UCLA researchers from the Department of Neurology have developed a novel duplex assay for the detection of α-synuclein and pS129-α-synuclein proteins in biological samples.
BACKGROUND: The accumulation of a-synuclein (a-syn) into protein aggregates is a defining feature of neurodegenerative disorders known as synucleinopathies, including Parkinson’s disease (PD), dementia with Lewy bodies, and and multiple system atrophy (MSA). Phosphorylation of α-synuclein at serine 129 (pS129-a-syn) accounts for ~4% of total α-synuclein in the healthy brain but rises up to ~90% within pathological aggregates. While pS129-α-syn can be readily visualized by immunohistochemistry, accurately quantifying it in tissue extracts or bodily fluids is challenging because of the high sensitivity required. Existing colorimetric Enzyme Linked Immunosorbent Assays (ELISAs) provide insufficient detection sensitivity to meet this need, highlighting a clinical demand for a high-throughput, ultrasensitive assay to measure a-syn and pS129–α-syn to enable earlier diagnosis and improved monitoring and treatment of synucleinopathies.
INNOVATION: Researchers at UCLA have developed a duplex electrochemiluminescence immunoassay (ECLIA), based on Meso Scale Discovery (MSD) technology, that enables high-sensitivity measurement of pS129-a-syn in biological samples. UCLA researchers optimized antibody configurations and incubation parameters to identify a combination with (i) exceptional specificity, (ii) broad dynamic linear range, and (iii) high reproducibility, which were all validated through quantifiable ECLIA signal. The antibody assay was then applied to biological samples, where increased signal was observed in brain tissue from mice with elevated α-syn levels as well as in samples from patients with PD or MSA. In summary, UCLA researchers have generated a novel duplex ECLIA that sensitively and specifically detects total α-synuclein and pS129-α-synuclein simultaneously to enable better detection of synucleinopathies. This platform provides a powerful tool for offering earlier therapeutic intervention and characterization of synucleinopathies.
POTENTIAL APPLICATIONS:
- Detection of pS129-a-syn in patients with synucleinopathies
- Application of this assay for detection of other low abundance proteins found in diseased states
ADVANTAGES:
- High sensitivity to measure low abundance proteins, a-syn and pS129-a-syn, which are abundant in the brain of patients with synucleinopathies but exist at low concentrations in peripheral biofluids.
- The assay is relatively affordable.
- The nature of the duplex assay allows for substantial saving in precious sample volume,.
- Currently, only singlex assays exist in the market for total a-syn and some commercial ELISA assays with lower sensitivity for pS129-a-syn.
DEVELOPMENT-TO-DATE: UCLA researchers have developed an innovative duplex assay to detect total α-synuclein and pS129-α-synuclein of human samples in an in vitro laboratory setting.
Related Papers (from the inventors only):
Dutta et al., ACS Chemical Neuroscience 2023 14 (7), 1238-1248 DOI: 10.1021/acschemneuro.2c00676
Keywords: Duplex assay, α-synuclein, pS129-α-synuclein, synucleinopathies, plasma, serum, CSF, extracellular vesicles, ELISA, MSD, ECLIA