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International Journal of Proteomics
Volume 2012 (2012), Article ID 290457, 8 pages
http://dx.doi.org/10.1155/2012/290457
Research Article

Miniaturized Mass-Spectrometry-Based Analysis System for Fully Automated Examination of Conditioned Cell Culture Media

1Department of Biotechnology, Netherlands Proteomics Centre, Delft University of Technology, Julianalaan 67, 2628BC Delft, The Netherlands
2Institute of Sensor and Actuator Systems, Vienna University of Technology, Gusshausstrasse 27-29/E366, 1040 Vienna, Austria
3Biomedical Research Institute (BJOMED), Hasselt University, Agoralaan building C, 3590 Diepenbeek, Belgium

Received 29 February 2012; Accepted 6 September 2012

Academic Editor: Paul P. Pevsner

Copyright © 2012 Emanuel Weber et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We present a fully automated setup for performing in-line mass spectrometry (MS) analysis of conditioned media in cell cultures, in particular focusing on the peptides therein. The goal is to assess peptides secreted by cells in different culture conditions. The developed system is compatible with MS as analytical technique, as this is one of the most powerful analysis methods for peptide detection and identification. Proof of concept was achieved using the well-known mating-factor signaling in baker’s yeast, Saccharomyces cerevisiae. Our concept system holds 1 mL of cell culture medium and allows maintaining a yeast culture for, at least, 40 hours with continuous supernatant extraction (and medium replenishing). The device’s small dimensions result in reduced costs for reagents and open perspectives towards full integration on-chip. Experimental data that can be obtained are time-resolved peptide profiles in a yeast culture, including information about the appearance of mating-factor-related peptides. We emphasize that the system operates without any manual intervention or pipetting steps, which allows for an improved overall sensitivity compared to non-automated alternatives. MS data confirmed previously reported aspects of the physiology of the yeast-mating process. Moreover, matingfactor breakdown products (as well as evidence for a potentially responsible protease) were found.