A Small Scale Sandwich Enzyme Immunoassay for Macromolecular Antigens Using B-D-Galactosidase from Escherichia Coli and Horseradish Peroxidase as Labels
- 1 April 1987
- journal article
- research article
- Published by Taylor & Francis in Analytical Letters
- Vol. 20 (4), 587-601
- https://doi.org/10.1080/00032718708067991
Abstract
A small scale sandwich enzyme immunoassay for macromolecular antigens using β-D-galactosidase from Escherichia coli and horseradish peroxidase as labels is described. An antibody IgG-coated glass ball of 1.0 mm in diameter was incubated with antigen in 5 μl and subsequently with affinity-purified Fab'-enzyme conjugate in 5 μl. Using B-D-galactosidase, the detection limits of ferritin, IgE, a-fetoprotein and chorionic gonadotropin were 0.001 amol (600 molecules), 0.03 amol, 0.03 amol and 0.07 amol, respectively. Using peroxidase, the detection limits of ferritin, α-fetoprotein and chorionic gonadotropin were 0.005 amol, 0.1 amol and 0.2 amol, respectively. These detection limits were 3 to 30-fold less than those obtained by the previous sandwich enzyme imnunoassays using antibody IgG-coated polystyrene balls of 3.2 mn in diameter in 150 μl.Keywords
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