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US 2016/0102332 A1Published application
Methods for making targeted protein toxins by sortase-mediated protein ligation
AssigneePresident and Fellows of Harvard College
InventorsR. John Collier, Andrew J. McCluskey, Bradley L. Pentelute
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Abstract
We described novel methods for making targeted protein toxins by sortase-mediated protein ligation. The methods allow for a toxin and receptor-binding ligand to be ligated under mild conditions in vitro, following their expression and purification as single entities. The methods also provide a much more efficient way of making functional targeted fusion toxins compared to recombinant or chemical production of these structures.
Claims21
A method for making a protein toxin comprising a receptor-binding ligand, the method comprising the steps of:
- a. providing a protein toxin substrate and a C-terminal sortase-recognition motif optionally followed by an affinity epitope that is separated from the toxin by a linker;
- b. providing a targeting moiety comprising an N-terminal peptide;
- and
- c. contacting the protein toxin substrate of step (a) with the targeting moiety of step (b) with a sortase enzyme.
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