Label-free high-throughput screening assay for inhibitors of Alzheimer's amyloid-β peptide aggregation based on MALDI MS.
Aggregation of amyloid-β (Aβ) peptides is causatively linked to Alzheimer's disease (AD); thus, suppression of this process by small molecule inhibitors is a widely accepted therapeutic and preventive strategy for AD. Screening of the inhibitors of Aβ aggregation deserves much attention; however, despite intensive efforts, there are only a few high-throughput screening methods available, all of them having drawbacks related to the application of external fluorescent probes or artificial Aβ derivatives. We have developed a label-free MALDI MS-based screening test for inhibitors of Aβ₄₂ fibrillization that exhibits high sensitivity, speed, and automation possibilities suitable for high-throughput screening. The test was evaluated by transmission electron microscopy and compared with a fluorimetric thioflavin-based assay, where interference of a number of tested compounds with thioflavin T binding and/or fluorescence caused false-positive results. The MALDI MS-based method can significantly speed up in vitro screening of compound libraries for inhibitors of Aβ₄₂ fibrillization.
Zovo
Kairit
K
Department of Gene Technology, Tallinn University of Technology, Akadeemia tee 15, Tallinn 12618, Estonia.
Helk
Eneken
E
Karafin
Ann
A
Tõugu
Vello
V
Palumaa
Peep
P
D016229:Amyloid beta-Peptides / Q000032:analysis*; D007700:Kinetics; D010446:Peptide Fragments / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42)