alexa Applying Magnetic Bead Separation / MALDI-TOF Mass Spec
ISSN: 0974-276X

Journal of Proteomics & Bioinformatics
Open Access

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Research Article

Applying Magnetic Bead Separation / MALDI-TOF Mass Spectrometry to Human Tear Fluid Proteome Analysis

Eiichi Sekiyama1, Yumiko Matsuyama2, Daisuke Higo2, Takashi Nirasawa2, Masaya Ikegawa3*, Shigeru Kinoshita1, and Kei Tashiro3

1Kyoto Prefectural University of Medicine, Ophthalmology

2Bruker Daltonics

3Kyoto Prefectural University of Medicine, Genomic Medical Sciences

*Corresponding Author:
Dr. Masaya Ikegawa
Genomic Medical Sciences,
Kyoto Prefectural University of Medicine 465
Kawaramachi Hirokoji, Kamigyo-ku, Kyoto
Japan, Postal code: 602-8566
Tel : +81-75-251-5347,
Fax : +81-75-251-5347,
E-mail: [email protected]

Received Date: August 27, 2008; Accepted Date: October 06, 2008; Published Date: October 10, 2008

Citation: Sekiyama E,Matsuyama Y, Higo D, Nirasawa T, Ikegawa M, et al. (2008) Applying Magnetic Bead Separation / MALDI-TOF Mass Spectrometry to Human Tear Fluid Proteome Analysis. J Proteomics Bioinform 1: 368-373. doi:10.4172/jpb.1000045

Copyright: © 2008 Sekiyama E, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

The proteins and peptides in tears play an important role in preserving the integrity and stability of the ocular surface. Proteomic analysis of tear films will enable us to detect early biological markers of eye diseases, however, it is often hampered by the small amount of tear volume and the low protein concentration. Here we adopted magnetic bead-based purification (ClinProt system) followed by matrix-assisted laser desorption/ionization timeof- flight mass spectrometry (MALDI-TOF-MS) to profile human tear proteins. Basal and reflex tear fluids were collected from normal healthy volunteers using glass microcapillary tubes. Reversed phase (C8) and weak cation exchange (WCX) magnetic beads were applied to obtain multiple components detected as clear signals. Principal component analysis showed a clear differentiation between basal and reflex tears. Among the key alterations, two markedly increased peaks in the reflex tear fluids at m/z 2422.12 and m/z 2721.29 were subsequently analyzed by tandem MS analysis and their source to be proline-rich protein 4 (PRP4). We conclude that magnetic beadbased separation combined with MALDI-TOF-MS (ClinProt MALDI-TOF) appears to be ideally suited for the first-line screening of peptides and proteins in tears.

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