DEVELOPMENT OF A HETEROGENEOUS AMPEROMETRIC IMMUNOSENSOR FOR THE DETERMINATION OF APOLIPOPROTEIN-E IN SERUM

被引:28
作者
MEUSEL, M
RENNEBERG, R
SPENER, F
SCHMITZ, G
机构
[1] INST CHEM & BIOCHEM SENSOR RES,D-48149 MUNSTER,GERMANY
[2] UNIV MUNSTER,DEPT BIOCHEM,D-48149 MUNSTER,GERMANY
[3] UNIV REGENSBURG,INST CLIN CHEM & LAB MED,D-93053 REGENSBURG,GERMANY
关键词
ALKALINE PHOSPHATASE; AMPEROMETRIC IMMUNOSENSOR; APOLIPOPROTEIN E; CARBON ELECTRODE; GLUCOSE OXIDASE; P-AMINOPHENYL PHOSPHATE; SITE-DIRECTED IMMOBILIZATION;
D O I
10.1016/0956-5663(95)96933-P
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Apolipoprotein (ape) E is a constituent of serum lipoproteins and can serve as a diagnostic parameter for the assessment of disorders of lipid metabolism. For the determination of apo E in serum a sandwich-type amperometric immunosensor using disposable membranes was developed. The best results were obtained by using a site-directed attachment of a monoclonal capture antibody to a hydrazide-functionalized membrane surface. Bound antigen was then determined with the aid of a polyclonal antibody labelled with alkaline phosphatase in conjunction with p-aminophenyl phosphate as substrate. In this approach a carbon working electrode (vs. Hg/HgCl2) was used, and enzymatically generated p-aminophenol could be monitored with a detection limit of 40 pmol and a linear range of 26-20000 nM. The sensor displayed a linear response from 50 to 1000 ng ml(-1) apo E. In contrast, antibody coupling through primary amino groups led to a total loss of antigen binding capacity in this assay configuration. The approach with site-directed immobilization however, not only allowed the determination of apo E in serum, but also the determination of antibody cross-reactivity against the apolipoproteins AI, AII and B.
引用
收藏
页码:577 / 586
页数:10
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