DESIGN OF BIOLUMINESCENCE-BASED FIBER OPTIC SENSORS FOR FLOW-INJECTION ANALYSIS

被引:13
作者
BLUM, LJ [1 ]
GAUTIER, SM [1 ]
COULET, PR [1 ]
机构
[1] UNIV LYON 1,CNRS,GENIE ENZYMAT LAB,F-69622 VILLEURBANNE,FRANCE
关键词
FIBER OPTIC; BIOSENSOR; BIOLUMINESCENCE; FLOW INJECTION ANALYSIS; FIA; ATP ANALYSIS; NADH ANALYSIS; ENZYME ACTIVITY ASSAY;
D O I
10.1016/0168-1656(93)90080-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A fiber optic sensor based on enzyme-catalyzed light-emitting reactions has been developed and integrated in a flow-injection analysis (FIA) system. The firefly luciferase, specific for ATP, and the bacterial oxidoreductase/ luciferase system, specific for NADH, have been immobilized on preactivated polyamide membranes. ATP and NADH analysis could be performed in the range from 0.1 pmol to 3 nmol and from 0.5 pmol to 1 nmol, respectively. By co-immobilizing these two bioluminescence systems on the same membrane, a multi-function biosensor has been designed allowing the alternate determination of ATP or NADH with the same sensitivity as that obtained with the two different monofunctional biosensors. A partly self-contained biosensor has been also developed for the flow injection analysis of NADH. For this purpose, FMN (one of the substrates of the bacterial bienzymatic system) has been embedded in a synthetic matrix. Different supports have been tested for the non-covalent immobilization of this substrate and its release in the immediate vicinity of the bound enzymes. Using a photo-crosslinked poly(vinyl alcohol) support, 40 reliable assays (cv = 4.5%) could be performed without changing or reloading the matrix.
引用
收藏
页码:357 / 368
页数:12
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