Dry-reagent double-monoclonal assay for cystatin C

Clinical Chemistry
Noora RistiniemiKim Pettersson

Abstract

Cystatin C is a low molecular weight cysteine proteinase inhibitor whose plasma or serum concentrations have been shown to be better correlated with glomerular filtration rate than serum creatinine concentrations. Routine assays for cystatin C are based on use of polyclonal antibodies and immunoturbidimetric and nephelometric designs. This study aimed to develop a double-monoclonal immunoassay for cystatin C. We tested functionality of 42 2-site antibody combinations involving 7 monoclonal antibodies with recombinant and plasma cystatin C. We developed a heterogeneous assay using 2 antibodies selected to give the best analytical performance. The assay used a dilution step and was based on a dry-reagent, all-in-one immunoassay concept with time-resolved fluorometry. The assay was performed on an automated immunoanalyzer in single wells that contained all the required assay components. We used heparin-derived plasma samples for methodological evaluation of the assay. From a relative epitope map involving 7 cystatin C-specific antibodies, we selected a pair of antibodies for a 2-site sandwich-type dry-reagent assay. Total assay time was 15 min, and 10 microL of a 100-fold diluted sample was used. The analytical detection limit (ba...Continue Reading

References

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Citations

Jun 17, 2011·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·Noora RistiniemiAnders Christensson
Feb 22, 2012·Journal of Immunological Methods·Noora RistiniemiKim Pettersson
May 11, 2017·The Journal of Biological Chemistry·Tyler J PerlenfeinRegina M Murphy
Feb 2, 2018·Critical Reviews in Clinical Laboratory Sciences·Emil den BakkerArend Bökenkamp

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