Ultrasensitive time-resolved immunofluorometry of human albumin in urine using monoclonal antibodies--a new assay for microalbuminuria.
We describe a two-site, sandwich methodology for human albumin in urine. In the assay, albumin binds to a solid-phase monoclonal antibody and to another monoclonal that is biotinylated. The immunocomplex is then quantified by adding streptavidin which is labelled with an europium chelator, using time-resolved fluorometry. The assay is extremely sensitive (less than 1 microgram/L and specific. A sample predilution of 251-fold or more is needed before analysis. The analytical parameters studied (precision, recovery, linearity, comparisons) were found to be satisfactory. The assay is simple to perform and is proposed as a non-isotopic alternative to radioimmunoassay for the quantification of small amounts of albumin in urine for the purpose of assessing microalbuminuria.
Department of Clinical Biochemistry, Toronto Western Hospital, Ontario, Canada.
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Diamandis
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Ogilvie
D000419:Albuminuria / Q000175:diagnosis*; D000911:Antibodies, Monoclonal; D015200:Fluoroimmunoassay; D006801:Humans; D011933:Reagent Kits, Diagnostic; D012016:Reference Values; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D011933:Reagent Kits, Diagnostic