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Covalent Attachment of Proteins to Solid Supports and Surfaces via Sortase-Mediated Ligation

Chan, L, Cross, HF, She, JK, Cavalli, G, Martins, HFP and Neylon, C (2007) Covalent Attachment of Proteins to Solid Supports and Surfaces via Sortase-Mediated Ligation PLOS ONE, 2 (11). ? - ?. ISSN 1932-6203

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Abstract

Background. There is growing interest in the attachment of proteins to solid supports for the development of supported catalysts, affinity matrices, and micro devices as well as for the development of planar and bead based protein arrays for multiplexed assays of protein concentration, interactions, and activity. A critical requirement for these applications is the generation of a stable linkage between the solid support and the immobilized, but still functional, protein. Methodology. Solid supports including crosslinked polymer beads, beaded agarose, and planar glass surfaces, were modified to present an oligoglycine motif to solution. A range of proteins were ligated to the various surfaces using the Sortase A enzyme of S. aureus. Reactions were carried out in aqueous buffer conditions at room temperature for times between one and twelve hours. Conclusions. The Sortase A transpeptidase of S. aureus provides a general, robust, and gentle approach to the selective covalent immobilization of proteins on three very different solid supports. The proteins remain functional and accessible to solution. Sortase mediated ligation is therefore a straightforward methodology for the preparation of solid supported enzymes and bead based assays, as well as the modification of planar surfaces for microanalytical devices and protein arrays.

Item Type: Article
Additional Information: Copyright: 2007 Chan 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.
Uncontrolled Keywords: Science & Technology, Multidisciplinary Sciences, Science & Technology - Other Topics
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Divisions: Faculty of Engineering and Physical Sciences > Chemistry
Depositing User: Melanie Hughes
Date Deposited: 09 Nov 2010 10:41
Last Modified: 23 Sep 2013 18:39
URI: http://epubs.surrey.ac.uk/id/eprint/2605

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