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。在过去的一年里，我们已经在标准的临床化学分析仪上实现了完全自动化的分析，美国国立卫生研究院已经将这项技术授权给了一家外部公司，以进行潜在的商业化。在即将到来的一年里，我们计划进一步改进该分析方法，并计划评估该分析方法被许可方开发的任何商业测试。我们还计划研究一种新的比色脂肪酶底物用于常规诊断脂肪酶检测。

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 past year, we have fully automated the assay on a standard clinical chemistry analyzer, and the NIH has licensed the technology to an outside company for potential commericialization. In the upcoming year, we plan to make further refinements of the assay and plan to evaluate any commercial tests developed by the licensee of the assay. We also plan to investigate the use of a new colorimetric lipase substrate for routine diagnostic lipase testing.