Label-Free Acoustic Wave Detection of Myelin Basic Protein for Monitoring of Neurodegenerative Disease.
Spagnolo Sandro S, Mikuličová Sára S, Davoudian Katharina K, Sontakke Kiran K et al.
Myelin basic protein (MBP) is released into cerebrospinal fluid during demyelination and is therefore a promising biomarker for the detection of neurodegenerative diseases such as multiple sclerosis. For the label-free detection of MBP, we developed an antifouling aptasensor based on thickness shear mode sensing. DNA aptamers were immobilized on gold-coated piezoelectric crystals functionalized with an engineered antifouling self-assembled monolayer based on 3-(2-mercaptoethoxy)propanoic acid (HS-MEG-COOH) combined with either 2-(2-mercaptoethoxy)ethanol (HS-MEG-OH) or dithiothreitol (DTT) as spacers. The binding of recombinant human MBP to the aptamer was confirmed with a corresponding decrease in resonance frequency in both phosphate-buffered saline (PBS) and artificial cerebrospinal fluid (aCSF). The aptasensor achieved a limit of detection of 11 ng mL-1 in PBS and 2 ng mL-1 in aCSF. PBS measurements were primarily applied to determine the aptasensor functionality, whereas experiments in aCSF evaluated MBP detection at concentrations relevant to reported levels in biological CSF. Selectivity was assessed with aptamer-free surfaces, cytochrome c, and human serum albumin (HSA) with much lower responses, while concentration-dependent MBP detection was maintained in aCSF despite the increased complexity of the matrix. The antifouling performance of the modified sensing surface was further demonstrated in aCSF by the significantly reduced frequency and dissipation responses compared to bare gold. With future validation in human cerebrospinal fluid (CSF) samples, the developed biosensor has the potential to enable sensitive, label-free detection of MBP in CSF for the identification and monitoring of neurodegenerative diseases.