Reduced immunoreactivity of urinary albumin in patients with cardiovascular diseases: Analysis of immunochemically nonreactive albumin

Journal of capillary electrophoresis Pub Date : 2008-09-01 DOI:10.2198/JELECTROPH.52.57

Akinobu Nakayama, Jyunichi Nishimaki, T. Kawara, T. Kasama, Toshiaki Baba, H. Yoshida, M. Isobe, K. Shiba, Kenji Sato

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引用次数: 1

Abstract

We analyzed 55 spot urine samples from patients with cardiovascular diseases. Urinary albumin concentrations were measured with size exclusion high performance liquid chromatography (HPLC), turbidimetric immunoassay (TIA) using anti human serum albumin polyclonal antibody, and enzyme-linked immunosorbent assay (ELISA) using anti human serum albumin monoclonal antibody. Fractionated urine samples from the HPLC were also analyzed with the immunoprecipitation reactions using as same monoclonal antibody as ELISA. As a result, the urinary albumin concentration analyzed by the HPLC was systematically higher than that of immunoassays, however, ‘albumin peak’ from the HPLC contained other urinary proteins. Result of immunoprecipitation reaction showed the presence of monomer albumin that could not react with the monoclonal antibody. These results suggest that not only contamination of other proteins, the albumin fraction from the HPLC included albumin with reduced its reactivity to the specific monoclonal antibody.

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心血管疾病患者尿白蛋白免疫反应性降低:免疫化学非反应性白蛋白的分析

我们分析了55例心血管疾病患者的尿样。尿白蛋白浓度测定采用粒径排除高效液相色谱法(HPLC)，浊度免疫法(TIA)采用抗人血清白蛋白多克隆抗体，酶联免疫吸附法(ELISA)采用抗人血清白蛋白单克隆抗体。HPLC分离的尿样也使用与ELISA相同的单克隆抗体进行免疫沉淀反应。结果，HPLC分析的尿白蛋白浓度系统地高于免疫分析法，然而，HPLC的“白蛋白峰”含有其他尿蛋白。免疫沉淀反应结果显示存在不能与单克隆抗体发生反应的单体白蛋白。这些结果表明，不仅其他蛋白质受到了污染，HPLC检测的白蛋白部分还含有白蛋白，其对特异性单克隆抗体的反应性降低。

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Journal of capillary electrophoresis

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