An ultrasensitive, nonisotopic immunoassay for hyaluronan using the streptavidin-biotin system

Clinica Chimica Acta; International Journal of Clinical Chemistry
A Rössler

Abstract

A time-resolved fluoroimmunoassay for measuring hyaluronan concentrations in plasma and several biological fluids is described. The solid-phase immunoassay is based on the competition between aggregation of hyaluronan with the cartilage proteoglycan monomer, followed by binding of a monoclonal antibody to keratan sulfate of the proteoglycan and a biotinylated anti-mouse IgG. Fluorescence can be measured by a time-resolved fluorometer after binding of Eu(3+)-labelled streptavidin to the biotinylated IgG. The assay is precise and correlates well (r = 0.986) with the only established radioimmunoassay known. The results show that it is essential to perform a blank run without addition of proteoglycan, as endogenous proteoglycan disturbs the measurement and causes underestimation of plasma hyaluronan. The distinguishing feature of this assay is its extreme sensitivity (< 0.24 microgram/l of plasma). The mean analytical recovery after serial dilutions and addition was 100.3 and 101.3%, the within-assay and between-assay coefficients of variation were 3.67% and 7.02%, respectively.

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Citations

May 6, 2008·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·Simon J DaviesJose C Divino Filho
Nov 22, 2011·Bio Systems·Tatsuji Takahashi, Yukio-Pegio Gunji
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Feb 27, 2010·Bio Systems·Takayuki Niizato, Yukio-Pegio Gunji
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Aug 16, 2015·Progress in Biophysics and Molecular Biology·Plamen L Simeonov
Aug 1, 2015·Studies in History and Philosophy of Science·Boris Kožnjak

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