Protein chip based miniaturized assay for the simultaneous quantitative monitoring of cancer biomarkers in tissue extracts.

Abstract

A multiplexed fluorescence immunoassay using a novel planar waveguide technology-based microarray system, ZeptoMARK (Zeptosens), was developed to detect simultaneously urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), and vascular endothelial growth factor (VEGF) in extracts of breast cancer tissues. The three analytes assay was cross-validated with single-analyte ELISA/chemiluminescence immunosorbent assay tests, revealing good correlations and enhanced assay sensitivities (LODs) of 1 pg/mL for uPA, 33 pg/mL for PAI-1, and 1 pg/mL for VEGF. Values were well within the 80-120% limits for assay recovery and within the +/-20% limits for assay precision. The uPA, PAI-1, and VEGF results obtained from 50 breast cancer cytosols using the protein array system demonstrated that the microarray-based multiplexed assay is a sensitive and robust tool to be used for the simultaneous quantification of cancer markers in small breast cancer tissue samples (core biopsies). The miniaturized, multiplexed assay format has a potential to be used for the quantitative analysis of a larger set of validated markers with significance in disease management.

• Authors: Ulrike Weissenstein, Michael J Schneider, Michael Pawlak, Jonas Cicenas, Serenella Eppenberger-Castori, Peter Oroszlan, Sabine Ehret, Anneke Geurts-Moespot, Fred C G J Sweep, Urs Eppenberger
• Journal: Proteomics (Proteomics) Vol. 6 Issue 5 Pg. 1427-36 (Mar 2006) ISSN: 1615-9853 [Print] Germany
• PMID: 16440370 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Biomarkers, Tumor
• Plasminogen Activator Inhibitor 1
• Tissue Extracts
• Vascular Endothelial Growth Factor A
• Urokinase-Type Plasminogen Activator

Topics

• Biomarkers, Tumor (analysis)
• Breast Neoplasms (chemistry)
• Female
• Humans
• Image Processing, Computer-Assisted
• Immunoassay (methods)
• Plasminogen Activator Inhibitor 1 (analysis)
• Protein Array Analysis (instrumentation, methods)
• Reproducibility of Results
• Sensitivity and Specificity
• Statistics as Topic
• Tissue Extracts (chemistry)
• Urokinase-Type Plasminogen Activator (analysis)
• Vascular Endothelial Growth Factor A (analysis)