Abstract
Background: Over the last few decades, new synthetic insulin analogues have been developed. Their measurement is of prime importance in the investigation of hypoglycaemia, but their quantification is hampered by variable cross-reactivity with many insulin assays. For clinical analysis, it has now become essential to know the potential cross-reactivity of analogues of interest.
Methods: In this work, we performed an extensive study of insulin analogue cross-reactivity using numerous human insulin immunoassays. We investigated the cross-reactivity of five analogues (lispro, aspart, glulisine, glargine, detemir) and two glargine metabolites (M1 and M2) with 16 commercial human insulin immunoassays as a function of concentration.
Results: The cross-reactivity values for insulin analogues or glargine metabolites ranged from 0% to 264%. Four assays were more specific to human insulin, resulting in negligible cross-reactivity with the analogues. However, none of the 16 assays was completely free of cross-reactivity with analogues or metabolites. The results show that analogue cross-reactivity, which varies to a large degree, is far from negligible, and should not be overlooked in clinical investigations.
Conclusions: This study has established the cross-reactivity of five insulin analogues and two glargine metabolites using 16 immunoassays to facilitate the choice of the immunoassay(s) and to provide sensitive and specific analyses in clinical routine or investigation.
The authors thank G. Coumaros and all the hormonology laboratory technicians for their technical assistance on this project as well as Klump and Guth laboratories (Strasbourg, France). We also specifically acknowledge Nathalie Heider for carefully reviewing the manuscript including English revision.
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
References
1. Sapin R. The interference of insulin antibodies in insulin immunometric assays. Clin Chem Lab Med 2002;40: 705–8.10.1515/CCLM.2002.121Search in Google Scholar PubMed
2. Sapin R. Insulin immunoassays: fast approaching 50 years of existence and still calling for standardization. Clin Chem 2007;53:810–2.10.1373/clinchem.2006.084012Search in Google Scholar PubMed
3. Manley SE, Stratton IM, Clark PM, Luzio SD. Comparison of 11 human insulin assays: implications for clinical investigation and research. Clin Chem 2007;53:922–32.10.1373/clinchem.2006.077784Search in Google Scholar PubMed
4. Marcovina S, Bowsher RR, Miller WG, Staten M, Myers G, Caudill SP, et al. Standardization of insulin immunoassays: report of the American Diabetes Association Workgroup. Clin Chem 2007;53:711–6.10.1373/clinchem.2006.082214Search in Google Scholar PubMed
5. Rodriguez-Cabaleiro D, Van Uytfanghe K, Stove V, Fiers T, Thienpont LM. Pilot study for the standardization of insulin immunoassays with isotope dilution liquid chromatography/tandem mass spectrometry. Clin Chem 2007;53:1462–9.10.1373/clinchem.2007.088393Search in Google Scholar PubMed
6. Hirsch IB. Insulin analogues. N Engl J Med 2005;352:174–83.10.1056/NEJMra040832Search in Google Scholar PubMed
7. Thevis M, Thomas A, Delahaut P, Bosseloir A, Schanzer W. Qualitative determination of synthetic analogues of insulin in human plasma by immunoaffinity purification and liquid chromatography-tandem mass spectrometry for doping control purposes. Anal Chem 2005;77:3579–85.10.1021/ac050066iSearch in Google Scholar PubMed
8. Thevis M, Thomas A, Delahaut P, Bosseloir A, Schanzer W. Doping control analysis of intact rapid-acting insulin analogues in human urine by liquid chromatography-tandem mass spectrometry. Anal Chem 2006;78:1897–903.10.1021/ac052095zSearch in Google Scholar PubMed
9. Thevis M, Thomas A, Schanzer W. Insulin. Handb Exp Pharmacol 2010;195:209–26.10.1007/978-3-540-79088-4_10Search in Google Scholar PubMed
10. Thomas A, Schanzer W, Delahaut P, Thevis M. Sensitive and fast identification of urinary human, synthetic and animal insulin by means of nano-UPLC coupled with high-resolution/high-accuracy mass spectrometry. Drug Test Anal 2009;1:219–27.10.1002/dta.35Search in Google Scholar PubMed
11. Thomas A, Thevis M, Delahaut P, Bosseloir A, Schanzer W. Mass spectrometric identification of degradation products of insulin and its long-acting analogues in human urine for doping control purposes. Anal Chem 2007;79:2518–24.10.1021/ac062037tSearch in Google Scholar
12. Moriyama M, Hayashi N, Ohyabu C, Mukai M, Kawano S, Kumagai S. Performance evaluation and cross-reactivity from insulin analogs with the ARCHITECT insulin assay. Clin Chem 2006;52:1423–6.10.1373/clinchem.2005.065995Search in Google Scholar
13. Owen WE, Roberts WL. Cross-reactivity of three recombinant insulin analogs with five commercial insulin immunoassays. Clin Chem 2004;50:257–9.10.1373/clinchem.2003.026625Search in Google Scholar
14. Vieira JG, Tachibana TT, Ferrer CM, Reis AF. Cross-reactivity of new insulin analogs in insulin assays. Arq Bras Endocrinol Metabol 2007;51:504–5.10.1590/S0004-27302007000300022Search in Google Scholar
