Engineered polypeptide conjugates and methods for making thereof using transglutaminase
The present invention provides engineered polypeptide conjugates (e.g., antibody-drug-conjugates, toxin-(biocompatible polymer) conjugates, antibody-(biocompatible polymer) conjugates, and bispecific antibodies) comprising acyl donor glutamine-containing tags and amine donor agents. In one aspect, the invention provides an engineered Fc-containing polypeptide conjugate comprising the formula (Fc-containing polypeptide)-T-A, wherein T is an acyl donor glutamine-containing tag engineered at a specific site or comprises an endogenous glutamine made reactive by the Fc-containing polypeptide engineering, wherein A is an amine donor agent, and wherein the amine donor agent is site-specifically conjugated to the acyl donor glutamine-containing tag or the endogenous glutamine. The invention also provides methods of making engineered polypeptide conjugates using transglutaminase.
1. An isolated engineered Fc-containing antibody conjugate comprising the formula: (Fc-containing antibody)-T-A, wherein T is an acyl donor glutamine-containing tag engineered at a specific site; wherein A is an amine donor agent; wherein the amine donor agent is site-specifically conjugated to the acyl donor glutamine-containing tag at position 294 or 296 (EU numbering system) in the Fc-containing antibody, wherein the acyl donor glutamine-containing tag is LLQG (SEQ ID NO:3).
2. The engineered Fc-containing antibody conjugate of claim 1 , wherein the antibody is an IgG.
3. The engineered Fc-containing antibody conjugate of claim 1 , wherein the amine donor agent comprises the formula:
X-Y-Z, wherein X is an amine donor unit; Y is a linker; and Z is an agent moiety.
4. The engineered Fc-containing antibody conjugate of claim 3 , wherein the amine donor unit-linker (X-Y) is selected from the group consisting of Ac-Lys-Gly, aminocaproic acid, Ac-Lys-β-Ala, amino-PEG2-C2, amino-PEG3-C2, amino-PEG6-C2, Ac-Lys-Val-Cit-PABC, aminocaproyl-Val-Cit-PABC, putrescine, and Ac-Lys-putrescine.
5. The engineered Fc-containing antibody conjugate of claim 3 , wherein the agent moiety is a cytotoxic agent.
6. The engineered Fc-containing antibody conjugate of claim 5 , wherein the cytotoxic agent is selected from the group consisting of an anthracycline, an auristatin, a dolastatin, a duocarmycin, an enediyne, a geldanamycin, a maytansine, a puromycin, a taxane, a vinca alkaloid, SN-38, a tubulysin, a hemiasterlin, and stereoisomers, and isosteres thereof.