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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 / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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1437-4331
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Volume 48, Issue 2 (Feb 2010)

Issues

Soluble mesothelin related peptides (SMRP) and osteopontin as protein biomarkers for malignant mesothelioma: analytical validation of ELISA based assays and characterization at mRNA and protein levels

Alex J. Rai
  • Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, USA
/ Raja M. Flores
  • Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Anu Mathew
  • Department of Clinical Laboratories, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Rita Gonzalez-Espinoza
  • Department of Clinical Laboratories, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Matthew Bott
  • Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Marc Ladanyi
  • Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Valerie Rusch
  • Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
/ Martin Fleisher
  • Department of Clinical Laboratories, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Abstract

Background: There is a need to identify reliable markers for malignant mesothelioma. Soluble mesothelin related peptides (SMRP) and osteopontin (OPN) have gained interest in recent years for this purpose.

Methods: SMRP (Fujirebio Diagnostics Inc.) and OPN (R&D Inc.) ELISA methods were evaluated for multiple parameters. Concentrations were measured in blood from patients with mesothelioma, normal healthy volunteers, and patients with other (non-mesothelioma) cancers. In silico analysis was performed using the GeoProfiles database. At the protein level, SMRP and OPN were measured in cell culture supernatants, and values were compared in patients pre- and post-extrapleural pneumonectomy.

Results: The SMRP assay demonstrates intra-assay CVs of 2.3% and 3% (at 4.6 nM and 13.7 nM, respectively), and inter-assay CVs of 3.5% and 3.7% at the same concentrations. The limit of detection (LOD) is 0.182 nM. The OPN assay demonstrates intra-assay CVs of 5.8%, 4.1%, and 5.2% (at 1.9, 5.1, and 11.1 ng/mL, respectively), and inter-assay CVs of 8.5%, 8.4%, and 12.1% at the same concentrations. The LOD is 0.032 ng/mL. Both SMRP and OPN in mesothelioma patients were significantly higher than in patients with other (non-mesothelioma) cancer and in healthy volunteers. The two markers do not appear to correlate with each other and exhibit different tissue expression patterns. Protein concentrations of these markers are highest in different sets of cell lines. Finally, SMRP but not OPN concentrations were decreased in five of seven consecutive patients after extrapleural pneumonectomy, compared to their respective pre-operative values.

Conclusions: These assays provide reliable and reproducible quantitation of SMRP and OPN proteins. Both are increased in mesothelioma patients compared to non-mesothelioma controls. However, the two analytes do not correlate with each other and show distinct expression profiles and protein expression. Concentrations of SMRP but not OPN are decreased in post-surgical samples. Our results further characterize these markers, establish assay performance characteristics, and lay the groundwork for further studies to measure these markers in blood.

Clin Chem Lab Med 2010;48:271–8.

Keywords: analytical validation; cancer biomarkers; mesothelin; mesothelioma; osteopontin; soluble mesothelin related peptides

About the article

Corresponding author: Alex J. Rai, PhD, DABCC, FACB, Assistant Professor of Pathology, Director, Special Chemistry Laboratory, Chief Scientific Officer, Center for Advanced Laboratory Medicine, Columbia University Medical Center 622 West 168th Street, CHONY 2C-224, New York, NY 10032, USA Fax: +1-212-305-7553,


Received: 2009-02-24

Accepted: 2009-11-01

Published in Print: 2010-02-01


Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2010.066. Export Citation

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