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: 2029566
• 负责人: Mary Gaunt-Kloepfer
• 金额: $ 30.87万
• 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 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2029566
• 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。 人类的致动脉粥样硬化脂蛋白。 目前 Lp(a) 的实验室方法 采用免疫定量[apo(a)]。 因为这些方法都受到阻碍 通过与纤溶酶原的交叉反应、apo(a) 同工型依赖性、抗体 异质性和缺乏标准化，它们都没有获得 FDA 批准。 Lp(a) 质量的三分之一是胆固醇，只有十分之一是apo (a)。 我们建议开发一种定量亲和色谱法，以及 Lp(a) 胆固醇的半定量测试条方法 [Lp(a)-C]。 这 色谱法利用了 Lp(a) 独特的高含量 碳水化合物，例如N-乙酰氨基葡萄糖和唾液酸。 Lp(a) 来自 血清通过结合其与其他脂蛋白分离 将碳水化合物结构转化为特定的固体支持的凝集素。 界限 Lp(a) 通过高度灵敏的酶胆固醇洗脱并分析 试剂。 该方法能够在临床上定量 Lp(a)-C 分析仪以及传统的胆固醇脂质谱， 甘油三酯、HDL-C 和 LDL-C。 在非仪器视觉试纸法中，指尖采血 通过集成的系统将其分离成细胞和血浆成分 分离成员。 血浆被转移到凝集素或抗体中 浸渍的 Lp(a) 受体成员。 结合的 lp(a) 被洗掉 其他脂蛋白并原位测定 Lp(a)。 拟议的商业应用： Lp(a)是动脉粥样硬化/血栓形成的独立危险因素 血管疾病。 血浆水平升高有很强的关联性 已被证明可治疗冠心病和脑血管疾病。 在 美国每年约有四分之三的人死于这些疾病 失调。 心脏病每年造成的社会经济损失为 估计为600亿美元。 美国脂质和试剂市场 脂蛋白检测超过 1.5 亿美元。 实验室账单是 约15亿美元。 目前尚不存在单一的实验室方法 Lp(a) 中胆固醇成分的定量，这是一种潜在的主要成分 Lp(a) 致动脉粥样硬化的贡献者。