title: [Optimization of expression and purification protocol for human scFv antibody against  β-amyloid peptide].
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors: Central Laboratory, Navy General Hospital, Beijing 100048, China. woodcarvingparlita@hotmail.com
authors: Peng
authors: P
authors: Xie
authors: Yu-xiao
authors: YX
authors: Wang
authors: Rong-kai
authors: RK
authors: Gao
authors: Li-jun
authors: LJ
authors: Zhou
mesh_terms: D016229:Amyloid beta-Peptides / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D016229:Amyloid beta-Peptides; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies