Combined use of micro-preparative gel electrophoresis and reversed-phase high-performance liquid chromatography for purification of amyloid beta peptides deposited in brains of Alzheimer's disease patients

J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Apr 5;769(2):363-70. doi: 10.1016/s1570-0232(02)00024-7.

Abstract

A new micro-technique is developed for purification of amyloid beta peptides (A beta) extracted from brain tissues of patients with Alzheimer's disease (AD). It includes SDS-polyacrylamide gel electrophoresis of the extracted brain tissue material, electroblotting onto supporting membranes, and reversed-phase HPLC of the proteins eluted from membranes. By this technique, the extracted A beta are first separated electrophoretically from the higher and lower molecular mass tissue components, and then purified by reversed-phase HPLC from the contaminants having similar molecular masses, but different retention times on the column. In contrast to the common large-scale isolation procedures employing density gradient centrifugation, enzymatic digestions and size-exclusion chromatography, the developed micro-technique might be applied for biochemical analysis of A beta contained in small AD brain tissue specimens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / metabolism*
  • Amyloid beta-Peptides / isolation & purification*
  • Amyloid beta-Peptides / metabolism
  • Blotting, Western
  • Brain / metabolism*
  • Chromatography, High Pressure Liquid / methods*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Humans

Substances

  • Amyloid beta-Peptides