A transglutamine tag for efficient site-specific bioconjugation
AssigneeMerck Patent GmbH
InventorsBirgit Piater, Ulrich Betz, Harald Kolmar, Vanessa Siegmund
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Abstract
The present invention provides novel peptide sequences for use in microbial transgluatminase-mediated, in particular mTG2-mediated bioconjugations, in particular for the manufacture of antibody-drug-conjugates. Further disclosed are bioconjugation methods employing mTG2 and the novel peptide sequence motifs of the invention. The present invention further provides proteins comprising the novel sequence motifs of the invention as well as polynucleotides encoding the same.
Claims23
Protein comprising at least one acyl glutamine-containing amino donor sequence covalently linked via a y-glutamyl-amide bond to an amino donor-comprising substrate, wherein the at least one acyl glutamine-containing amino acid donor sequence comprises at least the amino acid sequence according to SEQ ID NO:
- 1 (SEQ ID NO:
- 1 (TYFQAYG))
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Protein according to claim wherein
- X1 is any one of A, V, I, L, M or G,
- X2 is one of D, or E,
- X3 is C,
- X4 is C,
- X5, is S, T, or N,
- X6 is one of D, or E.
Protein according to 9, wherein
- X1 is any one of A, V, I, L, M or G,
- X2 is E,
- X3 is C,
- X4 is C,
- X5 is T, and
- X6 is E.
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Method of covalently coupling an amino donor-comprising to an acyl glutamine-containing amino acid donor comprising a polypeptide sequence according to SEQ ID NO:2, comprising the step of:
- contacting said amino donor-comprising substrate and said acyl glutamine-containing amino acid donor in the presence of transglutaminase, preferably mTG2, to obtain a protein according to covalently linked via a γ-glutamyl-amide bond to said amino donor-comprising substrate.
Method according to claim 19, wherein the amino donor-comprising substrate is coupled or covalently bound to dye, drug, ribozyme, nanobody, enzyme, or linker.
21-27. (canceled)
Antibody or antigen-binding fragment thereof, bivalent antibody, or VHH antibody comprising at least one amino acid sequence according to SEQ ID NO:2.
29-40. (canceled)
Dependencies are inferred from the claim text.