혈중 대식세포 유주 저지 인자 측정을 위한 방사면역측정법 (Radioimmunoassay for Determination of Serum Macrophage Migration Inhibitory Factor) |
Author |
이태섭1, 신석환3, 송지인3, 우광선1, 정위섭1, 최창운2, 임상무2, |
Tae Sup Lee, Ph.D.1, Seok Hwan Shin, M.D. & Ph.D.3, Jee In Song, B.S.3, Kwang Sun Woo, B.S.1,Wee Sup Chung, B.S.1, Chang Woon Choi, M.D. & Ph.D.2, Sang Moo Lim, M.D.& Ph.D.2 |
Affiliation |
원자력의학원 싸이클로트론응용연구실1, 핵의학과2, 인하대학교 외과학교실3 Laboratory of Cyclotron Application1, Department of Nuclear Medicine2, Korea Institute of Radiological and MedicalScience (KIRAMS), Seoul; Department of Surgery3, College of Medicine, Inha University, Incheon, Korea |
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
125I-streptavidin (SA) as radiotracer to determination of rhMIF concentration. Streptavidin was labeled with 125I by
Chloramine-T method and 125I-SA was purified by ultracentrifugation. 125I-SA stability was evaluated by ITLC analysis
at 4℃ 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 125I-SA was 87%
and purified 125I-SA retained above 99% radiochemical purity. 125I-SA showed above 93% stability in 4℃ 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 125I-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 125I-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.(Korean J Nucl Med 38(6):532-539, 2004) |
Keyword |
Macrophage migration inhibitory factor (MIF), Radioimmunoassay, Streptavidin, Iodine-125, Inflammatory diseases. |
Full text Article |
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