15. Krull I, Sahli R, Diem P, Stettler C. Variability in cross-reactivity of novel insulin analogues in immunometric insulin assays. Diabet Med 2009;26:1075–6.10.1111/j.1464-5491.2009.02815.xSearch in Google Scholar
16. Neal JM, Han W. Insulin immunoassays in the detection of insulin analogues in factitious hypoglycemia. Endocr Pract 2008;14:1006–10.10.4158/EP.14.8.1006Search in Google Scholar
17. Agin A, Jeandidier N, Gasser F, Grucker D, Sapin R. Use of insulin immunoassays in clinical studies involving rapid-acting insulin analogues: Bi-insulin IRMA preliminary assessment. Clin Chem Lab Med 2006;44:1379–82.10.1515/CCLM.2006.257Search in Google Scholar
18. Cao Y, Smith WC, Bowsher RR. A sensitive chemiluminescent enzyme immunoassay for the bioanalysis of carboxyl-terminal B-chain analogues of human insulin. J Pharm Biomed Anal 2001;26:53–61.10.1016/S0731-7085(01)00345-4Search in Google Scholar
19. Glenn C, Armston A. Cross-reactivity of 12 recombinant insulin preparations in the Beckman Unicel DxI 800 insulin assay. Ann Clin Biochem 2010;47:264–6.10.1258/acb.2010.010002Search in Google Scholar PubMed
20. Kim S, Yun YM, Hur M, Moon HW, Kim JQ. The effects of anti-insulin antibodies and cross-reactivity with human recombinant insulin analogues in the E170 insulin immunometric assay. Korean J Lab Med 2011;31:22–9.10.3343/kjlm.2011.31.1.22Search in Google Scholar PubMed PubMed Central
21. Sapin R, Le Galudec V, Gasser F, Pinget M, Grucker D. Elecsys insulin assay: free insulin determination and the absence of cross-reactivity with insulin lispro. Clin Chem 2001;47: 602–5.10.1093/clinchem/47.3.602Search in Google Scholar
22. Song D, Davidson J. Cross-reactivity of Actrapid and three insulin analogues in the Abbott IMx insulin immunoassay. Ann Clin Biochem 2007;44:197–8.10.1258/000456307780118109Search in Google Scholar
23. Armitage P, Berry G. Statistical methods in medical research, 3rd ed. Oxford: Blackwell, 1994.Search in Google Scholar
24. Miller WG, Thienpont LM, Van Uytfanghe K, Clark PM, Lindstedt P, Nilsson G, et al. Toward standardization of insulin immunoassays. Clin Chem 2009;55:1011–8.10.1373/clinchem.2008.118380Search in Google Scholar
25. Kuerzel GU, Shukla U, Scholtz HE, Pretorius SG, Wessels DH, Venter C, et al. Biotransformation of insulin glargine after subcutaneous injection in healthy subjects. Curr Med Res Opin 2003;19:34–40.10.1185/030079902125001416Search in Google Scholar
26. Sommerfeld MR, Muller G, Tschank G, Seipke G, Habermann P, Kurrle R, et al. In vitro metabolic and mitogenic signaling of insulin glargine and its metabolites. PLoS One 2010;5:e9540.10.1371/journal.pone.0009540Search in Google Scholar
27. Morovat A. Detection of Actrapid by insulin assays. Ann Clin Biochem 2007;44:315–6.10.1258/000456307780480819Search in Google Scholar
28. Marks A, Yip C, Wilson S. Characterization of two epitopes on insulin using monoclonal antibodies. Mol Immunol 1985;22:285–90.10.1016/0161-5890(85)90163-4Search in Google Scholar
29. Allauzen S, Joly S, Granier C, Molina F, Bouix O, Pau B, et al. Immunoanalysis of human insulin using monoclonal antibodies reveals antigenicity of evolutionarily conserved residues. Mol Immunol 1995;32:27–36.10.1016/0161-5890(94)00133-LSearch in Google Scholar
30. Schroer JA, Bender T, Feldmann RJ, Kim KJ. Mapping epitopes on the insulin molecule using monoclonal antibodies. Eur J Immunol 1983;13:693–700.10.1002/eji.1830130902Search in Google Scholar PubMed
31. Owens DR. Glargine and cancer: can we now suggest closure? Diabetes Care 2012;35:2426–8.10.2337/dc12-1968Search in Google Scholar PubMed PubMed Central
32. Pierre-Eugene C, Pagesy P, Nguyen TT, Neuille M, Tschank G, Tennagels N, et al. Effect of insulin analogues on insulin/IGF1 hybrid receptors: increased activation by glargine but not by its metabolites M1 and M2. PLoS One 2012;7:e41992.10.1371/journal.pone.0041992Search in Google Scholar PubMed PubMed Central
33. Bolli GB, Hahn AD, Schmidt R, Eisenblaetter T, Dahmen R, Heise T, et al. Plasma exposure to insulin glargine and its metabolites M1 and M2 after subcutaneous injection of therapeutic and supratherapeutic doses of glargine in subjects with type 1 diabetes. Diabetes Care 2012;35:2626–30.10.2337/dc12-0270Search in Google Scholar PubMed PubMed Central
34. Lucidi P, Porcellati F, Rossetti P, Candeloro P, Andreoli AM, Cioli P, et al. Metabolism of insulin glargine after repeated daily subcutaneous injections in subjects with type 2 diabetes. Diabetes Care 2012;35:2647–9.10.2337/dc12-0271Search in Google Scholar PubMed PubMed Central
35. Miller JJ, Valdes R. Methods for calculating cross-reactivity in immunoassay. J Clin Immunoassay 1992;15:97–107.Search in Google Scholar
36. Miller JJ, Valdes R, Jr. Approaches to minimizing interference by cross-reacting molecules in immunoassays. Clin Chem 1991;37:144–53.10.1093/clinchem/37.2.144Search in Google Scholar
37. Burge MR, McLeod J, Bowsher RR, Schade DS. Validity of Coat-A-Count insulin RIA kit for quantifying total and free humalog. Clin Chem 1996;42:777.10.1093/clinchem/42.5.777Search in Google Scholar
©2014 by Walter de Gruyter Berlin Boston
