Purification of alpha-amylase by specific elution from anti-peptide antibodies

Appl Microbiol Biotechnol. 1997 May;47(5):521-4. doi: 10.1007/s002530050966.

Abstract

Chimeric alpha-amylase, produced by recombinant yeast cells, was purified by immunoaffinity chromatography by use of an anti-peptide antibody and an eluent containing an antigen peptide. Chimeric alpha-amylase was adsorbed by the antibody against the peptide corresponding to the C-terminal region of target alpha-amylase, and specifically eluted by the eluent containing the antigen peptide used for immunization. A low concentration of the peptide could competitively elute adsorbed alpha-amylase, and the rate-limiting step of the elution was mass transfer of desorbed alpha-amylase. With this specific method, target proteins can be effectively eluted, and highly purified under mild conditions, from the antibody ligand showing a high-affinity for the adsorption step

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Animals
  • Antibodies / immunology
  • Rabbits
  • Recombinant Fusion Proteins / isolation & purification
  • alpha-Amylases / immunology
  • alpha-Amylases / isolation & purification*

Substances

  • Antibodies
  • Recombinant Fusion Proteins
  • alpha-Amylases