US 11,807,671 B2
Methods for protein ligation and uses thereof
Christopher John Squire, Auckland (NZ); Edward Neill Baker, Auckland (NZ); Paul Gary Young, Auckland (NZ); and Yuliana Yosaatmadja, Auckland (NZ)
Assigned to AUCKLAND UNISERVICES LIMITED, Auckland (NZ)
Appl. No. 16/349,674
Filed by AUCKLAND UNISERVICES LIMITED, Auckland (NZ)
PCT Filed Nov. 16, 2017, PCT No. PCT/NZ2017/050147
§ 371(c)(1), (2) Date May 14, 2019,
PCT Pub. No. WO2018/093274, PCT Pub. Date May 24, 2018.
Claims priority of application No. 726413 (NZ), filed on Nov. 16, 2016.
Prior Publication US 2019/0352343 A1, Nov. 21, 2019
Int. Cl. C07K 14/33 (2006.01); C07K 14/195 (2006.01)
CPC C07K 14/33 (2013.01) [C07K 14/195 (2013.01); C07K 2319/21 (2013.01); C07K 2319/24 (2013.01); C07K 2319/50 (2013.01)] 15 Claims
 
1. A peptide tag and binding partner pair wherein
a) the peptide tag comprises at least about 10 contiguous amino acids of an Ig-like fold of a Cpe-like domain comprising two β-sheets in a β-clasp from a β-clasp containing protein, wherein one of the amino acids is a reactive residue capable of spontaneously forming an intermolecular ester bond in an Ig-like fold of a Cpe-like domain, and wherein the peptide tag does not comprise the entire amino acid sequence of the β-clasp containing protein; and
b) said binding partner comprises a separate fragment of a β-clasp containing protein wherein said fragment comprises at least about 10 contiguous amino acids of a complementary part of an Ig-like fold of a Cpe-like domain from a Cpe-like domain containing protein, wherein one of the amino acids is a reactive residue capable of forming an intermolecular ester bond with the peptide tag;
and wherein either:
i) the reactive residue in (a) is serine or threonine, and the reactive residue in (b) is glutamine or glutamate/glutamic acid; or
ii) the reactive residue in (a) is glutamine or glutamate/glutamic acid, and the reactive residue in (b) is serine or threonine;
and wherein said peptide tag and binding partner are covalently bound to each other only by an intermolecular ester bond between the reactive residues
and wherein the peptide tag or the binding partner or both the peptide tag and the binding partner is covalently linked to one or more heterologous amino acid sequences.