Evaluation of beta-D-galactosidase from Escherichia coli and horseradish peroxidase as labels by sandwich enzyme immunoassay technique

beta-D-galactosidase from Escherichia coli and horseradish peroxidase were evaluated as labels of Fab' in dose-response curves for human alpha-fetoprotein and human chorionic gonadotropin by sandwich enzyme immunoassay technique using fluorogenic substrates for enzyme assay. The non-specific bi...

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Bibliographic Details
Published inAnnals of clinical biochemistry Vol. 21 ( Pt 4); p. 310
Main Authors Imagawa, M, Hashida, S, Ohta, Y, Ishikawa, E
Format Journal Article
LanguageEnglish
Published England 01.07.1984
Subjects
alpha-Fetoproteins - immunology
Antigen-Antibody Reactions
beta-Galactosidase
Chorionic Gonadotropin - immunology
Escherichia coli - enzymology
Galactosidases
Horseradish Peroxidase
Humans
Immunoenzyme Techniques
Immunoglobulin Fab Fragments - analysis
Microspheres
Peroxidases
Polystyrenes
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Summary:beta-D-galactosidase from Escherichia coli and horseradish peroxidase were evaluated as labels of Fab' in dose-response curves for human alpha-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'-beta-D-galactosidase conjugates, and the affinity-purified Fab'-peroxidase conjugates gave more sensitive dose-response curves for these antigens than the corresponding beta-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 beta-D-galactosidase assay. Other advantages and disadvantages of the two enzymes are discussed.
ISSN:0004-5632
DOI:10.1177/000456328402100414