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岩藻依聚糖特异性抗体的研制及采用夹心酶联免疫吸附测定法检测血清和尿液中的岩藻依聚糖

Development of a fucoidan-specific antibody and measurement of fucoidan in serum and urine by sandwich ELISA.

作者信息

Tokita Yoshiharu, Nakajima Katsuyuki, Mochida Hiroshi, Iha Masahiko, Nagamine Takeaki

机构信息

School of Health Sciences, Faculty of Medicine, Gunma University, Maebashi, Japan.

出版信息

Biosci Biotechnol Biochem. 2010;74(2):350-7. doi: 10.1271/bbb.90705. Epub 2010 Feb 7.

Abstract

Fucoidan exhibits various biological properties. We raised a novel antibody against fucoidan extracted from Cladosiphon okamuranus and developed a sandwich ELISA method to measure fucoidan. The fucoidan antibody was specific and did not cross-react with other polysulfated polysaccharides. Fucoidan recovery from serum and urine by ELISA was 86-113%. Intra- and inter-assay CVs were 1.5-13.4%. Assay linearity was maintained after 3-fold dilution of each sample with phosphate-buffer saline (PBS). In the serum and urine of healthy volunteers (n=10), fucoidan was not detected before administration, and the levels markedly increased 6 and 9 h after oral administration. The molecular weight of the serum fucoidan determined by HPLC gel filtration remained unchanged, whereas that of urine fucoidan was significantly reduced. This is the first ELISA method of measuring serum and urine fucoidan levels after oral administration. The method is simple, reliable, and practical for the analysis of samples, especially urine samples.

摘要

岩藻依聚糖具有多种生物学特性。我们制备了一种针对从冈村枝管藻中提取的岩藻依聚糖的新型抗体,并开发了一种夹心酶联免疫吸附测定(ELISA)法来检测岩藻依聚糖。该岩藻依聚糖抗体具有特异性,不会与其他多硫酸化多糖发生交叉反应。通过ELISA从血清和尿液中回收岩藻依聚糖的回收率为86%-113%。批内和批间变异系数为1.5%-13.4%。用磷酸盐缓冲盐水(PBS)对每个样品进行3倍稀释后,测定线性得以维持。在健康志愿者(n=10)的血清和尿液中,口服给药前未检测到岩藻依聚糖,口服给药后6小时和9小时其水平显著升高。通过高效液相色谱(HPLC)凝胶过滤法测定的血清岩藻依聚糖的分子量保持不变,而尿液岩藻依聚糖的分子量则显著降低。这是第一种用于测定口服给药后血清和尿液中岩藻依聚糖水平的ELISA方法。该方法对于样品尤其是尿液样品的分析而言简单、可靠且实用。

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