Protein profile study of breast‐tissue homogenates by HPLC‐LIF
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 mul) 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.
- SourceAvailable from: Atakan Ekiz[Show abstract] [Hide abstract]
ABSTRACT: Proteins are important targets in cancer research because malignancy is associated with defects in cell protein machinery. Protein profiling is an emerging independent subspecialty of proteomics that is rapidly expanding and providing unprecedented insight into biological events. Quantitative assessment of protein levels in hematologic malignancies seeks a comprehensive understanding of leukemia-associated protein patterns for use in aiding diagnosis, follow-up treatment, and the prediction of clinical outcomes. Many recently developed high-throughput proteomic methods can be applied to protein profiling. Herein the importance of protein profiling, its exploitation in leukemia research, and its clinical usefulness in the treatment and diagnosis of various cancer types, and techniques for determining changes in protein profiling are reviewed.Turkish Journal of Haematology 03/2011; DOI:10.5152/tjh.2011.01 · 0.34 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: An ultra-sensitive high performance liquid chromatography–laser induced fluorescence (HPLC–LIF) based technique has been developed by our group at Manipal, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from volunteers (normal, and different pre-malignant/malignant conditions) were recorded using this set-up. The protein profiles were analyzed using principal component analysis (PCA) to achieve objective detection and classification of malignant, premalignant and healthy conditions with high sensitivity and specificity. The HPLC–LIF protein profiling combined with PCA, as a routine method for screening, diagnosis, and staging of cervical cancer and oral cancer, is discussed in this paper.Journal of Proteomics 05/2015; 13. DOI:10.1016/j.jprot.2015.05.006 · 3.93 Impact Factor