Method and reagents for the detection of an autoantibody
Inventors
MISKE, Ramona • Radzimski, Christiane • Borowski, Kathrin Margalene • DENNO, Yvonne
Assignees
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Publication Number
US-12174186-B2
Publication Date
2024-12-24
Expiration Date
Abstract
A method involves detecting in a sample an autoantibody binding specifically to a mammalian VGLUT. An autoantibody binding specifically to a mammalian VGLUT and a diagnostically useful carrier with a solid phase with an immobilized polypeptide containing a mammalian VGLUT or a variant thereof, are useful. The autoantibody can be used for diagnosing a neurological autoimmune disease or a cancer.
Core Innovation
The invention relates to detecting human autoantibodies that specifically bind mammalian vesicular glutamate transporters (VGLUTs) in a sample. The detected autoantibody specifically binds a mammalian VGLUT, including VGLUT2, and may include VGLUT1 and VGLUT3. The disclosure frames this detection as diagnostically useful for neurological autoimmune disease and/or cancer.
The disclosure provides an approach in which autoantibody binding specificity for VGLUTs is used as the basis for detection in a sample. It identifies VGLUT2 as an autoantigen using immunoprecipitation and mass spectrometry, and reports recombinant immunofluorescence assay using HEK293-VGLUT2 to detect anti-VGLUT2 in sera and CSF from neurological autoimmune disease patients, with clinical associations including paraneoplastic cases.
The disclosure describes assay formats in which VGLUT polypeptides are immobilized on diagnostic carriers, including solid phase and kit formats. Autoantibody binding is detected using immunoassay or immunodetection modalities, including immunofluorescence, ELISA, electrochemiluminescence, radiolabeled immunoassay, and other listed detection modalities.
Specificity is further described through epitope mapping of anti-VGLUT2 to a C-terminal fragment and to residues between aa531-558, and through cross-reactivity patterns across VGLUT isoforms, as well as competitive binding concepts and confirmatory assays. The disclosure also verifies the presence or specificity of autoantibodies against VGLUT1 and VGLUT3 using recombinant immunofluorescence assay, immunoprecipitation, immunoblot, and competitive inhibition.
Claims Coverage
The claim set centers on detecting autoantibodies that specifically bind mammalian vesicular glutamate transporter (VGLUT), with six inventive features in the merged independent and dependent claim coverage.
Detecting autoantibodies specific to mammalian VGLUT
Detecting, in a sample, an autoantibody binding specifically to a mammalian vesicular glutamate transporter (VGLUT).
Carrier-based autoantibody binding and detection workflow
Contacting a carrier with a sample liquid containing an autoantibody that specifically binds mammalian VGLUT, and then contacting the carrier with a detection means to detect the autoantibody binding to VGLUT.
Immobilizing a mammalian VGLUT polypeptide on a carrier
Immobilizing a mammalian VGLUT polypeptide on a carrier.
Immunoassay or immunodetection technique selection
Detecting the autoantibody using one of several listed immunoassay or immunodetection techniques.
Specified immunodetection modalities
Detecting an autoantibody using one of the specified detection methods selected from radiolabeled immunoassay, ELISA, electrochemiluminescence, or indirect immunofluorescence.
Human antibody sample type selection
Using a human sample containing a representative set of antibodies selected from whole blood, plasma, serum, cerebrospinal fluid, or saliva.
Overall, the claim set centers on detecting autoantibodies that specifically bind mammalian VGLUT, with dependent claims further defining a carrier-based workflow, immobilized VGLUT polypeptides, and selection among specified immunoassay or immunodetection modalities using specified human sample types.
Stated Advantages
Provides diagnostically useful detection of autoantibodies specifically binding mammalian VGLUTs in neurological autoimmune disease and/or cancer contexts.
Substantially higher detection than in healthy controls.
Documented Applications
Diagnostic detection of neurological autoimmune disease and/or cancer using VGLUT-specific autoantibodies.
Detection in sera and cerebrospinal fluid.
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