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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

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CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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Volume 57, Issue 11


Variable and inaccurate serum IgG4 levels resulting from lack of standardization in IgG subclass assay calibration

Luca Bernasconi / Esther Mundwiler / Stephan Regenass / Vincent Aubert / Angelika Hammerer-Lercher / Ingmar Heijnen
Published Online: 2019-06-11 | DOI: https://doi.org/10.1515/cclm-2019-0261



The quantification of serum IgG4 is commonly performed during the diagnostic workup of IgG4-related diseases (IgG4-RD). According to recent literature, IgG4 values above 1.35 g/L are characteristic of IgG4-RD and support its diagnosis at initial presentation. The purpose of this study was to evaluate comparability and accuracy of the two main commercially available IgG4 assays (Siemens Healthineers and The Binding Site).


Method comparison was performed for IgG and IgG subclasses using a collective of selected samples with elevated serum IgG4. In addition, we assessed the accuracy of both assays using purified polyclonal and monoclonal IgG4 preparations.


Our data show significant discrepancies between the two IgG subclass assays for the measurement of IgG4 and, to a lesser extent, IgG3.


The lack of standardization between the two main providers of commercially available IgG4 assays leads to significant inter-assay result discrepancies, which might potentially cause unnecessary clinical workup. We conclude that serum IgG4 assay-specific decision limits, and not an assay-independent single cut-off level for IgG4 (e.g. 1.35 g/L), should be used when assessing patients for IgG4-RD. An internationally recognized, certified reference material for IgG subclasses is urgently needed, and assay manufactures are encouraged to undertake steps toward standardization of measurements of IgG4 and other IgG subclasses.

This article offers supplementary material which is provided at the end of the article.

Keywords: IgG subclasses; IgG4-related disease; IgG4 quantification; standardization


  • 1.

    Stone JH, Zen Y, Deshpande V. IgG4-related disease. N Engl J Med 2012;366:539–51.CrossrefPubMedGoogle Scholar

  • 2.

    Ghazale A, Chari ST, Smyrk TC, Levy MJ, Topazian MD, TakahashiN, et al. Value of serum IgG4 in the diagnosis of autoimmune pancreatitis and in distinguishing it from pancreatic cancer. Am J Gastroenterol 2007;102:1646–53.CrossrefWeb of ScienceGoogle Scholar

  • 3.

    Hamano H, Kawa S, Horiuchi A, Unno H, Furuya N, Akamatsu T, et al. High serum IgG4 concentrations in patients with sclerosing pancreatitis. N Engl J Med 2001;344:732–8.CrossrefPubMedGoogle Scholar

  • 4.

    Masaki Y, Kurose N, Yamamoto M, Takahashi H, Saeki T, Azumi A, et al. Cutoff values of serum IgG4 and histopathological IgG4+ plasma cells for diagnosis of patients with IgG4-related disease. Int J Rheumatol 2012;2012:580814.PubMedGoogle Scholar

  • 5.

    Vlug A, Nieuwenhuys EJ, van Eijk RV, Geertzen HG, van Houte AJ. Nephelometric measurements of human IgG subclasses and their reference ranges. Ann Biol Clin (Paris) 1994;52:561–7.PubMedGoogle Scholar

  • 6.

    Xu WL, Ling YC, Wang ZK, Deng F. Diagnostic performance of serum IgG4 level for IgG4-related disease: a meta-analysis. Sci Rep 2016;6:32035.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 7.

    Klein F, Skvaril F, Vermeeren R, Vlug A, Duimel WJ. The quantification of human IgG subclasses in reference preparations. Clin Chim Acta 1985;150:119–27.PubMedCrossrefGoogle Scholar

  • 8.

    Schauer U, Stemberg F, Rieger CH, Borte M, Schubert S, Riedel F, et al. IgG subclass concentrations in certified reference material 470 and reference values for children and adults determined with the binding site reagents. Clin Chem 2003;49:1924–9.CrossrefPubMedGoogle Scholar

  • 9.

    Bossuyt X, Marien G, Meyts I, Proesmans M, De Boeck K. Determination of IgG subclasses: a need for standardization. J Allergy Clin Immunol 2005;115:872–4.CrossrefPubMedGoogle Scholar

  • 10.

    Parker AR, Hughes RG, Mead GP, Carr-Smith HD. Reply to “pediatric reference intervals for immunoglobulin G and its subclasses with Siemens immunonephelometric assays”. Clin Biochem 2011;44:745–6.Web of ScienceCrossrefGoogle Scholar

  • 11.

    Wilson C, Ebling R, Henig C, Adler T, Nicolaevski R, Barak M, et al. Significant, quantifiable differences exist between IgG subclass standards WHO67/97 and ERM-DA470k and can result in different interpretation of results. Clin Biochem 2013;46:1751–5.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 12.

    Ludwig-Kraus B, Kraus FB. Similar but not consistent: revisiting the pitfalls of measuring IgG subclasses with different assays. J Clin Lab Anal 2017;31:e22146.CrossrefWeb of ScienceGoogle Scholar

  • 13.

    Sarnago A, Pascual RM, Moreno MJ, Laiz B, Fuster O. IgG subclasses quantitation: analytical performance of The Binding Site SPAPLUS(R) human assay and comparison with Siemens BNII(R) assay. Clin Biochem 2018;51:85–9.CrossrefPubMedGoogle Scholar

  • 14.

    Ladwig PM, Barnidge DR, Snyder MR, Katzmann JA, Murray DL. Quantification of serum IgG subclasses by use of subclass-specific tryptic peptides and liquid chromatography-tandem mass spectrometry. Clin Chem 2014;60:1080–8.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 15.

    van der Gugten G, DeMarco ML, Chen LY, Chin A, Caruthers M, Holmes DT, et al. Resolution of spurious immunonephelometric IgG subclass measurement discrepancies by LC-MS/MS. Clin Chem 2018;64:735–42.Web of ScienceCrossrefPubMedGoogle Scholar

  • 16.

    Umehara H, Okazaki K, Nakamura T, Satoh-Nakamura T, Nakajima A, Kawano M, et al. Current approach to the diagnosis of IgG4-related disease – combination of comprehensive diagnostic and organ-specific criteria. Mod Rheumatol 2017;27:381–91.Web of ScienceCrossrefGoogle Scholar

  • 17.

    Wallace ZS, Zhang Y, Perugino CA, Naden R, Choi HK, Stone JH, et al. Clinical phenotypes of IgG4-related disease: an analysis of two international cross-sectional cohorts. Ann Rheum Dis 2019;78:406–12.Web of SciencePubMedCrossrefGoogle Scholar

  • 18.

    Carruthers MN, Khosroshahi A, Augustin T, Deshpande V, Stone JH. The diagnostic utility of serum IgG4 concentrations in IgG4-related disease. Ann Rheum Dis 2015;74:14–8.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 19.

    Culver EL, Sadler R, Simpson D, Cargill T, Makuch M, Bateman AC, et al. Elevated serum IgG4 levels in diagnosis, treatment response, organ involvement, and relapse in a prospective IgG4-related disease UK cohort. Am J Gastroenterol 2016;111:733–43.CrossrefWeb of ScienceGoogle Scholar

About the article

Corresponding author: Luca Bernasconi, PhD, Institute of Laboratory Medicine, Kantonsspital Aarau AG, Tellstr. 25, 5001 Aarau, Switzerland, Phone: +41 62 838 53 15, Fax: +41 62 838 53 99

Received: 2019-03-07

Accepted: 2019-05-15

Published Online: 2019-06-11

Published in Print: 2019-10-25

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 11, Pages 1777–1783, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2019-0261.

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