RESEARCH ARTICLE


One-Step Protein Purification: Use of a Novel Epitope Tag for Highly Efficient Detection and Purification of Recombinant Proteins



Stefan Rasche1, Alexander Martin2, Achim Holzem3, Rainer Fischer1, 3, Helga Schinkel1, Stefan Schillberg*, 1
1 Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Forckenbeckstraße 6, 52074 Aachen, Germany
2 Aachen University of Applied Sciences, Heinrich-Mussmann-Str, 52428 Jülich, Germany
3 RWTH Aachen University, Institute for Molecular Biotechnology, Worringer Weg 1, 52074 Aachen, Germany


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Creative Commons License
© 2011 Rasche et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Forckenbeckstraße 6, 52074 Aachen, Germany; Tel: +49-241-6085-11050; Fax: +49-241-6085-10000; E-mail: stefan.schillberg@ime.fraunhofer.de


Abstract

We used the interaction between an epitope of the Tobacco mosaic virus 54K replicase (tag54) and its corresponding monoclonal antibody 54 (mAb54) for the design of a new epitope-tagging system. We fused the DNA sequence of tag54 and two elongated derivates thereof to the C-terminus of the chloramphenicol acetyltransferase (CAT) gene and produced the tagged proteins in tobacco. Immunoblot and ELISA analysis demonstrated that the tagged proteins were detected with high sensitivity in plant extracts. Moreover, the tag54 system enabled the full recovery of the recombinant CAT with excellent purity through immunoaffinity chromatography.

We consider this novel epitope-tag system to be a valuable tool for both the detection and purification of recombinant proteins.

Keywords: Epitope tag, immunoaffinity, protein purification, tag54, tobacco.