用于测定浑浊生物流体中特定蛋白质的激光散射比浊法等效点滴定法。

Laser nephelometric equivalence point titration for the measurements of specific proteins in turbid biological fluids.

作者信息

Hobler H, Schütterle U, Nickel S

出版信息

J Clin Chem Clin Biochem. 1985 Feb;23(2):89-97. doi: 10.1515/cclm.1985.23.2.89.

DOI:10.1515/cclm.1985.23.2.89

Abstract

A reliable, sensitive and specific laser nephelometric (LN) assay for the quantitative measurement of specific proteins in body fluids by equivalence point titration is described. The method is relatively free from interference encountered in conventional LN assays, e.g. unspecific background scatter, antigen excess, and the inhibition of complex formation by high ionic strength and salt concentration, low pH and elevated urea concentrations. It can be applied to turbid body fluids containing relatively small amounts of antigen. A serial dilution of the antigen preparation (human urinary albumin in the present work) is incubated with a fixed amount of highly diluted specific antiserum. The antigen concentration is determined by estimation of the equivalence point on the LN immunoprecipitation curve and comparison of this position with that on a standard curve. The data presented show good correlation with values obtained by radioimmunoassay (p less than or equal to 1 X 10(-5);log (LNET) = 0.956 log (RIA) - 0.03; n = 50).

摘要

本文描述了一种可靠、灵敏且特异的激光散射比浊法（LN），通过等价点滴定法对体液中的特定蛋白质进行定量测定。该方法相对不受传统LN测定中遇到的干扰影响，例如非特异性背景散射、抗原过量以及高离子强度、盐浓度、低pH值和高尿素浓度对复合物形成的抑制。它可应用于含有相对少量抗原的浑浊体液。将抗原制剂（本研究中为人尿白蛋白）的系列稀释液与固定量的高度稀释的特异性抗血清孵育。通过估计LN免疫沉淀曲线上的等价点，并将该位置与标准曲线上的位置进行比较来确定抗原浓度。给出的数据与放射免疫测定获得的值具有良好的相关性（p≤1×10⁻⁵；log（LNET）=0.956 log（RIA） - 0.03；n = 50）。