Sulter M W, Kloosterhuis G J, Coenraads P J, Pas H H
Department of Dermatology, State University Hospital, Groningen, The Netherlands.
Anal Biochem. 1993 Jun;211(2):301-4. doi: 10.1006/abio.1993.1273.
A solid-phase protein assay (SPA) for the determination of protein concentrations in the nanogram range is described. The technique is based on biotinylation of immobilized protein on the solid phase of a microtiter plate and quantitation of the protein-biotin complexes by peroxidase-coupled avidin. Compared to the routinely used Bradford and Lowry protein detection techniques, the described SPA assay is (depending upon the protein assayed) 1000 to 10,000 times more sensitive. Moreover, the SPA assay can be suitable for protein quantitation of samples containing agents which commonly interfere with the routinely used detection techniques. The SPA assay is a valuable addition to the Bradford and the Lowry techniques. Used in combination with an antigen-specific ELISA it gives a reliable ratio of specific versus total protein.
本文描述了一种用于测定纳克级蛋白质浓度的固相蛋白质分析（SPA）方法。该技术基于对微量滴定板固相上固定化蛋白质进行生物素化，并通过过氧化物酶偶联抗生物素蛋白对蛋白质-生物素复合物进行定量。与常规使用的考马斯亮蓝法和洛瑞蛋白质检测技术相比，所述的SPA分析方法（取决于所检测的蛋白质）灵敏度高1000至10000倍。此外，SPA分析方法适用于对含有通常会干扰常规检测技术的试剂的样品进行蛋白质定量。SPA分析方法是对考马斯亮蓝法和洛瑞技术的有价值补充。与抗原特异性酶联免疫吸附测定（ELISA）结合使用时，它能给出特异性蛋白质与总蛋白质的可靠比例。
