METHOD FOR ISOLATION AND MICROASSAY OF LIPOPROTEIN A

据推测，脂蛋白(A)[Lp(A)]可能是 人体内的致动脉粥样硬化性脂蛋白。脂蛋白(A)的现行实验室检测方法 采用免疫定量[apo(A)]。因为这些方法受到阻碍 通过与纤溶酶原、载脂蛋白(A)亚型依赖、抗体的交叉反应 异质性，缺乏标准化，没有一种是FDA批准的。 Lp(A)的S体重中有三分之一是胆固醇，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层析，并建立一种 Lp(A)胆固醇的半定量测试条法[Lp(A)-C]。这个 利用Lp(A)S独特的高含量的层析方法 碳水化合物，如N-乙酰-氨基葡萄糖和唾液酸。Lp(A)自 血清通过与其他脂蛋白结合而与其他脂蛋白分离 将碳水化合物结构转化为特定的固体支撑凝集素。《边界》 Lp(A)被一种高度敏感的酶胆固醇洗脱和分析 试剂。该方法可用于临床Lp(A)-C的定量 分析仪，与传统的胆固醇血脂谱一起， 甘油三酯、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇 在非仪器视觉测试条方法中，手指上的血液 被分成细胞和血浆成分 分离成员。血浆被转移到凝集素或抗体中- 受孕的LP(A)接受者成员。结合的Lp(A)被洗去 其他脂蛋白，并原位测定Lp(A)。 建议的商业应用： Lp(A)是动脉粥样硬化/血栓形成的独立危险因素 血管疾病。血浆水平升高的强烈关联 已被证实可治疗冠心病和脑血管疾病。在 美国每年约有75万人死于这些疾病 精神错乱。每年心脏病造成的社会经济损失一直是 估计价值600亿美元。美国油脂和食品试剂市场 脂蛋白检测超过1.5亿美元。实验室账单是 大约15亿美元。目前还没有一种实验室方法可以用来 脂蛋白(A)中胆固醇组分的定量 Lp(A)致动脉粥样硬化的因素。

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.