Protein profile study of breast‐tissue homogenates by HPLC‐LIF |
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Authors: | K Kalyan Kumar M V P Chowdary Stanley Mathew Lakshmi Rao C Murali Krishna Jacob Kurien |
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Institution: | 1. Center for Atomic and Molecular Physics, Manipal University, Manipal, 576 104, Karnataka, India;2. Department of Surgical Oncology, Shirdi Sai Baba Cancer Hospital, Manipal University, Manipal, 576 104, Karnataka, India;3. Department of General Surgery, Kasturba Hospital, Manipal University, Manipal, 576 104, Karnataka, India;4. Department of Pathology, Kasturba Medical College, Manipal University, Manipal, 576 104, Karnataka, India;5. Present adress: Scientific Officer ‘E’ and Principal Investigator, Chilakapati Laboratory, ACTREC, TMC, Kharghar, Sector ‘22’, Navi Mumbai – 410 208, India |
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Abstract: | 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) |
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Keywords: | chromatogram HPLC‐LIF breast cancer PCA |
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