IMMUNOSENSING WITH AMPEROMETRIC DETECTION, USING GALACTOSIDASE AS LABEL AND P-AMINOPHENYL-BETA-D-GALACTOPYRANOSIDE AS SUBSTRATE

被引：57

作者：

MASSON, M ^{[1
]}

LIU, Z ^{[1
]}

HARUYAMA, T ^{[1
]}

KOBATAKE, E ^{[1
]}

IKARIYAMA, Y ^{[1
]}

AIZAWA, M ^{[1
]}

机构：

[1] TOKYO INST TECHNOL,FAC BIOSCI & BIOTECHNOL,DEPT BIOENGN,MIDORI KU,YOKOHAMA,KANAGAWA 227,JAPAN

来源：

ANALYTICA CHIMICA ACTA | 1995年 / 304卷 / 03期

关键词：

IMMUNOASSAY; AMPEROMETRY; FLOW INJECTION; GALACTOSIDASE; P-AMINOPHENYL-BETA-D-GALACTOPYRANOSIDE;

D O I：

10.1016/0003-2670(94)00645-3

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

p-Aminophenyl-beta-D-galactopyranoside (PAPG) was shown to be a suitable substrate for the amperometric detection of galactosidase activity at neutral pH. The application of this amplification system for immunoassay was demonstrated. The product of the enzyme reaction, p-aminophenol (PAP), was detected at 200 mV, vs. Ag/AgCl, by now-injection analysis (FIA), with a 50 nM detection limit. PAPG was hydrolyzed more than 2.5 times faster than p-nitrophenyl-beta-D-galactopyranoside, by the enzyme, Both PAP and PAPG were stable at pH 7. The galactosidase concentration could be measured down to a concentration of 100 fM, and mouse IgG could be assayed by sandwich immunoassay down to 700 fM. PAPG was found to be a promising reagent for heterogeneous systems, like the one described, and for homogenous assays of biological fluids.

引用

页码：353 / 359

页数：7

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