Evaluation of β-D-Galactosidase from Escherichia Coli and Horseradish Peroxidase as Labels by Sandwich Enzyme Immunoassay Technique
Evaluation of β-D-Galactosidase from Escherichia Coli and Horseradish Peroxidase as Labels by Sandwich Enzyme Immunoassay Technique
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三明治酶联免疫分析技术评价大肠杆菌β-D-半乳糖苷酶和辣根过氧化物酶的标记
DOI：
10.1177/000456328402100414
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发表时间：
1984
影响因子：
2.2
通讯作者：
E. Ishikawa
中科院分区：
文献类型：
--
作者：
M. Imagawa;S. Hashida;Y. Ohta;E. Ishikawa
以荧光底物为酶标，采用夹心酶免疫分析技术，对大肠杆菌β-D-半乳糖苷酶和辣根过氧化物酶作为Fab'的标记物，在人甲胎蛋白和人绒毛膜促性腺激素的量效曲线上进行了评价。Fab ′-过氧化物酶偶联物与IgG包被的聚苯乙烯球的非特异性结合小于Fab ′-β-d-半乳糖苷酶偶联物，亲和纯化的Fab ′-过氧化物酶偶联物对这些抗原给出了比相应的β-d-半乳糖苷酶偶联物更灵敏的剂量-反应曲线。然而，需要大量的Fab '-过氧化物酶缀合物，并且过氧化物酶测定需要更长的孵育时间，因为过氧化物酶测定的灵敏度远低于β-d-半乳糖苷酶测定。对这两种酶的其他优缺点进行了讨论。
β-d-galactosidase from Escherichia coli and horseradish peroxidase were evaluated as labels of Fab' in dose-response curves for human α-fetoprotein and human chorionic gonadotropin by sandwich enzyme immunoassay technique using fluorogenic substrates for enzyme assay. The non-specific binding of Fab'–peroxidase conjugates to IgG-coated polystyrene balls was less than that of Fab'-β-d-galactosidase conjugates, and the affinity-purified Fab'–peroxidase conjugates gave more sensitive dose-response curves for these antigens than the corresponding β-d-galactosidase conjugates. However, a large quantity of Fab'–peroxidase conjugates was required and a longer incubation was necessary for the peroxidase assay, since the peroxidase assay was much less sensitive than the β-d-galactosidase assay. Other advantages and disadvantages of the two enzymes are discussed.