Development of New Assays for Lipoprotein Testing

脂蛋白分数分析是评估冠状动脉疾病风险的一种有价值的工具。然而，当前的程序需要多次测试，并且有几个手动步骤。为了降低脂蛋白分析的复杂性和成本，我们开发了一种单管均质分析方法，用于测量血清高密度脂蛋白（HDL）胆固醇、总胆固醇和甘油三酯。然后可以使用弗里德瓦尔德方程从这些参数计算低密度脂蛋白（LDL）胆固醇。该试验使用抗载脂蛋白b抗体来阻断报告酶对低密度脂蛋白胆固醇的反应性。测定以顺序的方式进行，以便在确定高密度脂蛋白胆固醇后，添加洗涤剂以破坏抗体复合物，从而允许随后的总胆固醇测量。接下来，添加用于测量总甘油三酯的报告酶。在接下来的一年里，我们计划测试该方法与其他阻断低密度脂蛋白胆固醇反应性的方法的兼容性。我们还计划开发低密度脂蛋白胆固醇的同质分析，随后是总胆固醇和甘油三酯的顺序分析。

Lipoprotein fraction analysis is a valuable tool in estimating the risk for coronary artery disease. The current procedure, however, requires multiple tests and has several manual steps. In order to reduce the complexity and cost of lipoprotein fraction analysis, we have developed a single-tube homogenous assay for measuring serum high-density lipoprotein (HDL) cholesterol, total cholesterol, and triglyceride. Low-density lipoprotein (LDL) cholesterol can then be calculated from these parameters using the Friedewald equation. The assay uses an anti-apoB antibody to block the reactivity of the reporter enzymes to LDL-cholesterol. The assay is performed in a sequential manner so that after the HDL-cholesterol is determined, a detergent is added to disrupt the antibody complex, which allows the subsequent measurement of total cholesterol. Next, the reporter enzymes for measuring total triglyceride are added. In the upcoming year, we plan to test the compatibility of the method with alternative approaches for blocking the reactivity of LDL-cholesterol. We also plan to develop a homogenous assay for LDL-cholesterol, followed by sequential assays for total cholesterol and triglyceride.