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  • Title: Efficient sortase-mediated N-terminal labeling of TEV protease cleaved recombinant proteins.
    Author: Sarpong K, Bose R.
    Journal: Anal Biochem; 2017 Mar 15; 521():55-58. PubMed ID: 28088451.
    Abstract:
    A major challenge in attaching fluorophores or other handles to proteins is the availability of a site-specific labeling strategy that provides stoichiometric modification without compromising protein integrity. We developed a simple approach that combines TEV protease cleavage, sortase modification and affinity purification to N-terminally label proteins. To achieve stoichiometrically-labeled protein, we included a short affinity tag in the fluorophore-containing peptide for post-labeling purification of the modified protein. This strategy can be easily applied to any recombinant protein with a TEV site and we demonstrate this on Epidermal Growth Factor Receptor (EGFR) and Membrane Scaffold Protein (MSP) constructs.
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