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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


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

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

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1437-4331
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Volume 57, Issue 2

Issues

Different immunoreactivity of monomers and dimers makes automated free light chains assays not equivalent

Laura Caponi
  • Corresponding author
  • Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy
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  • Other articles by this author:
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/ Elona Koni
  • Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy
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  • De Gruyter OnlineGoogle Scholar
/ Nadia Romiti
  • Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy
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  • De Gruyter OnlineGoogle Scholar
/ Aldo Paolicchi
  • Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy
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  • De Gruyter OnlineGoogle Scholar
/ Maria Franzini
  • Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2018-07-23 | DOI: https://doi.org/10.1515/cclm-2018-0412

Abstract

Background

The automated immunochemical serum free light chains (FLC) assays, Freelite (a polyclonal antiserum) and N Latex FLC (a mixture of monoclonal antibodies), are not interchangeable, as they may provide different results on a same sample. This study was aimed to establish if the calibrators contain FLC oligomers, and if different reactivity against monomers and dimers contributes to the discrepancy.

Methods

Gel filtration chromatography fractions of the calibrators were subjected to a Western blot (WB) and analyzed by each reagent. The procedure was repeated after pretreating the N Latex FLC calibrator with the reducing agent dithiothreitol (DTT).

Results

Both calibrators contain FLC dimers and monomers. Both reagents detect (with different sensitivity) FLC kappa monomers and dimers; instead, Freelite detects only FLC lambda dimers, while N Latex FLC detects only FLC monomers. After DTT treatment, only the N Latex lambda still detects FLC with reduced protein thiols, while the reactivity of all other reagents is abolished.

Conclusions

Due to their different reactivity against FLC monomers and oligomers, the Freelite and N Latex FLC are calibrated against different components of their own calibrators, making the two reagents not equivalent. The redox status of FLC determines the immunoreactivity not only of FLC dimers, but also of the monomers.

Keywords: disulfide bonds; free light chains; monoclonal gammopathies; monomers and oligomers

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About the article

Corresponding author: Laura Caponi, MD, PhD, Department of Translational Research and New Technologies in Medicine, Laboratory of Clinical Pathology, University Hospital of Pisa, Building 43, Via Roma, 67, 56126 Pisa, Italy


Received: 2018-04-20

Accepted: 2018-06-25

Published Online: 2018-07-23

Published in Print: 2018-12-19


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

Research funding: Institutional funding of University of Pisa.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization 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 2, Pages 221–229, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-0412.

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