免疫層析試紙測試法組裝最適化及對streptavidin蛋白質之快速偵測

Optimization of An Immunochromatographic Assembly and Rapid Detection to Streptavidin Protein

吳凱智(Kai-Chih Wu)

指導教授 ： 吳瑞璋

中原大學/工學院/化學工程學系/碩士(2007年)

https://doi.org/10.6840/cycu200700516

摘要

摘要:
免疫層析試紙分析法(Immunochromatography)為現代快速生物感測重要方法之一，其原理是藉由抗體抗原間的專一性與免疫親和力做檢測，被普遍應用在臨床檢測、海關、法醫、警局臨檢驗毒等方面。
在目前的研究中多為定性或半定量檢測；另外免疫層析試紙分析法在顯色標定物上也有許多不同，大多使用乳膠（latex）、膠體金（colloid gold）與酵素（enzyme），而在文獻中也有使用染料（dye）、及碳黑（carbon black）。
本實驗依免疫層析分析法之原理，藉由毛細引力（capillary attraction）使待測物(蛋白質streptavidin)在nitrocellulose薄膜中移動，藉待測抗原-抗體mouse anti-streptavidin與報告抗體biotinylated mouse anti-human IL6間的免疫反應(immunoreaction)，經由膠體金顯色，檢測出抗原之存在。本研究除完成組裝與試劑參數最適化之外，也測試出抗原含量之偵測極限(detection limit)在1pg/ml左右，此結果與所見文獻中偵測極限約10-12g/ml相同[1]。

關鍵字

側向流檢測試紙 ； 免疫層析檢測 ； 薄膜快速偵測

並列摘要

Abstract:
The immunochromatographic test （ICT）is one of the important biosensing techniques. The principle of ICT bases on the immunoaffinity partition of the analyte among antibodies and antigen. ICT has been widely used at clinical, custom house, and the public security bureau.
In present research, the multi-analyte assay is qualitative or half- quantitative. In addition, it has many different types of chromophors as ICT visualization markers. In common usage, they are latex, colloid gold, enzyme, dye, and carbon black as mentioned in the literature.
In this study, immunochromatographic test is adopted to detect the analyte (streptavidin), which migrates laterally through the porous NC membrane by the capillary force. The immunoreaction between mouse anti-streptavidin and biotinylated mouse anti-human IL6 is observed via the colloidal gold labeling.
This research successfully assembed the test device and performed the detection of streptavidin. The detection limit was about 1pg/ml, same as the report in the literature of 10-12g/ml. [1]

並列關鍵字

rapid test ； immunochromatographic assay ； lateral flow

參考文獻

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