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Radioimmunoassay for Determination of Serum Macrophage Migration Inhibitory Factor  

Lee, Tae-Sup (Laboratory of Cyclotron Application, Korea Institute of Radiological and Medical Science (KIRAMS))
Shin, Seok-Hwan (Department of Surgery, College of Medicine, Inha University)
Song, Jee-In (Department of Surgery, College of Medicine, Inha University)
Woo, Kwang-Sun (Laboratory of Cyclotron Application, Korea Institute of Radiological and Medical Science (KIRAMS))
Chung, Wee-Sup (Department of Nuclear Medicine, Korea Institute of Radiological and Medical Science (KIRAMS))
Choi, Chang-Woon (Department of Nuclear Medicine, Korea Institute of Radiological and Medical Science (KIRAMS))
Lim, Sang-Moo (Laboratory of Cyclotron Application, Korea Institute of Radiological and Medical Science (KIRAMS))
Publication Information
The Korean Journal of Nuclear Medicine / v.38, no.6, 2004 , pp. 532-539 More about this Journal
Abstract
Purpose: There has been a renewal of interest in Macrophage migration inhibitory factor (MIF), especially correlation in pathogenesis of sepsis by many infectious diseases and in regulation of host inflammatory and immune response. We developed immunoradiometric assay (IRMA) to determine serum human MIF concentration. Materials and Methods: The IRMA system utilizes solid phase bound monoclonal anti-recombinant human MIF (rhMIF) antibody as a capture antibody, biotinylated polyclonal anti-rhMIF antibody as a detector antibody. We applied with rhMIF that concentration of standard solutions increased from 0 ng/ml to 100 ng/ml. We used $^{125}I$-streptavidin (SA) as radiotracer to determination of rhMIF concentration. Streptavidin was labeled with $^{125}I$ by Chloramine-T method and $^{125}I$-SA was purified by ultracentrifugation. $^{125}I$-SA stability was evaluated by ITLC analysis at $4^{\circ}C$ and room temperatures until 60days. To validate IRMA system for MIF, we experimented intra-assay and inter-assay coefficients of variation, recovery test and dilution test. Results: Radiolabeling yield of $^{125}I$-SA was 87% and purified $^{125}I$-SA retained above 99% radiochemical purity. $^{125}I$-SA showed above 93% stability in $4^{\circ}C$ until 60days that it is good for immunoradiometric assay as radiotracer. Plotted standard dose response curve showed that increased concentration of rhMIF linearly correlated (R2=0.99) with bound radioactivity of $^{125}I$-SA. The highest intra- and inter-assay coefficients of variation were 5.5% and 7.6%, respectively. The average of recovery of MIF in samples was 102%. In dilution test, linear response curves were obtained (R2=0.97). Conclusion: Radioimmunoassay using $^{125}I$-SA as radiotracer thought to be useful for the determination of serum MIF concentration, and further, its data will be used to evaluate the correlation between clinical significance and serum MIF concentration in patients with various inflammatory diseases.
Keywords
Macrophage migration inhibitory factor (MIF); Radioimmunoassay; Streptavidin; $Iodine-^{125}$; Inflammatory diseases;
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