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|標題:||The use of matrix in dye-immunochromatographic application
|作者:||賴信方||關鍵字:||immunochromatographic test;免疫層析檢測;dye immunochromatographic test;dextran;BSA;polylysine;染料免疫層析檢測;葡萄聚糖;牛血清白蛋白;聚離胺酸||出版社:||化學工程學系||摘要:||
Dye immunochromatographic test（DICT）which can be used in multi-colored test lines on test strips offers the advantage of easier distinction in diseases triage and wider analyte concentration ranges in quantitative as compared to the conventional one-colored immunochromatographic test（ICT）. In this study, we attempt to dye the matrix such as dextran, BSA, polylysine, as the marker to label antibody for appling in DICT.
In this reserch, we divided the results into three parts：
（1）Dextran：The optimal dying the dextran conditions were found by the RSM（response surface methodology）as follows; 40.23 ℃、pH 11.09、16.84 mg dye/0.5mg dextran、3.5hr. Then we used the dye-dextran to label antibody. No colore was detected in the test line. It is likely that the dying rate is too low or the antibody is denatured during the conjugated reaction.
（2）BSA：BSA was dyed to the saturated extent. The conditions for employing the dye-BSA to label antibody to obtain the deepest color in the test strips were : pH 6.5、molar ratio (mole dye/mole antibody) : 10. Taking the HSA competition assay as examples, HSA could be measured within the linear ranges from 0 to 18.92 μg/ml.
（3）Polylysine：As compared the results of different conditions on dying polylysine, the best conditions for preparing dye-polylysine were : molecular weight 189.4k Da、pH 11、40℃、molar ratio (mole dye/mole amino group) : 1.5. Anti-HSA labeled with dye-polylysine was used to get the deepest color in the test strips. Taking the HSA competition assay as examples, HSA could be dedected within the linear ranges from 0 to 10.44 μg/ml.
在本論文中，將反應性染料（PROCION BLUE MX-7RX）與不同介質進行反應，再將此染色介質用以標誌抗體，並應用於免疫層析檢測，此方式稱為染料免疫層析檢測（Dye immunochromatographic test, DICT）。以多株抗體（polyclonal anti-HSA）及抗原（HSA）為檢測模式蛋白，探討三種染色介質（即染色dextran、染色BSA、染色polylysine）與抗體鍵結之顯色結果，並進一步探討染料免疫層析檢測在定量免疫分析上之應用。
（1）Dextran：利用回應曲面法來求染色dextran之最適條件：40.23℃、pH 11.09、16.84 mg染料/0.5mg dextran、3.5hr，並以此標誌抗體，但因染色率太低及鍵結反應的過程中造成抗體失活，故檢測試紙並無顯色。
（2）BSA：將BSA染色至飽和程度，利用1-Ethyl-3-(3-dimethyl aminopropyl)carbodimide（EDC）將染色BSA標誌抗體，於檢測試紙顯色有較佳條件為：染色BSA對抗體莫耳比10、pH 6.5，再用此已標誌抗體進行競爭型檢測，檢測線性範圍為0~18.92μg/ml。
（3）Polylysine：當染色polylysine製備條件為：分子量189.4k Da、pH 11、40℃、染料對胺基莫耳比1.5，此染色polylysine標誌抗體後，於檢測試紙上有最佳顯色結果，再用此已標誌抗體進行競爭型檢測，檢測線性範圍為0~10.44μg/ml。
|Appears in Collections:||化學工程學系所|
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