Nanospec · Nanopipette-Enabled Mass Spectrometry for Biomarker Discovery in Cerebrospinal Fluid

Neurodegenerative diseases such as Alzheimer’s (AD) and Parkinson’s disease (PD) represent a major global health crisis, with over 9.8 million people in Europe living with dementia. These conditions are driven by intrinsically disordered amyloid proteins, including amyloid-β (Aβ) and α-synuclein (αS), which undergo early conformational changes and aggregate into toxic assemblies. Detecting and characterising these changes at the earliest stages is crucial for timely diagnosis and therapeutic intervention. This fellowship will develop and apply nanopipette-enabled native ion mobility (IM) and top-down mass spectrometry (MS) for the direct analysis of cerebrospinal fluid (CSF), providing the first native MS platform to profile intact proteoforms in complex human biofluids. Nanopipettes are submicron electrospray ionisation emitters that overcome analytical barriers, enabling detection of low-abundance proteins in their native form, mitigating ion suppression and salt adduction. Coupling nanopipette MS with IM and top-down fragmentation will capture conformational ensembles, post translational modifications, and structural fingerprints of disease-associated proteins directly from CSF. Research objectives include: (i) optimising nanopipette workflows with model systems (cell lysates, artificial CSF); (ii) applying these protocols to clinical CSF samples to detect novel disease-specific proteoforms and conformational shifts; and (iii) validating biomarkers using complementary bottom-up proteomics. Collaborative training at UCLA (Prof. Joseph Loo) and VU Amsterdam (Prof. Anouk Rijs) will provide access to cutting-edge Orbitrap, IM-MS, and PhotoSynapt instruments. By pioneering direct native proteoform analysis of CSF, this project addresses an urgent unmet need in biomarker discovery, offering a transformative approach for earlier, more precise AD and PD diagnosis, while establishing a broadly applicable platform for clinical proteomics and next-generation diagnostics.