A novel method for expression and purification of authentic amyloid-β with and without (15)N labels

Amyloid-β (Aβ) is a major constituent in the senile plaques of patients with Alzheimer's disease (AD). Aβ has been intensively studied in amyloid research; however, challenges posed by data reproducibility arise from purity of synthetic Aβ and high expense for its isotope-labeling. The difficulties motivate development and optimization of recombinant Aβ (rAβ) production. Here, we report a new procedure to express and purify high quality rAβ40 from Escherichia coli. The new Aβ construct expressed insoluble Aβ fused with an N-terminal histidine-tag connected by a linker harboring TEV protease cut site. After purification and partial refolding, the fusion tag was removed by TEV protease cleavage, immobilized metal affinity chromatography (IMAC), and reversed phase-HPLC purification with a yield of 3.5 mg/L culture with and without (15)N label. The rAβ adopts classic amyloid fibrillization and is capable of binding to its clinical relevant metal ions.