Homogenous antibody drug conjugates via enzymatic methods
First Claim
1. A method of making an antibody drug conjugate comprising an antibody specifically conjugated to a conjugate moiety comprising:
- contacting an antibody composition comprising the antibody with the conjugate moiety in the presence of a transglutaminase under a condition sufficient to generate the antibody drug conjugate, wherein at least about 50% of the antibody in the antibody composition is glycosylated in the Fc-region, wherein the conjugate moiety is conjugated to an endogenous acceptor glutamine residue on the antibody, and wherein the transglutaminase is a wildtype transglutaminase and the concentration of the transglutaminase in the reaction mixture is about 0.1 mg/ml to about 1 mg/ml.
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Abstract
The present application in one aspect provide Fc-containing polypeptide conjugates comprising an Fc-containing polypeptide conjugated to a conjugate moiety, wherein the Fc-containing polypeptide comprises an N-glycosylated Fc region comprising an acceptor glutamine residue flanked by an N-glycosylation site and wherein the conjugate moiety is conjugated to the Fc-containing polypeptide via the acceptor glutamine residue. Also provided are methods of making such Fc-containing polypeptide conjugates by using a wildtype or engineered transglutaminases. Further provided are engineered transglutaminases specifically designed for carrying out such reactions.
16 Citations
16 Claims
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1. A method of making an antibody drug conjugate comprising an antibody specifically conjugated to a conjugate moiety comprising:
- contacting an antibody composition comprising the antibody with the conjugate moiety in the presence of a transglutaminase under a condition sufficient to generate the antibody drug conjugate, wherein at least about 50% of the antibody in the antibody composition is glycosylated in the Fc-region, wherein the conjugate moiety is conjugated to an endogenous acceptor glutamine residue on the antibody, and wherein the transglutaminase is a wildtype transglutaminase and the concentration of the transglutaminase in the reaction mixture is about 0.1 mg/ml to about 1 mg/ml.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method of making an antibody drug conjugate comprising an antibody specifically conjugated to a conjugate moiety comprising a small molecule handle and an active moiety comprising:
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a) contacting an antibody composition comprising the antibody with the small molecule handle in the presence of a transglutaminase under a condition sufficient to generate an intermediate conjugate comprising the antibody specifically conjugated to the small molecule handle, wherein the transglutaminase is a wildtype transglutaminase and the concentration of the transglutaminase in the reaction mixture is about 0.1 mg/ml to about 1 mg/ml, and b) contacting the intermediate conjugate with the active moiety thereby obtaining the antibody drug conjugate, wherein at least about 50% of the antibody in the antibody composition is glycosylated in the Fc-region, wherein the conjugate moiety is conjugated to an endogenous acceptor glutamine residue on the antibody, and wherein the active moiety is selected from the group consisting of a moiety that improves the pharmacokinetic property of the antibody composition, a therapeutic moiety, and a diagnostic moiety. - View Dependent Claims (15)
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16. A method of making an antibody drug conjugate comprising an antibody specifically conjugated to a conjugate moiety comprising:
- contacting an antibody composition comprising the antibody with the conjugate moiety in the presence of a transglutaminase under a condition sufficient to generate the antibody drug conjugate, wherein at least about 50% of the antibody in the antibody composition is glycosylated in the Fc-region, wherein the conjugate moiety is conjugated to an endogenous acceptor glutamine residue on the antibody, and wherein the transglutaminase is a wildtype transglutaminase with an added proline residue at the N-terminus, and optionally a purification tag, and the concentration of the transglutaminase in the reaction mixture is about 0.1 mg/ml to about 1 mg/ml.
Specification