Antiapoe

The interaction between Aβ fibrils and ApoE was monitored using a BIAcore 3000 biosensor (GE Healthcare, Uppsala, Sweden) equipped with a CM5 sensor chip (GE Healthcare). Prior to fibril immobilization the dextran matrix on the sensor chip surface was activated with a mixture of 1-ethyl-3- (3-dimethylaminopropyl) carbodiamide (EDC) and N-hydroxysuccinimide (NHS). Aβ fibrils were immobilized at a density of 2000 response unit (RU) using standard amino coupling reagents and then deactivated according to the manufacture instruction. All SPR experiments were performed in degassed PBS at 25 °C. Fibrils were immobilized at a flow rate of 5 μL/min, whereas Aβ_1–40 monomeric peptide solution, ApoE4, Anti-ApoE (rabbit polyclonal, Cat# PA5–27088, ThermoFisher Scientific) and 15 nm protein-A gold beads (CMC Utrecht, Netherlands) were injected at a flow rate of 20 μL/min. All the steps were followed by a 5 min flow of buffer, and after the final step additionally washed by degassed distilled water for 5 min at a flow rate of 50 μL/min. Biosensor data were processed by BioLogic Software Scrubber2 and sensograms were made using GraphPad Prism 5.01.