[Optimization of expression and purification protocol for human scFv antibody against β-amyloid peptide].

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.