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髓磷脂碱性蛋白的放射免疫测定。方法学评价。

Radioimmunoassay of myelin basic protein. A methodological evaluation.

作者信息

Karlsson B, Alling C

出版信息

J Immunol Methods. 1982 Nov 26;55(1):51-61. doi: 10.1016/0022-1759(82)90076-x.

Abstract

Three techniques for separating free antigen from antigen-antibody complexes have been applied to radioimmunoassay of myelin basic protein: cold ethanol precipitation of complexes, dextran-coated charcoal precipitation of free antigen, and second antibody precipitation of complexes. After optimization of the incubation and separation steps, the 3 methods were evaluated for precision and accuracy when applied to both spinal fluid and brain tissue homogenates. For determinations in brain tissue all 3 methods showed the same precision and gave largely the same values, though the ethanol method gave slightly lower levels. For spinal fluid the ethanol and dextran-charcoal methods gave the same values, but the double antibody method gave values only 1/3 as high. With spinal fluid, the precision of the dextran-charcoal method was poor compared with that of the other two. The double antibody method proved to be the method of choice for brain tissue samples, when the results of the incubation and separation steps, and the precision and accuracy of the determinations were taken into account. However, for an unknown reason values for spinal fluid were too low by this method. Therefore the ethanol precipitation method is recommended for spinal fluid samples and the double antibody method for brain tissue samples.

摘要

三种从抗原-抗体复合物中分离游离抗原的技术已应用于髓鞘碱性蛋白的放射免疫分析:复合物的冷乙醇沉淀、游离抗原的葡聚糖包被活性炭沉淀以及复合物的第二抗体沉淀。在优化孵育和分离步骤后,对这三种方法应用于脑脊液和脑组织匀浆时的精密度和准确性进行了评估。对于脑组织中的测定,所有三种方法均显示出相同的精密度,并且给出的值大致相同,尽管乙醇法得到的值略低。对于脑脊液,乙醇法和葡聚糖-活性炭法给出的值相同,但双抗体法给出的值仅为其三分之一。对于脑脊液,与其他两种方法相比,葡聚糖-活性炭法的精密度较差。考虑到孵育和分离步骤的结果以及测定的精密度和准确性,双抗体法被证明是脑组织样本的首选方法。然而,由于未知原因,该方法测定的脑脊液值过低。因此,推荐乙醇沉淀法用于脑脊液样本,双抗体法用于脑组织样本。

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