Flow injection renewable surface immunoassay: a new approach to immunoanalysis with fluorescence detection.
This paper introduces a new methodology of carrying out heterogeneous immunoassays automatically, using a flow injection technique on a renewable surface. Flow injection renewable surface immunoassay (FIRSI) relies on the use of a minute amount of beads to form a reactive surface, which is interrogated by fluorescence spectrometry. Following the assay, on-line regeneration normally used in flow based immunoassays is avoided by fluidically removing the spent reactive surface and replacing it with a new layer of beads. This allows the monitoring of antibody-antigen binding at its early stages, dramatically increases the sampling frequency of a serial assay, and eliminates the problems associated with a decrease in surface reactivity caused by repetitive use. A model system utilizing anti-mouse IgG1-coated beads and mouse IgG1 protein is used to characterize the method with respect to reproducibility, flow rate, contact time, and amount of beads.
Department of Chemistry BG-10, University of Washington, Seattle 98195.
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Pollema
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Ruzicka
D000818:Animals; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D013050:Spectrometry, Fluorescence
D007074:Immunoglobulin G