Mulchandani A, Bassi A S
Chemical Engineering Department, University of California, Riverside 92521, USA.
Biosens Bioelectron. 1996;11(3):271-80. doi: 10.1016/0956-5663(96)88414-4.
Tetrathiafulvalene (TTF) mediated amperometric enzyme electrodes have been developed for the monitoring of L-glutamine and L-glutamic acid in growing mammalian cell cultures. The detection of glutamine was accomplished by a coupled enzyme system comprised of glutaminase plus glutamate oxidase, while the detection of glutamic acid was carried out by a single enzyme, glutamate oxidase. The appropriate enzyme(s) were immoblized on the Triton-X treated surface of tetrathiafulvalene modified carbon paste electrodes by adsorption, in conjunction with entrapment by an electrochemically deposited copolymer film of 1,3-phenylenediamine and resorcinol. Operating conditions for the glutamine enzyme electrode were optimized with respect to the amount of enzymes immoblized, pH, temperature and mobile phase flow rate for operation in a flow injection (FIA) system. When applied to glutamine and glutamic acid measurements in mammalian cell culture in FIA, the results obtained with enzyme electrodes were in excellent agreement with those determined by enzymatic analysis.
已开发出四硫富瓦烯（TTF）介导的安培型酶电极，用于监测生长中的哺乳动物细胞培养物中的L-谷氨酰胺和L-谷氨酸。谷氨酰胺的检测通过由谷氨酰胺酶加谷氨酸氧化酶组成的偶联酶系统完成，而谷氨酸的检测则由单一酶谷氨酸氧化酶进行。通过吸附将适当的酶固定在四硫富瓦烯修饰的碳糊电极经曲拉通-X处理的表面上，并与1,3-苯二胺和间苯二酚的电化学沉积共聚物膜包埋相结合。针对固定化酶的量、pH、温度和流动注射（FIA）系统中操作的流动相流速，对谷氨酰胺酶电极的操作条件进行了优化。当将酶电极应用于FIA中哺乳动物细胞培养物中谷氨酰胺和谷氨酸的测量时，酶电极获得的结果与酶分析确定的结果非常一致。
