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Clinical Chemistry, Vol 42, 586-592, Copyright © 1996 by American Association for Clinical Chemistry
BW Hollis, JQ Kamerud, A Kurkowski, J Beaulieu and JL Napoli
Department of Pediatrics, Medical University of South Carolina, Charleston, 29425, USA.
We report here the first RIA for 1,25-dihydroxyvitamin D utilizing a radioiodinated (125I) tracer. This is also the first validated RIA for 1,25-dihydroxyvitamin D [1,25(OH)2D] that does not require sample prepurification by HPLC before the binding assay. The assay involves acetonitrile extraction, treatment of the crude extract supernate with sodium periodate, extraction and purification of endogenous 1,25(OH)2D by solid-phase chromatography, and finally, quantification by RIA. Calibrators were prepared in stripped human serum and processed exactly the same as samples, eliminating the need for internal control for procedural losses of endogenous 1,25(OH)2D. The assay consists of a 2-h room temperature incubation with the primary antibody, a 20-min incubation with a second antibody, and separation of bound from free by centrifugation. Assay results can be in hand with 5 h. The detection limit of the assay is 2.4 ng/L 1,25-dihydroxyvitamin D3. Results compare well with those from an accepted radioreceptor assay. Sample pretreatment with sodium periodate is absolutely essential before quantification by RIA; otherwise, concentrations of endogenous 1,25(OH)2D may be greatly overestimated.
The following articles in journals at HighWire Press have cited this article:
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 L. Szeles, G. Keresztes, D. Torocsik, Z. Balajthy, L. Krenacs, S. Poliska, A. Steinmeyer, U. Zuegel, M. Pruenster, A. Rot, et al. 1,25-Dihydroxyvitamin D3 Is an Autonomous Regulator of the Transcriptional Changes Leading to a Tolerogenic Dendritic Cell Phenotype J. Immunol., February 15, 2009; 182(4): 2074 - 2083. [Abstract] [Full Text] [PDF]
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 K. M. Carnagey, E. J. Huff-Lonergan, A. Trenkle, A. E. Wertz-Lutz, R. L. Horst, and D. C. Beitz Use of 25-hydroxyvitamin D3 and vitamin E to improve tenderness of beef from the longissimus dorsi of heifers J Anim Sci, July 1, 2008; 86(7): 1649 - 1657. [Abstract] [Full Text] [PDF]
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B. W. Hollis Phase Switching SPE for Faster 1,25-dihydroxyvitamin D Analysis Clin. Chem., February 1, 2008; 54(2): 446 - 447. [Full Text] [PDF]
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M. R. Waldron, B. J. Nonnecke, T. Nishida, R. L. Horst, and T. R. Overton Effect of Lipopolysaccharide Infusion on Serum Macromineral and Vitamin D Concentrations in Dairy Cows J Dairy Sci, November 1, 2003; 86(11): 3440 - 3446. [Abstract] [Full Text] [PDF]
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 M. J. Horwitz, M. B. Tedesco, S. M. Sereika, B. W. Hollis, A. Garcia-Ocana, and A. F. Stewart Direct Comparison of Sustained Infusion of Human Parathyroid Hormone-Related Protein-(1-36) [hPTHrP-(1-36)] Versus hPTH-(1-34) on Serum Calcium, Plasma 1,25-Dihydroxyvitamin D Concentrations, and Fractional Calcium Excretion in Healthy Human Volunteers J. Clin. Endocrinol. Metab., April 1, 2003; 88(4): 1603 - 1609. [Abstract] [Full Text] [PDF]
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P. R. Holt, N. Arber, B. Halmos, K. Forde, H. Kissileff, K. A. McGlynn, S. F. Moss, K. Fan, K. Yang, and M. Lipkin Colonic Epithelial Cell Proliferation Decreases with Increasing Levels of Serum 25-Hydroxy Vitamin D Cancer Epidemiol. Biomarkers Prev., January 1, 2002; 11(1): 113 - 119. [Abstract] [Full Text]
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E. A. Platz, S. E. Hankinson, B. W. Hollis, G. A. Colditz, D. J. Hunter, F. E. Speizer, and E. Giovannucci Plasma 1,25-Dihydroxy- and 25-Hydroxyvitamin D and Adenomatous Polyps of the Distal Colorectum Cancer Epidemiol. Biomarkers Prev., October 1, 2000; 9(10): 1059 - 1065. [Abstract] [Full Text]
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