發展快速單一步驟免疫層析法檢測血液中低密度脂蛋白濃度之研究

Abstract

動脈硬化塊的組成，主要是以血脂肪（Lipid core）為中心，其成分主要是膽固醇，包在周圍的是平滑肌細胞和各種結締組織，一般檢驗是否為心臟病的高危險群是以血液中膽固醇的濃度作為判斷的根據，而又以低密度脂蛋白中的膽固醇濃度最具指標性。 本實驗最主要的目的是結合金膠及低密度脂蛋白單株抗體，利用免疫層析法，發展出一套快速檢驗血液中低密度脂蛋白濃度的檢測試劑，因為一般檢測血液中低密度脂蛋白的方法主要為ELISA，但ELISA耗費的時間很長且步驟複雜不適合居家檢測，所以為了達到快速檢測且適合居家檢測的目的，本實驗室先製造出四株LDL單株抗體，再利用實驗合成之奈米金顆粒 (gold nanoparticles) 相結合形成免疫金膠(immunogold)，應用於快速單一步驟之免疫層析法(immunochromatographic assay) ，研發出只需1 μL的檢體且2分鐘即可完成檢測的試劑，並期望此檢測試劑未來能進一步發展成居家即可檢測血液中低密度脂蛋白濃度的試劑，達到預防動脈硬化之目標。

Atherosclerosis involves deposits of fatty substances, cholesterol, cellular metabolites, calcium and other extracellular matrix in the lining of an artery. High cholesterol is a leading risk factor in causing heart disease. Excess cholesterol in plasma low-density lipoprotein (LDL) produces plaque (a thick, hard deposit) in arterial wall. The purpose of this study was to develop a rapid immunochromatographic assay using colloidal-gold conjugated monoclonal antibodies against LDL. Among which ELISA is the most popular technique for the detection of LDL concentration and is highly sensitive. Due to the time-consuming of ELISA, development of a fast, accurate and easy-to-use home test is necessary to monitor the LDL. For this reason, 4 lines of LDL monoclonal antibodies were produced and prepared. The method was able to determine the LDL moiety of human plasma within minutes without tedious ELISA procedures. In general, 1 ul of plasma was enough to complete the assay. We anticipate to produce a home-test kit to determine LDL concentration in the future.