【乳胶手套洗脱总蛋白的比色法测定】。

T Yagami, M Sato, A Nakamura
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引用次数: 0

摘要

近年来,已报道了由乳胶手套引起的I型超敏反应。乳胶手套中的蛋白质被认为是导致这种过敏的原因。对乳胶手套中总蛋白的提取条件和测定方法进行了研究。一系列实验表明,该蛋白的洗脱速度非常快,提取缓冲液体积(1.7 ~ 20 ml/g)的变化对提取效率没有明显影响。此外,四种测定方法的对比实验表明,Ninhydrin法受蛋白质以外的低分子量物质干扰较大,Bradford法的重现性较差。而Lowry法和BCA法的测定结果相对较好。根据这些实验结果,我们建立了一个典型的总蛋白定量分析程序如下。取2克乳胶手套标本,取10 ml份pH为7.4的磷酸盐缓冲盐水(PBS)在室温下提取2小时,用BCA法测定所得溶液。按此程序对市售的十种乳胶手套进行了分析。我们发现,可提取蛋白质的量与产品有很大的不同。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Colorimetric determination of the total protein eluted from latex gloves].

Recently type I hypersensitivity reactions caused by latex gloves have been reported. The protein from latex gloves is considered to be responsible for this allergy. We investigated the extraction conditions and the assay methods for the purpose of the colorimetric determination of the total protein eluted from latex gloves. In a series of experiments, it was demonstrated that elution of the protein was very rapid, and changes of the extraction buffer volume (1.7-20 ml/g) had no appreciable effect on the extraction efficiency. Moreover, comparison experiments of the four assay methods revealed that Ninhydrin method was significantly interfered by low molecular weight substances other than protein and that Bradford method suffered from the poor reproducibility. On the other hand, Lowry method and Bicinchoninic acid (BCA) method gave relatively good results. Based on these experimental results, we established a typical procedure for quantitative analysis of the total protein as follows. Two grams of latex gloves specimens are extracted with 10 ml portions of phosphate buffered saline pH 7.4 (PBS) at room temperature for two hours, and then the resulting solution is assayed by BCA method. According to this procedure, commercially available ten kinds of latex gloves were analyzed. We found that the amount of the extractable protein was considerably varied with the products.

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