METHOD FOR ISOLATION AND MICROASSAY OF LIPOPROTEIN A

脂蛋白A的分离和微量测定方法

• 批准号: 2702299
• 负责人: Mary Gaunt-Kloepfer
• 金额: $ 28.37万
• 依托单位: MICRONIX, INC.
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2702299
• 关键词: affinity chromatography; blood lipoprotein; blood tests; cholesterol; diagnosis design /evaluation; lectin; method development; physical separation; rapid diagnosis; serology /serodiagnosis

It has been postulated that lipoprotein (a) [Lp(a)] may be the most atherogenic lipoprotein in man. Current laboratory methods for Lp(a) employ immunoquantitation [apo(a)]. Because these methods are hampered by cross-reactivity to plasminogen, apo(a) isoform dependence, antibody heterogeneity, and lack of standardization, none of them is FDA-approved. One third of Lp(a)'s mass is cholesterol, and only one tenth is apo (a). We propose develop a quantitive affinity-chromatographic, and a semiquantitative teststrip methods for Lp(a) cholesterol [Lp(a)-C]. The chromatographic method exploits Lp(a)'s unique high content of carbohydrate, e.g. N-acetyl-glucosamine and sialic acid. Lp(a) from serum is separated from other lipoproteins by binding of its carbohydrate structures to a specific solid-supported lectin. The bound Lp(a) is eluted and analyzed by a highly sensitive enzymic cholesterol reagent. The method enables quantitation of Lp(a)-C on clinical analyzers, together with the traditional lipid profile of cholesterol, triglyceride, HDL-C and LDL-C. In the noninstrumented visual teststrip method, blood from a fingerstick is separated into its cellular and plasma components by an integrated separating member. The plasma is transferred into a lectin- or antibody- impregnated Lp(a) recipient member. The bound lp(a) is washed free of other lipoproteins and assayed in situ for Lp(a). PROPOSED COMMERCIAL APPLICATION: Lp(a) is an independent risk factor for atherosclerotic/thrombotic vascular disease. Strong associations of elevated plasma levels have been demonstrated for coronary heart and cerebrovascular disease. In the U.S. about three quarters of a million people die each year from these disorders. The annual socioeconomic toll of heart disease has been estimated at $60 billion. The U.S. reagent market for lipid and lipoprotein testing in excess of $150 million. Laboratory billings are about $1.5 billion. Not a single laboratory method currently exists for the quantitation of the cholesterol component of Lp(a), a potential major contributor to Lp(a) atherogenicity.

据推测，脂蛋白（a）[Lp（a）]可能是最重要的 人类致动脉粥样硬化脂蛋白：脂蛋白（a）的现行实验室方法 采用免疫定量[载脂蛋白（a）]。 因为这些方法阻碍了 通过与纤溶酶原的交叉反应性、载脂蛋白（a）亚型依赖性、抗体 异质性，缺乏标准化，没有一个是FDA批准的。 脂蛋白（a）的三分之一是胆固醇，只有十分之一是载脂蛋白（a）。 我们建议发展一种定量的亲和色谱法， Lp（a）胆固醇[Lp（a）-C]的半定量试纸条法。 的 色谱法利用Lp（a）独特的高含量 碳水化合物，例如N-乙酰基-葡糖胺和唾液酸。 Lp（a）来自 血清通过与其它脂蛋白结合而与其它脂蛋白分离， 碳水化合物结构到特定的固体支持的凝集素。 结合的 脂蛋白（a）是洗脱和分析的高度敏感的酶胆固醇 试剂. 该方法能够定量临床上的Lp（a）-C 分析仪，连同传统的胆固醇血脂谱， 甘油三酯、HDL-C和LDL-C。 在非仪器化的视觉测试条方法中，手指针刺的血液 被集成的分离器分离成细胞和血浆成分 分离元件。 血浆被转移到凝集素-或抗体- 浸渍的Lp（a）受体成员。 将结合的lp（a）洗涤掉， 其他脂蛋白，并原位测定Lp（a）。 拟定商业应用： Lp（a）是动脉粥样硬化/血栓形成的独立危险因素 血管疾病 血浆水平升高与 被证明是冠心病和脑血管病的病因。 在 美国每年大约有75万人死于 紊乱 每年心脏病的社会经济损失 估计为600亿美元。 美国的脂质试剂市场 超过一亿五千万美元的脂蛋白检测 实验室账单是 大约15亿美元。 目前还没有一种实验室方法用于 Lp（a）的胆固醇组分的定量， Lp（a）致动脉粥样硬化性。