Quantification of biomarkers present in physiological samples

Inventors

HRUSOVSKY, Edward Kevin • Wilson, David • Shan, Dandan • Chang, Lei • Song, Linan • Jeromin, Andreas • TOBOS, Carmen Ioana • PATEL, Purvish Prahlad

Assignees

Quanterix Corp

Abstract

The present disclosure relates to immunoassays for NF-L, GFAP, UCH L1, and Tau performed on liquid samples derived from physiological fluids such as venous blood to detect the presence or absence of a physiological condition by quantifying one or a combination of NF-L, GFAP, UCH L1, and Tau at concentrations indicative of the condition.

Core Innovation

The invention relates to assay reagent compositions and a kit for a multiplex, ultra-sensitive digital immunoassay targeting brain-injury and neurodegeneration biomarkers. The kit comprises a plurality of capture agents and a plurality of detection agents for tau protein (Tau) and neurofilament light chain (NF-L), where a first antibody specifically binds Tau and a second antibody specifically binds NF-L in both the capture and detection portions of the assay.

The kit further includes a sample diluent comprising human IgG at a concentration between 1 mg/mL and 10 mg/mL, and a plurality of calibration solutions comprising predetermined concentrations of Tau and NF-L. The calibration solutions enable quantitative determination using the Tau and NF-L concentration sets provided for the assay workflow, and the disclosed embodiments include single-analyte and multi-analyte formats.

Additional disclosed embodiments expand the assay kit to include other neurological biomarker targets such as GFAP and UCH L1 by incorporating additional antibody, capture, and calibration solution components. Further embodiments constrain the Tau species to phosphorylated Tau protein, including phosphorylated Tau 181 (p-Tau-181) and/or phosphorylated Tau 231 (p-Tau-231), and describe antibody presentation using paramagnetic bead conjugates for Tau and NF-L.

Claims Coverage

The independent claim coverage centers on a multiplex immunoassay kit with Tau- and NF-L-specific capture and detection antibodies, a human IgG sample diluent, and Tau/NF-L calibration solutions. Three main inventive features are recited across the independent claim material, with dependent refinements directed to additional biomarker content, phosphorylated Tau species, paramagnetic bead conjugates, immobilization relationships, and heterophile blocking agent requirements.

The claim coverage defines a multiplex Tau/NF-L immunoassay kit with matched Tau- and NF-L-specific capture and detection antibodies, human IgG-containing sample diluent within a specified concentration range, and calibration solutions containing predetermined Tau and NF-L concentrations. The disclosed refinements further address additional neurological biomarkers, phosphorylated Tau species, paramagnetic bead conjugates, immobilization relationships, and heterophile blocking agent requirements.

Stated Advantages

Documented Applications