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Updated: May 20, 2026
A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation
Published on: March 27, 2017
[Optimization of expression and purification protocol for human scFv antibody against β-amyloid peptide]
Peng Xie1, Yu-xiao Wang, Rong-kai Gao
1Central Laboratory, Navy General Hospital, Beijing 100048, China. woodcarvingparlita@hotmail.com
Aim:
To optimize the expression and purification protocol for human scFv antibody against-amyloid peptide.
Methods:
Expression of E3 scFv was induced by different concentrations of IPTG under the fixed condition of time period and temperature, and the optimal concentration of IPTG was determined by SDS-PAGE analysis on E3 scFv expression level. Furthermore, elution buffer with different concentrations of imidazole was used for pre-eluting to determine the optimal pre-eluting condition by Western blotting against E3 scFv.
Results:
We obtained the highest expression of E3 scFv after 18 h induction with 0.1 mmol/L IPTG under 20 Degrees Celsius. In addition, Western blotting indicated the highest purity of E3 scFv when the resin was pre-eluted with the buffer containing 10 mmol/L imidazole.
Conclusion:
Through optimizing the above mentioned conditions, we established an improved strategy for high expression and efficient purification of E3 scFv, which paves the way for further research.
