RADIOIMMUNOASSAY FOR HUMAN LIPOPROTEIN LIPASE

该研究项目的目的是开发和验证一种精确的， 脂蛋白脂酶非竞争固相放射免疫分析 (LPL)肝素前和肝素后的人血浆。 该方法已被 在我们的实验室里为鸟类LPL开发的。 为了建立一个类似的实验 对于人LPL，必须将牛乳LPL纯化至均匀。 针对这种酶的抗血清将在山羊和免疫球蛋白中产生 在LPL-Sepharose亲和柱上纯化。 免疫球蛋白将与 涉及亲水性聚丙烯酰胺珠。 这些免疫球将用于 与125 I免疫球蛋白结合， 放射免疫法 将评价试验的精密度和准确度， 以及血浆蛋白和外源性化学物质的可能干扰， 肝素 肝素治疗前和治疗后正常人和贫血人中的LPL活性 将确定血浆。 一种类似的放射免疫测定法将被开发出来 使用人LPL和抗LPL免疫球蛋白。 的理化 将高度纯化的人LPL的性质与肝LPL的性质进行比较， 脂肪酶。 这一假说认为，连续的结合和释放丰富的甘草酸， 脂蛋白与血管内皮细胞的结合导致血管内皮细胞的逐渐富集， 将测试脂蛋白与LPL蛋白的结合。 LPL和VLDL的摩尔比 还将测定通过速率分区超离心分离的亚组分。

The purpose of this research project is to develop and validate a precise and sensitive non-competitive solid phase radioimmunoassay for lipoprotein lipase (LPL) for human pre- and post-heparin plasma. The method has already been developed in our laboratory for avian LPL. In order to set up a similar assay for human LPL, it will be necessary to purify bovine milk LPL to homogeneity. Antisera against this enzyme will be produced in goats and immunoglobulins purified on LPL-Sepharose affinity columns. The immunoglobulins will be coupled to hydrophilic polyacrylamide beads. These immunobeads will be employed in conjunction with 125I immunoglobulins in a direct sandwich-type radioimmunoassay. The precision and accuracy of the assay will be evaluated, as well as possible interference by plasma proteins and exogenous chemicals such as heparin. LPL activities in normal and hyperlipemic pre- and post-heparin human plasma will be determined. A similar radioimmunoassay will be developed employing human LPL and anti-LPL immunoglobulins. The physical and chemical properties of highly purified human LPL will be compared to those of liver lipase. The hypothesis that successive binding and release of triglyceride-rich lipoproteins with the vascular endothelium leads to a gradual enrichment of the lipoprotein with LPL proteins will be tested. The molar ratios of LPL and VLDL subfractions resolved by rate zonal ultracentrifugation will also be determined.