Abu-Rabeah Khalil, Polyak Boris, Ionescu Rodica E, Cosnier Serge, Marks Robert S
The National Institute for Biotechnology in the Negev and Department of Biotechnology Engineering, Ben-Gurion University of the Negev, P.O. Box 653, Beer Sheva 84105, Israel.
Biomacromolecules. 2005 Nov-Dec;6(6):3313-8. doi: 10.1021/bm050339j.
N-(3-Aminopropyl)pyrrole was covalently coupled with alginate in an aqueous-phase reaction by means of carbodiimide-mediated activation chemistry to provide a pyrrole-alginate conjugate for subsequent use in biosensor applications. The pyrrole-alginate conjugate was quantified by UV spectroscopy at 230 nm, by an HPSEC-MALLS analytical method, as well as by FTIR and 13C NMR spectroscopies. The new pyrrole-alginate conjugate was used for the immobilization of polyphenol oxidase (PPO) onto an electrode surface by physical entrapment resulting from the gellification process and electrochemical polymerization of the pyrrole groups. The efficiency of this cross-linking approach (chemical and electrochemical) was investigated by comparing the amount of enzyme released from polypyrrole-alginate and regular alginate. In addition, biosensors were prepared by entrapment of the PPO in polypyrrole-alginate and regular alginate matrixes and their performance for the amperometric determination of catechol chosen as a model analyte was examined, yielding a sensitivity of 350 and 80 microA M(-1) cm(-2), respectively, for polypyrrole-alginate and alginate biosensors.
通过碳二亚胺介导的活化化学方法,在水相反应中将N-(3-氨丙基)吡咯与海藻酸盐共价偶联,以提供一种吡咯-海藻酸盐共轭物,用于后续的生物传感器应用。通过紫外光谱在230nm处、高效体积排阻色谱-多角度激光光散射(HPSEC-MALLS)分析方法以及傅里叶变换红外光谱(FTIR)和碳-13核磁共振光谱(13C NMR)对吡咯-海藻酸盐共轭物进行定量。新的吡咯-海藻酸盐共轭物用于通过凝胶化过程和吡咯基团的电化学聚合导致的物理包埋将多酚氧化酶(PPO)固定在电极表面。通过比较从聚吡咯-海藻酸盐和普通海藻酸盐中释放的酶量,研究了这种交联方法(化学和电化学)的效率。此外,通过将PPO包埋在聚吡咯-海藻酸盐和普通海藻酸盐基质中来制备生物传感器,并检测它们对作为模型分析物的儿茶酚进行安培测定时的性能,聚吡咯-海藻酸盐生物传感器和海藻酸盐生物传感器的灵敏度分别为350和80μA M(-1) cm(-2)。