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Detecting alpha-synuclein aggregates with small molecules on single-molecule array.

Jeff Y L Lam1,2,3, Timothy S Chisholm1, Hadia Almahli1

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Chemically synthesized small molecules can now detect alpha-synuclein aggregates, key biomarkers for Parkinson's Disease, using the Single-Molecule Array (SiMoA) assay. This innovation offers a versatile and efficient method for early neurodegenerative disorder diagnosis.

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Area of Science:

  • Biochemistry
  • Neuroscience
  • Analytical Chemistry

Background:

  • Protein aggregates, such as alpha-synuclein, are crucial biomarkers for early diagnosis of neurodegenerative disorders like Parkinson's Disease.
  • The Single-Molecule Array (SiMoA) platform enables ultra-low concentration detection of biomarkers in biofluids.
  • Current SiMoA assays often rely on antibodies for capturing target molecules, which can be labor-intensive to develop.

Purpose of the Study:

  • To develop a next-generation SiMoA assay utilizing chemically synthesized small molecules as capturing agents for alpha-synuclein aggregates.
  • To evaluate the efficacy and specificity of small molecule-based binders compared to traditional antibody-based methods.
  • To enhance the versatility and adaptability of the SiMoA platform for biomarker discovery and diagnostics.

Main Methods:

  • Chemical synthesis of small molecules designed to bind alpha-synuclein aggregates.
  • Functionalization of small molecules with primary amines for conjugation to SiMoA beads.
  • Testing the capture efficiency and specificity of synthesized molecules (e.g., BF-79-2) using recombinant and human blood samples.

Main Results:

  • A novel small molecule, BF-79-2, demonstrated picomolar capture of alpha-synuclein aggregates.
  • The BF-79-2 molecule specifically captured aggregates while excluding monomers.
  • Successful detection of alpha-synuclein aggregates in human blood samples using the small molecule-based SiMoA assay.

Conclusions:

  • Small molecules can effectively replace antibodies as capturing agents in SiMoA assays for detecting protein aggregates.
  • This approach enhances assay versatility, enabling in silico screening and simplified synthesis.
  • The development broadens SiMoA platform capabilities for biomarker discovery and early diagnosis of synucleinopathies.