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Protein profile study of breast‐tissue homogenates by HPLC‐LIF
Author(s) -
Kumar K. Kalyan,
Chowdary M. V. P.,
Mathew Stanley,
Rao Lakshmi,
Krishna C. Murali,
Kurien Jacob
Publication year - 2009
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.200810046
Subject(s) - chromatography , chemistry , high performance liquid chromatography , mass spectrometry , principal component analysis , proteomics , derivative (finance) , artificial intelligence , biochemistry , computer science , financial economics , economics , gene
Proteomics is a promising approach for molecular understanding of neoplastic processes including response to treatment. Widely used 2D‐gel electrophoresis/Liquid chromatography coupled with mass spectrometry (LC‐MS) are time consuming and not cost effective. We have developed a high‐sensitivity (femto/subfemtomoles of protein/20 μl) High Performance Liquid Chromatography‐Laser Induced Fluorescence HPLC‐LIF instrument for studying protein profiles of biological samples. In this study, we have explored the feasibility of classifying breast tissues by multivariate analysis of chromatographic data. We have analyzed 13 normal, 17 malignant, 5 benign and 4 post‐treatment breast‐tissue homogenates. Data was analyzed by Principal Component Analysis PCA in both unsupervised and supervised modes on derivative and baseline‐corrected chromatograms. Our findings suggest that PCA of derivative chromatograms gives better classification. Thus, the HPLC‐LIF instrument is not only suitable for generation of chromatographic data using femto/subfemto moles of proteins but the data can also be used for objective diagnosis via multivariate analysis. Prospectively, identified fractions can be collected and analyzed by biochemical and/or MS methods. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)