Enzyme-linked immunosorbent assays in a chromatographic format.
This paper describes the execution of enzyme-linked immunosorbent assays (ELISA), (i) in immunosorbent columns with antibodies immobilized on porous particles, (ii) where samples and reagents are metered by valves and syringe pumps, (iii) samples, reagents, substrate, and wash buffers are transported into or through the system by high pressure liquid chromatography pumps, and (iv) enzyme reaction product is detected by absorbance. The normal protocol used for ELISA was modified in that antigen was complexed with enzyme conjugated antibody in the autosampler and an aliquot introduced into the system where it was transported to the immunosorbent and captured. Substrate was subsequently pumped into the immunosorbent bed and the product swept to an absorbance detector for quantitation.
Johns
M A
MA
PerSeptive Biosystems, Inc., Framingham, MA 01701, USA.
Rosengarten
L K
LK
Jackson
M
M
Regnier
F E
FE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001331:Automation / Q000295:instrumentation*; D006063:Chorionic Gonadotropin / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D051379:Mice; D016017:Odds Ratio; D010655:Phenylenediamines / Q000378:metabolism; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013050:Spectrometry, Fluorescence; D013972:Thyrotropin / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D010655:Phenylenediamines; D011933:Reagent Kits, Diagnostic; C034193:1,2-diaminobenzene; D013972:Thyrotropin; D006735:Horseradish Peroxidase; D016650:Fluorescein-5-isothiocyanate