Purification of recombinantly expressed and cytotoxic human amyloid-beta peptide 1–42

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

“…Here, we used our rAb to confirm the Cu 2+ interaction. In comparison, most cross peaks in our spectrum were consistent with that in Hou et al work except for the obvious missing side chain peak of R5 and some differences in the ones situated between 7.8 and 8.0 ppm in 1 H and 120-125 ppm in 15 N. Upon addition of one molar equivalent of Cu 2+ , line broadening caused disappearance of several peaks corresponding to the backbone NH signals of residues E3-V18 and the side chain of Q15 in the spectra of Ab (Fig. 8A).…”

“…Currently, there are several protocols detailing production of recombinant Ab [12][13][14][15][16]. Several of these protocols were developed to produce full-length Ab with the yield of 1-L bacterial culture ranging from 4 to 24 mg [12,[14][15][16][17][18][19][20][21].…”

“…Several of these protocols were developed to produce full-length Ab with the yield of 1-L bacterial culture ranging from 4 to 24 mg [12,[14][15][16][17][18][19][20][21]. Most constructs in these studies include a soluble protein as a fusion tag to prevent Ab aggregation and cytotoxicity to the cells.…”

Interdisciplinary challenges and promising theranostic effects of nanoscience in Alzheimer's disease

“…Ab is produced either by solid phase synthesis or by recombinant techniques. 40 The preparations may be contaminated with salts or other factors that affect the dissolution. Fibrillogenesis is affected by the preparation conditions such as pH, temperature, solvent, concentration, etc.…”

Alzheimer´s Disease-associated Aβ42 Peptide: Expression and Purification for NMR Structural Studies

“…Modifications included oxidation on the methionine 35 side chain of Aβ42 to methionine sulfoxide (Met-35(ox)Aβ42) [6,13], Aβ42 fused to a tag [14][15][16], and Aβ42 containing unnatural mutations in its sequence [6,17] or additional N-terminal residues [18][19][20][21]. More recently, several strategies have led to the production of wild-type Aβ42 with satisfactory yields [22][23][24][25][26][27][28], as well as isotopically labeled 15 N and 15 N, 13 C Aβ42 [29,30].…”