Epitope imprinting of alpha-synuclein for sensing in Parkinson's brain organoid culture medium

Parkinson's disease (PD) is a progressive nervous system disorder that affects movement, whose early signs may be mild and unnoticed. alpha-Synuclein has been identified as the major component of Lewy bodies and Lewy neurites, which are the characteristic proteinaceous deposits that are the hallmarks of PD. In this work, three alpha-synuclein peptides were synthesized as templates for the molecular imprinting of conductive polymers to enable recognition of alpha-synuclein via ultrasensitive electrochemical measurements. The peptide sequences encompassed specific residues where mutations are known to accelerate PD (though the target sequences, in this study, were wild-type.) The different peptide targets were all successfully imprinted, but with differing imprinting effectiveness, probably owing to differences in target carboxylic acids (which can bind to the aniline (AN) m-aminobenzenesulfonic acid (MSAN) MIP polymers.) Composition of the imprinted polymer, (the mole proportions of AN and MSAN), and the concentrations and sequences of imprinted peptide templates were optimized by measuring the electrochemical responses to target peptides. The imprinted electrode can detect alpha-synuclein at fg/mL levels, and was therefore used to measure alpha-synuclein in the culture medium of human brain organoids generated from normal and idiopathic PD patients.

Automated summary

In this study, α-synuclein peptides were synthesized and used as templates for molecular imprinting of conductive polymers to recognize α-synuclein. By optimizing the composition of the imprinted polymer and the concentrations and sequences of imprinted peptides, the sensitivity of detection was significantly improved. This technology can be applied to measure α-synuclein levels in brain organoids from Parkinson's disease patients.