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
Article Information
Identifiers and Pagination:
Year: 2011Volume: 5
First Page: 1
Last Page: 6
Publisher ID: TOBIOTJ-5-1
DOI: 10.2174/1874070701105010001
Article History:
Received Date: 30/12/2010Revision Received Date: 10/03/2011
Acceptance Date: 14/03/2011
Electronic publication date: 5/5/2011
Collection year: 2011
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.
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.