Riechel P, Weiss T, Weiss M, Ulber R, Buchholz H, Scheper T
Institut für Technische Chemie, Universität Hannover, Germany.
J Chromatogr A. 1998 Aug 21;817(1-2):187-93. doi: 10.1016/s0021-9673(98)00445-2.
In our present work we present the determination of bovine lactoferrin in whey concentrates as they are typically produced by milk and cheese industry after production of cheese. Due to the high total protein content the analysis of whey concentrate samples is difficult and even not possible by using capillary zone electrophoresis with UV detection. To enhance the detection sensitivity we applied a more promising approach by using affinity interactions in combination with laser-induced fluorescence detection. By mixing fluoresceine isothiocyanate (FITC)-conjugated polyanionic lipopolysaccharide with the mostly positively charged lactoferrin we found a significant migration time shift which is clearly dependent on the concentration of the added protein. In the second approach we developed an immunoassay using FITC-conjugated specific antibody against bovine lactoferrin. The results of the immunoassay measurements were compared with data obtained by standard enzyme-linked immunosorbent assay analysis.
在我们目前的工作中，我们展示了对乳清浓缩物中牛乳铁蛋白的测定，这些乳清浓缩物通常是在奶酪生产后由牛奶和奶酪行业生产的。由于总蛋白含量高，使用紫外检测的毛细管区带电泳分析乳清浓缩物样品很困难，甚至不可能。为了提高检测灵敏度，我们采用了一种更有前景的方法，即利用亲和相互作用结合激光诱导荧光检测。通过将异硫氰酸荧光素（FITC）偶联的聚阴离子脂多糖与大多带正电荷的乳铁蛋白混合，我们发现迁移时间有显著变化，这明显取决于添加蛋白质的浓度。在第二种方法中，我们开发了一种使用针对牛乳铁蛋白的FITC偶联特异性抗体的免疫测定法。将免疫测定测量结果与通过标准酶联免疫吸附测定分析获得的数据进行了比较。
