Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-like Polypeptides

Authors


  • We thank Wafa Hassouneh for performing light scattering data collection and analysis and Kate Clancy for assistance with determining the initial reaction conditions. This work was supported by a University Scholars Fellowship awarded by the Graduate School at Duke University and by the National Institutes of Health through grants 5T32 GM008487, R01 GM061232, and R01 AI46611.

Abstract

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Sorting it out: Applied in tandem, elastin-like polypeptides (ELPs, see scheme) and the Sortase A (SrtA) transpeptidase provide a method for chromatography-free purification of recombinant proteins and optional, site-specific conjugation of the protein to a small molecule (yellow). This system provides an efficient mechanism for generating bioactive proteins at high yields and purities.

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