A silicon sensor-based filtration immunoassay using biotin-mediated capture.
A sensitive sandwich immunoassay for human chorionic gonadotropin (hCG) was developed with biotin-mediated filtration capture and silicon sensor detection. A high density of biotin on the membrane assured efficient capture of complexes containing streptavidin and analyte. Capture efficiency was not affected over a wide range of filtration flow rates or biotin concentrations. The assay utilized the pH sensing ability of the light addressable potentiometric sensor (LAPS) for the detection of urease-antibody conjugates. A LAPS reader was constructed which allowed the enzyme conjugate to be detected in approximately 1 microliter volumes. Effects from variations in detection volume were studied. 10 pg of hCG could be detected in an assay time of 20 min with four standard deviations separation from background. Comparison to a commercial RIA was made.
Olson
J D
JD
Molecular Devices Corporation, Menlo Park, CA 94025.
Panfili
P R
PR
Armenta
R
R
Femmel
M B
MB
Merrick
H
H
Gumperz
J
J
Goltz
M
M
Zuk
R F
RF
D000911:Antibodies, Monoclonal; D015374:Biosensing Techniques; D001710:Biotin; D006063:Chorionic Gonadotropin / Q000032:analysis*; D005374:Filtration; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D011863:Radioimmunoassay; D015203:Reproducibility of Results; D012828:Silicones; D014510:Urease
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D012828:Silicones; D001710:Biotin; D014510:Urease