US20260219283
2026-07-30
Physics
G01N33/6896
The patent application discusses a novel method for examining α-synuclein, a protein associated with neurodegenerative diseases such as Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. This method addresses the need for a sensitive assay to measure phosphorylated α-synuclein at serine 129 (pS129-α-syn), which is significantly elevated in these conditions. The technology utilizes electrochemiluminescence (ECL) to achieve high sensitivity, allowing detection of pS129-α-syn in very low concentrations, crucial for biomarker studies.
α-Synuclein aggregation is a hallmark of synucleinopathies, and pS129-α-syn is a promising biomarker due to its elevated presence in pathological conditions. Traditional methods to measure this phosphorylated protein face challenges in sensitivity and specificity, often due to variability in antibodies and difficulties in generating standard curves. Existing color-based ELISAs lack the sensitivity required for certain biomarker studies, necessitating the development of more advanced assays.
The invention introduces a duplex assay that measures both total and phosphorylated α-synuclein in the same sample, optimizing sample volume usage. Initial attempts faced challenges with reduced signal for pS129-α-syn, which were overcome by adjusting antibody concentrations and extending incubation times. This led to significant improvements in assay sensitivity, allowing for the detection of pS129-α-syn at picogram per milliliter levels in biological samples.
The method involves attaching a pS129-α-syn capture antibody to a matrix, introducing a sample, and using a detection antibody with an electrochemiluminescent label. Light emitted from this label, when electricity is applied, correlates to pS129-α-syn concentration. This approach enables ultra-sensitive detection, with the ability to measure pS129-α-syn at concentrations as low as 1-9 μg/mL. The method also includes generating a standard curve using a stable pS129-α-syn standard.
The invention's duplex assays utilize different capture and detection antibodies, optimizing their concentration ratios for sensitivity. This allows for accurate measurement of both total and phosphorylated α-synuclein in small sample volumes. The assay is applicable to various biological samples, including cerebral spinal fluid and blood-derived exosomes, offering potential for use in diagnostic biomarker development, disease monitoring, and clinical trials.