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Novel Approach for Quantitative Measurement of Matrix metalloprotease-1 (MMP1) in Human Breast Cancer Cells Using Mass Spectrometry

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Journal of Analytical Sciences, Methods and Instrumentation 2013年第1期3卷 54-61页

作者： Ming Fa Kathleen Hoch Xuejun Fan William P. Dubinsky Zhiqiang An Ningyan Zhang Proteomics Center Brown Foundation Institute of Molecular Medicine The University of Texas Health Science Center at Houston Houston USA Proteomics Center Brown Foundation Institute of Molecular Medicine The University of Texas Health Science Center at Houston Houston USA. Texas Therapeutics Institute Houston USA

Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.

关键词： Matrix metalloprotease-1 (MMP1) SKBR3 Breast Cancer Cells Mass Spectrometry Selected Reaction Monitoring Absolute Quantification

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