Abstract
β 2-Microglobulin was purified from human peritoneal dialysate by ultrafiltration, gel chromatography and DEAE-high performance chromatography. Anti-β 2-monoclonal antibodies were developed in mice and a pair of the antibodies was utilized to develop an enzyme-linked immunosorbent assay (ELISA) kit for the analyte with utilizing a new enzyme label, inorganic pyrophosphatase (EC 3.6.1.1). The sensitivity of the assay was 0.6 μg/l (3×SD) and the assay was linear up to absorbance values of around 2.0. No hook effect occurred in any putative concentrations of β 2-microglobulin in serum. The precision of the assay of one run varied within 5–7% CV and the interassay precision was 2.8–8.6% CV, while the recovery was 99.2±6.0%. Excellent correlation occurred against an established radioimmunoassay method. All the used reagents were storable for a minimum of 1 year at +4°C. It was decided that inorganic pyrophosphatase is a label of choice for ELISA.
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