體,層析過程中,比較Sepharose CL-6B,Sephadex G-100,及Ultragel AcA34三種不同凝膠,結果以Sephadex G-100層析分離效果最佳且所得?聯結抗體在進行DAS-ELISA時貸反應靈敏度高出目前常用之一段式戊二醛法所製備之聯結抗體5.5倍,其有效偵測BaMV下限濃度可達10ng/ml。 |
英文摘要 | Antibody against bamboo mosaic potexvirus (BaMV) was conjugated with alkaline phosphatase by maleimide method and the conjugate obtained was compared with that of the conjugate prepared by one step glutaraldehyde method in sensitivity evaluated with DAS-ELISA. Purified rabbit anti-BaMV IgG was first mercaptosuccinylated in the presence of S-acetylmercaptosuccinic anhydride, then was coupled to alkaline phosphatase using N-succimidy 1-4-(N-maleimidomethyl) cyclohexane-1-carboxylate, finally was successively separated with three chromatography gels, i.e. Sepharose CL-6B, Sephadex G-100 and Ultragel AcA34. The Sephadex G-100 showed the better separation effect than the other two gels. The enzyme-conjugated immunoglobulin prepared by maleimide method was 5.5 fold more sensitive than that of the conjugated immunoglobulin prepared by one step glutaraldehyde method. It could detect BaMV at the concentration of 10ng/ml. |
本系統中英文摘要資訊取自各篇刊載內容。