1：Experimental Design and Method Selection:

Designed a hybrid PEF–AIE sensor using plasmonic Au@SiO2 nanoparticles coupled with AIE molecules to monitor protein conformational changes. Theoretical models include aggregation-induced emission and plasmon-enhanced fluorescence effects.

2：Sample Selection and Data Sources:

Used prion protein (PrP) as a model, with cellular prion protein (PrPC) and disease-associated prion protein (PrPSc) obtained from Calbiochem. Human serum samples were purchased from Biodee.

3：List of Experimental Equipment and Materials:

Synthesized three anthracene derivatives (BMNVA, BHNVA, BSNVA), used Au nanocubes coated with silica (Au@SiO2), aptamer for PrP, and various chemicals from suppliers like Alfa Aesar, Tokyo Chemical Industry, Sigma-Aldrich. Instruments include JEOL 400 MHz spectrometer, Thermo Fisher Scientific LTQ-XL mass spectrometer, Shimadzu UV-2450 spectrometer, RF-6000 fluorescence spectrometer, JEOL 2010 TEM, Hitachi S-8010 SEM, Malvern Nano-ZS Zetasizer, Nikon ALR confocal microscope, JASCO J-810 spectropolarimeter.

4：Experimental Procedures and Operational Workflow:

Synthesized AIE molecules via Witting–Horner reaction, fabricated Au@SiO2 nanoparticles, modified with aptamer, mixed with PrP samples and BSNVA, and measured fluorescence changes. For conformational monitoring, PrPC was incubated at pH

5：0 and 65°C, aliquots taken at intervals for fluorescence measurement. Data Analysis Methods:

Fluorescence intensity changes were analyzed to monitor conformational kinetics. Molecular docking simulations using AutoDock 4.2 with Lamarckian Genetic algorithm to study interactions between BSNVA and PrP structures. CD spectroscopy and SEM for validation.