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Validation of sclerostin ELISAs in HD patients

DISCOVERIES (ISSN 2359-7232), 2016, January-March issue


Mause SF, Deck A, Hennies M, Kaesler N, Evenepoel P, Janssen U, Brandenburg VM. Validation of commercially available ELISAs for the detection of circulating sclerostin in hemodialysis patients. Discoveries 2016, January-March; 4(1): e55. DOI: 10.15190/d.2016.2

Submitted: March 25th, 2016Revised: March 30th, 2016Accepted: Martch 31st, 2016Published: March 31st, 2016;

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Validation of commercially available ELISAs for the detection of circulating sclerostin in hemodialysis patients

Sebastian F. Mause (*,1), Annika Deck (1), Mark Hennies (2), Nadine Kaesler (3), Pieter Evenepoel (4), William A. Boisvert (5), Ulf Janssen (#,6), Vincent M. Brandenburg (#,1)

(1) Department of Cardiology, University Hospital of the RWTH Aachen, Germany

(2) TECOmedical AG, Sissach, Switzerland

(3) Department of Nephrology, University Hospital of the RWTH Aachen, Aachen, Germany

(4) Department of Medicine, University Hospital Leuven, Leuven, Belgium

(5) Center for Cardiovascular Research, John A. Burns School of Medicine, University of Hawaii, Honolulu, USA

(6) Department of Nephrology, Klinikum Maria-Hilf, Mönchengladbach, Germany

# These authors contributed equally to this work

*Correspondence to: Sebastian F. Mause, MD, Department of Cardiology, Pauwelsstraße 30, Dā€52074 Aachen, University Hospital RWTH Aachen, Germany; Phone: +49 (0)241/80 35223; Fax: +49 (0)241/80 82446; Email: smause@ukaachen.de


BACKGROUND: Sclerostin is an endocrine regulator in chronic kidney disease – mineral and bone disorder (CKD-MBD). Validation of assay comparability and pre-analytical handling is mandatory for establishment of sclerostin as a biomarker. 

METHODS: Blood samples (serum, EDTA, heparin and citrate plasma) were obtained from 12 hemodialysis (HD) patients after the long dialysis interval. Passing-Bablok regression analysis and Bland-Altman difference plots were used to evaluate the agreement between sclerostin levels measured with two commercially available ELISAs from TECOmedical and Biomedica.

RESULTS: Independent of the sample type, the agreement of the two assays was poor with a strong proportional but no systematic bias. Compared to the TECOmedical assay, the Biomedica test yielded almost 2-fold higher sclerostin values throughout all sample types. Spike recovery and linear dilution studies revealed a higher accuracy of the TECOmedical assay (97% and 96%) compared tothe Biomedica assay (118% and 78%). Sclerostin levels were stable within 4 hours after sample collection, in particular when analyzed in plasma. In contrast to the Biomedica assay, the TECOmedical showed a systematic but no proportional bias between serum and plasma samples with higher values for plasma samples. Among the 3 different plasma samples no systematic error could be documented.

CONCLUSION: Careful consideration of the pre-analytical handling and comparative assay validation are necessary to facilitate a more differentiated interpretation of studies reporting circulating sclerostin levels. The presence of a proportional bias demonstrates that in HD patients the two ELISAs for measuring sclerostin should not be used interchangeably. Furthermore, caution is necessary when comparing sclerostin results obtained from different blood sample types.


Access full text of the manuscript here:    Supplementary Information (pdf)


1. Moe S, Drueke T, Cunningham J, et al. Definition, evaluation, and classification of renal osteodystrophy: a position statement from Kidney Disease: Improving Global Outcomes (KDIGO). Kidney Int. 2006;69:1945-1953.

2. Winkler DG, Sutherland MK, Geoghegan JC, et al. Osteocyte control of bone formation via sclerostin, a novel BMP antagonist. EMBO J. 2003;22:6267-6276.

3. Brandenburg VM., D'Haese P, Deck A, et al. From skeletal to cardiovascular disease in 12 steps-the evolution of sclerostin as a major player in CKD-MBD. Pediatr Nephrol. 2016;31:195-206

4. Sabbagh Y, Graciolli FG, O'Brien S, et al. Repression of osteocyte Wnt/beta-catenin signaling is an early event in the progression of renal osteodystrophy. J Bone Miner Res. 2012;27:1757-1772.

