Institute of Analytical Chemistry Academy of Sciences of the Czech Republic, v. v. i., 602 00 Brno, Czech Republic.
Anal Chem. 2009 Aug 15;81(16):6897-904. doi: 10.1021/ac900897c.
A chromophoric nonionogenic surfactant poly(ethylene glycol) 3-(2-hydroxy-5-n-octylphenylazo)-benzoate, HOPAB, has been prepared and used as a buffer additive for a dynamic modification of proteins and/or microorganisms including Escherichia coli , Staphylococcus epidermidis (biofilm-positive and biofilm-negative), and the strains of yeast cells Candida albicans and Candida parapsilosis (biofilm-positive and biofilm-negative) during a capillary electrophoresis and a capillary isoelectric focusing (CIEF) with UV detection at 326 nm. Values of isoelectric points of labeled proteins and microorganisms have been calculated using UV-detectable pI markers and have been found comparable with pI of the native compounds. Minimum detectable amount has been assessed lower than picograms of proteins and lower than a hundred cells injected into a separation capillary. The introduced labeling method facilitates CIEF separation of microorganisms from the clinical sample of the infected urine at their clinically important levels in the pH gradient pH range of 2-5 and their subsequent cultivation. At the same time, it has enabled the determination of albumin in human urine as a major clinical marker of urinary tract infections and kidney diseases.
一种发色非离子型表面活性剂聚乙二醇 3-(2-羟基-5-正辛基偶氮苯)-苯甲酸酯（HOPAB）已被制备并用作缓冲添加剂，用于对蛋白质和/或微生物（包括大肠杆菌、表皮葡萄球菌（生物膜阳性和生物膜阴性）和酵母细胞的菌株白色念珠菌和近平滑念珠菌（生物膜阳性和生物膜阴性））进行动态修饰，在毛细管电泳和毛细管等电聚焦（CIEF）中，在 326nm 处进行 UV 检测。使用可检测 UV 的 pI 标记物计算了标记蛋白质和微生物的等电点值，并发现与天然化合物的 pI 相当。最低可检测量评估为低于皮克级的蛋白质和低于注入分离毛细管的一百个细胞。引入的标记方法促进了在 pH 梯度 2-5 范围内从感染尿液的临床样本中分离微生物，以及在该 pH 梯度范围内进行分离后对其进行培养。同时，它还能够测定人尿液中的白蛋白，白蛋白是人尿液中尿路感染和肾脏疾病的主要临床标志物。
