Molecular Conjugate
AssigneeVentana Medical Systems, Inc.
InventorsChristopher Bieniarz, Julia Ashworth-Sharpe, Casey A. Kernag, Jerome W. Kosmeder, Mark Lefever
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Abstract
A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specific antibody-enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays.
Claims30
A method for forming a conjugate of two or more molecules, comprising:
- reacting a hydrazide thiol linker with a first molecule having a hydrazide-reactive group to form a thiolated first molecule, wherein reacting the hydrazide thiol linker with the first molecule comprises reacting under conditions where a thiol group of the hydrazide thiol linker is substantially present in its neutral acid form;
- and
- reacting the thiolated first molecule with a second molecule, the second molecule having a thiol-reactive group, to form the conjugate.
A conjugate comprising an antibody covalently bonded to a detectable label through a hydrazide thiol linker, wherein the hydrazide thiol linker is one or more of MBH, MBCH, MAMBH, THMBH, BTAL, BTHL, TAGD, THGD, a PEG-based hydrazide-thiol linker, a multifunctional hydrazide thiol linker, a PEG-based multifunctional hydrazide thiol linker, and a polyacrylamide hydrazide linker, wherein a hydrazide group of the linker is covalently bonded to an Fc region of the antibody.
A hydrazide thiol linker having the formula:
- wherein R1 is H or —CONHNH2 and n=1, 2 or 3.
Dependencies are inferred from the claim text.