5. Ishimura E, Okuno S, Ichii M, et al. Relationship between serum sclerostin, bone metabolism markers, and bone mineral density in maintenance hemodialysis patients. J Clin Endocrinol Metab. 2014;99:4315-4320.

6. Drechsler C, Evenepoel P, Vervloet MG, et al. High levels of circulating sclerostin are associated with better cardiovascular survival in incident dialysis patients: results from the NECOSAD study. Nephrol Dial Transplant. 2015;30:288-293.

7. Goncalves FL, Elias RM, dos Reis LM, et al. Serum sclerostin is an independent predictor of mortality in hemodialysis patients. BMC Nephrol. 2014; 15:190.

8. Claes KJ, Viaene L, Heye S, et al. Sclerostin: Another vascular calcification inhibitor? J Clin Endocrinol Metab. 2013;98:3221-3228.

9. Brandenburg VM, Kramann R, Koos R, et al. Relationship between sclerostin and cardiovascular calcification in hemodialysis patients: a cross-sectional study. BMC Nephrol. 2013;14:219.

10. Pelletier S, Confavreux CB, Haesebaert J, et al. Serum sclerostin: the missing link in the bone-vessel cross-talk in hemodialysis patients? Osteoporos Int. 2015;26:2165-2174

11. Chau CH, Rixe O, McLeod H, Figg WD. Validation of analytic methods for biomarkers used in drug development. Clin Cancer Res. 2008;14:5967-5976.

12. Cantor T, Yang Z, Caraiani N, Ilamathi E. Lack of comparability of intact parathyroid hormone measurements among commercial assays for end-stage renal disease patients: implication for treatment decisions. Clin Chem. 2006;52:1771-1776.

13. McNulty M, Singh RJ, Li X, Bergstralh EJ, Kumar R. Determination of serum and plasma sclerostin concentrations by enzyme-linked immunoassays. J Clin Endocrinol Metab 2011;96:E1159-1162.

14. Durosier C, van Lierop A, Ferrari S, et al. Association of circulating sclerostin with bone mineral mass, microstructure, and turnover biochemical markers in healthy elderly men and women. J Clin Endocrinol Metab. 2013;98:3873-3883.

15. Costa AG, Cremers S, Dworakowski E, Lazaretti-Castro M, Bilezikian JP. Comparison of two commercially available ELISAs for circulating sclerostin. Osteoporos Int. 2014;25:1547-1554.

16. Clarke BL, Drake MT.  Clinical utility of serum sclerostin measurements. Bonekey Rep. 2013;2:361.

17. Bland JM, Altman DG. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet. 1986;1:307-310.

18. Moysés RM, Jamal SA, Graciolli FG, et al. Can we compare serum sclerostin results obtained with different assays in hemodialysis patients?: Int Urol Nephrol. 2015;47:847-50.

19. Boterman M, Doig M, Breda M, et al. Recommendations on the interpretation of the new European Medicines Agency Guideline on Bioanalytical Method Validation by Global CRO Council for Bioanalysis (GCC). Bioanalysis. 2012;4:651-660.

20. Zaki R, Bulgiba A, Ismail R, Ismail NA. Statistical methods used to test for agreement of medical instruments measuring continuous variables in method comparison studies: a systematic review. PLoS One. 2012;7:e37908.

21. Passing H, Bablok. A new biometrical procedure for testing the equality of measurements from two different analytical methods. Application of linear regression procedures for method comparison studies in clinical chemistry, Part I. J Clin Chem Clin Biochem. 1983;21:709-720.

22.Veverka V, Henry AJ, Slocombe PM, et al. Characterization of the structural features and interactions of sclerostin: molecular insight into a key regulator of Wnt-mediated bone formation. J Biol Chem. 2009;284:10890-10900.

23. Hernandez P, Whitty C, John Wardale R, Henson FM. New insights into the location and form of sclerostin. Biochem Biophys Res Commun. 2014;446:1108-1113.

24. Miles RR, Roberts RF, Putnam AR, Roberts WL. Comparison of serum and heparinized plasma samples for measurement of chemistry analytes. Clin Chem. 2004;50:1704-1706.

25. van Lierop AH, van der Eerden AW, et al. Circulating sclerostin levels are decreased in patients with endogenous hypercortisolism and increase after treatment. J Clin Endocrinol Metab. 2012;97:E1953-1957.

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