Maebashi Institute of Technology, 371-0816 Maebashi, Gunma, Japan.
Maebashi Institute of Technology, 371-0816 Maebashi, Gunma, Japan.
Anal Chim Acta. 2014 Mar 3;814:55-62. doi: 10.1016/j.aca.2014.01.031. Epub 2014 Jan 19.
To electrochemically detect concanavalin A (ConA), a new method was developed using mixed micelles between a non-ionic surfactant with a maltose moiety and electroactive daunomycin. The surfactants, in which the length of the alkyl chain was different, were n-decyl-β-D-maltoside, n-dodecyl-β-D-maltoside, and n-tetradecyl-β-D-maltoside. The measurement principle was due to the micelle breakdown caused by the binding between the ConA and maltose moieties. When ConA was combined with maltose moieties at a concentration of surfactant that was near the critical micelle concentration, the daunomycin that formed the micelles was moved to a solution from the micelles. As a result, the peak current of daunomycin increased as the concentration of ConA was increased. The mechanism was proposed using voltammetry, spectrometry, and gel filtration. The linear range using n-tetradecyl-β-D-maltoside was 2.0×10(-9) to 8.0×10(-8) M of ConA, and it was the most sensitive in the presence of the three surfactants. To examine whether selective binding took place, measurements with several proteins were carried out. The electrode responses of daunomycin were not influenced by the presence of 5.0×10(-6) M protein. Furthermore, this method could be applied to the determination of ConA in a serum, and to the measurement of sugar chains that can be combined with ConA on the cell surface.
为了电化学检测伴刀豆球蛋白 A(ConA),开发了一种新方法,该方法使用带有麦芽糖部分的非离子表面活性剂与具有电化学活性的柔红霉素的混合胶束。表面活性剂的烷基链长度不同,分别为正癸基-β-D-麦芽糖苷、正十二烷基-β-D-麦芽糖苷和正十四烷基-β-D-麦芽糖苷。该测量原理是由于 ConA 与麦芽糖部分之间的结合导致胶束破裂。当 ConA 与麦芽糖部分结合时,其浓度接近临界胶束浓度,形成胶束的柔红霉素从胶束转移到溶液中。结果,随着 ConA 浓度的增加,柔红霉素的峰电流增加。该机制是通过伏安法、光谱法和凝胶过滤法提出的。使用正十四烷基-β-D-麦芽糖苷的线性范围为 2.0×10(-9)至 8.0×10(-8) M 的 ConA,在三种表面活性剂中最灵敏。为了检查是否发生了选择性结合,对几种蛋白质进行了测量。存在 5.0×10(-6) M 蛋白质时,柔红霉素的电极响应不受影响。此外,该方法可用于测定血清中的 ConA,以及测量可与细胞表面上的 ConA 结合的糖链。