A comparison of silver ion to streptavidin coated microplates.
Direct comparisons are made between covalently linked streptavidin and silver ion coated microplates. Both coatings can immobilize biotinylated molecules. Silver ion coated microplate wells can immobilize 1.8 times higher amounts of biotin labeled horseradish peroxidase. The quantitation range and capacity for the capture of horseradish peroxidase using biotin labeled horseradish peroxidase are also greater for silver ion coated microplates. Approximately twice as many anti-horseradish peroxidase antibodies can be immobilized per well using silver ion coated microplates. Higher capacities are presumed to be due to the smaller footprint of silver ions as compared to streptavidin. A direct comparison between the two coatings for a beta-galactosidase ELISA showed that while the silver ion coated microplates gave higher readings, the streptavidin coated microplates exhibited smaller well-to-well variation. However, higher well to well variation for the silver microplates is attributed to the high density of anti-beta-galactosidase antibodies on the microplates and the weak binding of clone GAL-13 to beta-galactosidase, rather than the silver coating. These studies suggest silver ion coated microplates are a desirable alternative to streptavidin plates for quantitative immunoassays.
Bonen
M R
MR
Department of Chemical and Materials Engineering, Arizona State University, Tempe, AZ 85287-6006, USA.
García
A A
AA
Hoffman
S A
SA
D000911:Antibodies, Monoclonal / Q000032:analysis; D001710:Biotin / Q000378:metabolism*; D019921:Biotinylation / Q000295:instrumentation* / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006735:Horseradish Peroxidase / Q000032:analysis / Q000276:immunology; D012834:Silver; D019809:Streptavidin; D001616:beta-Galactosidase / Q000378:metabolism
D000911:Antibodies, Monoclonal; D012834:Silver; D001710:Biotin; D019809:Streptavidin; D006735:Horseradish Peroxidase; D001616:beta-Galactosidase