Ultra-sensitive high performance liquid chromatography–laser-induced fluorescence based proteomics for clinical applications

Patil, Ajeetkumar and Bhat, Sujatha and Pai, Keerthilatha M and Rai, Lavanya and Chidangil, Santhosh (2015) Ultra-sensitive high performance liquid chromatography–laser-induced fluorescence based proteomics for clinical applications. Journal of Proteomics, 127. pp. 202-210. ISSN 1874-3919

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Abstract

An ultra-sensitive high performance liquid chromatography–laser induced fluorescence (HPLC–LIF) based technique has been developed by our group atManipal, 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 profileswere 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. Biological significance : In recent years, proteomics techniques have advanced tremendously in life sciences andmedical sciences for the detection and identification of proteins in body fluids, tissue homogenates and cellular samples to understand biochemicalmechanisms leading to different diseases. Some of themethods include techniques like high performance liquid chromatography, 2D-gel electrophoresis,MALDI–TOF-MS, SELDI–TOF-MS, CE–MS and LC–MS techniques. We have developed an ultra-sensitive high performance liquid chromatography–laser induced fluorescence (HPLC–LIF) based technique, 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 healthy and volunteers with different malignant conditions were recorded by using this set-up. The protein profile data were analyzed using principal component analysis (PCA) for objective classification and detection of malignant, premalignant and healthy conditions. The method is extremely sensitive to detect proteins with limit of detection of the order of femto-moles. The HPLC–LIF combined with PCA as a potential proteomic method for the diagnosis of oral cancer and cervical cancer has been discussed in this paper.

Item Type: Article
Uncontrolled Keywords: HPLC–LIF ; Protein profiling ; Oral cancer ; Cervical cancer.
Subjects: Dentistry > MCODS Manipal > Oral Medicine and Radiology
Departments at MU > Atomic Molecular Physics
Medicine > MMMC Manipal > Microbiology
Medicine > KMC Manipal > Obstetrics & Gynaecology
Depositing User: KMC Library
Date Deposited: 03 Oct 2015 09:36
Last Modified: 03 Oct 2015 09:36
URI: http://eprints.manipal.edu/id/eprint/144299

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