Scaled down method for the reproducible recovery to high purity of human serum albumin from low volume blood samples.

Bioseparation Pub Date : 2001-01-01 DOI:10.1023/a:1016347102660

M Rynö, T Anttinen-Klemetti, R Vaaranrinta, J Tornaeus, A Hesso, T Veidebaum, K Peltonen

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引用次数: 1

Abstract

A basic need for a protein-based dosimeter is a purified protein. In this communication we present an isolation protocol and an HPLC-based assay which allows one to determine the purity of the isolated albumin. A total of 168 human blood samples were collected from workers of a benzene processing plant and from nearby countryside at Kohtla-Järve, Estonia. Albumin was isolated from plasma by sequential precipitation and the purity was determined by HPLC. The amount of albumin present in plasma varied between the individuals, being 147 +/- 26 mg/5 ml (n = 168), which is about 59% of plasma albumin. However, the isolated albumin was highly pure (100.9 +/- 8.2%, n = 5). All albumin samples analyzed demonstrate two peaks in HPLC analysis. The two peaks detected were collected and subjected to MS analysis, which demonstrates a difference of 120 mass units between the two albumin products isolated. We have developed an assay, which is easy to carry out and is not too labor intense. The HPLC analysis can be applied to confirm the purity of the isolated albumin as well as to confirm the quantity of the albumin in samples.

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从小容量血液样品中可重复回收高纯度人血清白蛋白的缩小方法。

蛋白质剂量计的基本需求是纯化的蛋白质。在本通讯中，我们提出了一种分离方案和一种基于高效液相色谱的测定方法，可以确定分离白蛋白的纯度。从爱沙尼亚Kohtla-Järve的苯加工厂和附近农村的工人共收集了168份人体血液样本。用顺序沉淀法从血浆中分离白蛋白，用高效液相色谱法测定其纯度。血浆中白蛋白的含量因人而异，为147 +/- 26 mg/5 ml (n = 168)，约占血浆白蛋白的59%。然而，分离的白蛋白纯度很高(100.9 +/- 8.2%，n = 5)。所有白蛋白样品在HPLC分析中都显示出两个峰。将检测到的两个峰收集并进行质谱分析，结果表明分离的两种白蛋白产物之间存在120个质量单位的差异。我们开发了一种易于操作且劳动强度不高的化验方法。高效液相色谱分析可用于确定分离白蛋白的纯度和样品中白蛋白的数量。

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Bioseparation

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