Antibodies, kit and method for detecting amyloid beta oligomers
Inventors
Goure, William F. • Gaspar, Renee C. • McCampbell, Alexander • Savage, Mary J. • Shughrue, Paul J. • Wang, Fubao • Wang, Weirong • Wolfe, Abigail L. • Zhang, Ningyan • Zhao, Wei-Qin • Xu, Min
Assignees
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Publication Number
US-9310383-B2
Publication Date
2016-04-12
Expiration Date
Abstract
This invention is a selective Aβ oligomer kit and immunoassay method capable of reliably and sensitively detecting Aβ oligomers in a biological sample of a patient. In one embodiment the inventive assay uses a pair of anti-Aβ oligomer antibodies, as capture and detection antibodies, to detect and quantify Aβ oligomers. The method can be used to differentiate Alzheimer's disease (AD) patients from non-AD patients and/or to stratify AD patients according to the severity of their disease.
Core Innovation
The invention relates to an Aβ oligomer-selective immunoassay kit and method for reliably detecting and quantifying amyloid beta (Aβ) oligomers. The approach uses a capture antibody and a detection antibody that both recognize N-terminal linear epitopes of the amyloid beta 1-42 peptide, including epitopes associated with residues 1–20. The capture antibody provides conformational and oligomer preference relative to Aβ monomers and other Aβ forms.
The method includes obtaining a biological sample having oligomers of amyloid beta, contacting the sample with an isolated capture antibody defined by specific light- and heavy-chain variable region CDR sequences, and forming a capture antibody/oligomer complex. The complex is then contacted with a detection antibody selected from a panel of named antibodies, where the detection antibody recognizes an N-terminal linear epitope of amyloid beta 1-42 peptide, and the resulting complex is detected.
Documented assay performance includes improved limits of detection and selectivity using paramagnetic microparticle-based detection. The document further describes clinical/biological results showing higher Aβ oligomer levels in clinically confirmed Alzheimer’s disease versus controls and correlation with cognitive performance (MMSE), including pattern differentiation of Aβ monomer species.
Claims Coverage
The partial content provides two independent claims. Collectively, they cover a kit comprising an antibody with defined variable-region CDR sequences and a detectably labeled antibody selected from a specified group, and a method for detecting amyloid beta oligomers by forming a capture antibody/oligomer complex, contacting with a detection antibody that recognizes an N-terminal linear epitope of amyloid beta 1-42, and detecting the complex.
CDR-defined capture antibody within an immunoassay kit
A kit comprising an isolated antibody or antigen binding fragment having specified light- and heavy-chain variable region CDR1/2/3 sequence patterns with constrained Xaa residue options, as defined by the SEQ ID NOs.
Detectably labeled antibody selected from a defined panel
The kit further includes an antibody selected from the group consisting of 6E10, BAM-10, W0-2, 26D6, 2A10, 2B4, 4C2, 4E2, 2H4, 20C2, 2D6, 5F10, 1F4, 1F6, 2E12, 3B3 or 82E1, wherein the selected antibody is detectably labeled.
Capture antibody/oligomer complex with CDR-defined binding
A method for detecting oligomers of amyloid beta by contacting a biological sample containing Aβ oligomers with an isolated antibody or antigen binding fragment having the specified light- and heavy-chain variable region CDR1/2/3 sequence patterns, under conditions sufficient to form a capture antibody/oligomer of amyloid beta complex.
Detection antibody recognizing N-terminal linear epitope of Aβ1-42
Contacting the capture antibody/oligomer complex with a detection antibody selected from the group consisting of 6E10, BAM-10, W0-2, 26D6, 2A10, 2B4, 4C2, 4E2, 2H4, 20C2, 2D6, 5F10, 1F4, 1F6, 2E12, 3B3 or 82E1 under conditions sufficient to form a capture antibody/oligomer of amyloid beta/detection antibody complex, wherein the detection antibody recognizes an N-terminal linear epitope of amyloid beta 1-42 peptide.
The claim set provides coverage for an immunoassay kit and method based on a CDR-defined capture antibody that forms a capture antibody/amyloid beta oligomer complex, followed by binding and detection using a detectably labeled detection antibody selected from a specified panel that recognizes an N-terminal linear epitope of amyloid beta 1-42.
Stated Advantages
Improved limits of detection and selectivity for detecting Aβ oligomers, including reported selectivity ratios.
Differentiation of clinically confirmed Alzheimer’s disease from controls using CSF Aβ oligomer levels and correlation with MMSE cognitive performance.
Differentiation of Aβ monomer patterns, including reduced Aβ1-42 monomer with unchanged Aβ1-40.
Improved stability upon storage.
Documented Applications
Detection and quantification of amyloid beta oligomers in a biological sample, including cerebrospinal fluid (CSF).
Clinical/biological analysis comparing Aβ oligomer levels in clinically confirmed Alzheimer’s disease versus controls, with correlation to MMSE cognitive performance.
Development/description of assay concepts including pharmacodynamic (PD) and target engagement (TE) assay approaches for therapeutic antibody monitoring.
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