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Development and validation of a protocol for cell line identification by MALDI-TOF MS

Authors
Journal
BMC Proceedings
1753-6561
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Volume
5
Identifiers
DOI: 10.1186/1753-6561-5-s8-p45
Keywords
  • Meeting Abstract
Disciplines
  • Biology
  • Medicine

Abstract

Development and validation of a protocol for cell line identification by MALDI-TOF MS MEETING ABSTRACT Open Access Development and validation of a protocol for cell line identification by MALDI-TOF MS Guido Vogel1*, André Strauss1, Bernard Jenni1, Dominik Ziegler1, Eric Dumermuth2, Sylvie Antz2, Claudia Bardouille3, Beat Wipf3, Christian Miscenic3, Georg Schmid3, Valentin Pflüger1 From 22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies Vienna, Austria. 15-18 May 2011 Summary Misidentification or cross-contamination of cell lines used in biotechnology or diagnostic settings are a chal- lenge for laboratories and cell culture repositories. Mas- ters et al. [1] among others reported the occurrence of large numbers of unrecognized and unreported misiden- tification or cross-contamination of cell lines. Current methods for the authentication of cell lines such as kar- yotyping, 2D-gel-electrophoresis, restriction fragment length polymorphism (RFLP) or short tandem repeats (STR) are expensive, labor intensive and not routinely applied. In the last decade MALDI-TOF MS became a powerful tool for the rapid and cost effective identifica- tion and taxonomic classification of microorganisms directly from whole cell extracts. We adapted this pro- tein fingerprinting approach for a fast species identifica- tion of eukaryotic cell lines and established as well as validated a reference database. In addition we demonstrate that this new approach has a potential for the rapid characterization of recom- binant protein expression systems. Accurate protein expression and the determination of the molecular mass of the recombinant expressed proteins (20-120kDa) can directly be analyzed from stable transfected and virus infected cultures. Materials and methods 1 ml of fresh culture was transferred into an Eppendorf tube and washed once with 1x PBS. The pellet was resuspended in 70% EtOH for transport and storage at RT. 1µl of the cell pellet was transferred w

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