Measurement of serum monoclonal components: comparison between densitometry and capillary zone electrophoresis
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Michele Mussap
Abstract
Quantitative measurement of serum monoclonal protein (M-protein) is one of the most important tools for monitoring disease activity in monoclonal gammopathies. The aims of this study were to evaluate serum M-protein quantification by capillary zone electrophoresis (CZE) and to compare results with those obtained by densitometric scanning of high-resolution agarose gel electrophoresis (HRE-AGE). The evaluation was carried out on 82 samples from patients with various monoclonal gammopathies. All the suspected M-proteins were confirmed and characterised by immunofixation on agarose gel (IFE). CZE was performed on a Paragon CZE™ 2000 system (Beckman Coulter). Passing-Bablok regression was: y (CZE)=1.27×(HRE-AGE)–5.21g/L. The correlation coefficient was 0.92. Bland-Altman analysis demonstrated a mean difference of −1.83g/L (95% CI −0.76 to −2.90) with clear evidence of a concentration-related bias. Densitometry gave higher values at low M-spikes (<20g/L), whereas CZE gave higher values at large M-spikes (>20g/L). The concentration-related bias was found to be independent of the immunoglobulin isotype. In conclusion, to compare previous results obtained by M-protein densitometric scanning with those obtained by direct measurement of CZE peaks, the calculation of a univocal transforming factor appears to be unreliable.
References
1. The International Myeloma Working Group. Criteria for the classification of monoclonal gammopathies, multiple myeloma and related disorders: a report of the International Myeloma Working Group. Br J Haematol 2003;121:749–57.10.1046/j.1365-2141.2003.04355.xSearch in Google Scholar
2. Kyle RA, Vincent Rajkumar S. Monoclonal gammopathies of undetermined significance. Best Pract Res Clin Haematol 2005; 18:689–707.10.1016/j.beha.2005.01.025Search in Google Scholar
3. Kyle RA, Therneau TM, Rajkumar SV, Offord JR, Larson DR, Plevak MF, et al. A long-term study of prognosis in monoclonal gammopathy of undetermined significance. N Engl J Med 2002; 346:564–9.10.1056/NEJMoa01133202Search in Google Scholar
4. Stemerman D, Papadea C, Martino-Saltzman D, O'Connell AC, Demaline B, Austin GE. Precision and reliability of paraprotein determinations by high-resolution agarose gel electrophoresis. Am J Clin Pathol 1989; 91:435–40.10.1093/ajcp/91.4.435Search in Google Scholar
5. Henskens Y, De Winter J, Pekelharing M, Ponjee G. Detection and identification of monoclonal gammopathies by capillary electrophoresis. Clin Chem 1998; 44:1184–90.10.1093/clinchem/44.6.1184Search in Google Scholar
6. Bienvenu J, Graziani MS, Arpin F, Bernon H, Blessum C, Marchetti C, et al. Multicenter evaluation of the Paragon CZE™ 2000 capillary zone electrophoresis system for serum protein electrophoresis and monoclonal component typing. Clin Chem 1998; 44:599–605.10.1093/clinchem/44.3.599Search in Google Scholar
7. Stemerman D, Papadea C, Martino-Saltzman D, O'Connell AC, Demaline B, Austin GE. Precision and reliability of paraprotein determinations by high-resolution agarose gel electrophoresis. Am J Clin Pathol 1989; 91:435–40.10.1093/ajcp/91.4.435Search in Google Scholar
8. Bablok W, Passing H, Bender R, Schneider B. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry. Part III. J Clin Chem Clin Biochem 1988; 26:783–90.Search in Google Scholar
9. Bland JM, Altman DG. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1986; i:307–10.10.1016/S0140-6736(86)90837-8Search in Google Scholar
10. Dennis PM, Biegler B, Papas R. Improved measurement of monoclonal paraproteins in serum using agarose gel electrophoresis. Ann Clin Biochem 1987; 24:73–6.10.1177/000456328702400111Search in Google Scholar PubMed
11. Kahn SN, Strony LP. Imprecision of quantification of serum protein fractions by electrophoresis on cellulose acetate. Clin Chem 1986; 32:356–7.10.1093/clinchem/32.2.356Search in Google Scholar
12. Bossuyt X. Interferences in clinical capillary zone electrophoresis of serum proteins. Electrophoresis 2004; 25:1485–7.10.1002/elps.200305820Search in Google Scholar
13. Arranz-Pena ML, Gonzalez-Sagrado M, Olmos-Linares AM, Fernandez-Garcia N, Martin-Gil FJ. Interference of iodinated contrast media in serum capillary zone electrophoresis. Clin Chem 2000; 46:736–7.10.1093/clinchem/46.5.736Search in Google Scholar
14. Bossuyt X, Mewis A, Blanckaert N. Interference of radioopaque agents in clinical capillary zone electrophoresis. Clin Chem 1999; 45:129–31.10.1093/clinchem/45.1.129Search in Google Scholar
15. Jenkins MA, Kulinskaya E, Martin HD, Guerin MD. Evaluation of serum protein separation by capillary electrophoresis: prospective analysis of 1000 specimens. J Chromatogr B Biomed Appl 1995; 672:241–51.10.1016/0378-4347(95)00224-7Search in Google Scholar
16. Katzmann JA, Clark R, Sanders E, Landers JP, Kyle RA. Prospective study of serum protein capillary zone electrophoresis and immunotyping of monoclonal proteins by immunosubtraction. Am J Clin Pathol 1998; 110:503–9.10.1093/ajcp/110.4.503Search in Google Scholar PubMed
17. Gay-Bellile C, Bengoufa D, Houze P, Le Carrer D, Benlakehal M, Bousquet B, et al. Automated multicapillary electrophoresis for analysis of human serum proteins. Clin Chem 2003; 49:1909–15.10.1373/clinchem.2003.017756Search in Google Scholar PubMed
©2006 by Walter de Gruyter Berlin New York
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