Inorganic pyrophosphatase-labelled enzyme immunoassay for beta 2-microglobulin.
beta 2-Microglobulin was purified from human peritoneal dialysate by ultrafiltration, gel chromatography and DEAE-high performance chromatography. Anti-beta 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 microgram/l (3 x SD) and the assay was linear up to absorbance values of around 2.0. No hook effect occurred in any putative concentrations of beta 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 degrees C. It was decided that inorganic pyrophosphatase is a label of choice for ELISA.
Peuravuori
H
H
Department of Pathology, University of Turku, Finland. Heikki.Peuravuori@utu.fi
Kunelius
R
R
Eskola
J
J
Korpela
T
T
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011755:Pyrophosphatases / Q000378:metabolism*; D001613:beta 2-Microglobulin / Q000302:isolation & purification*
D001613:beta 2-Microglobulin; D011755:Pyrophosphatases