Application of magnetic chromatography to the isolation of lysosomes from fibroblasts of patients with lysosomal storage disorders

FEBS Lett. 1998 Dec 28;441(3):369-72. doi: 10.1016/s0014-5793(98)01578-6.

Abstract

A method for the purification of lysosomes from fibroblasts has been developed which uses endocytosis of superparamagnetic colloidal iron dextran particles followed by separation of the iron-containing lysosomes in a magnetic field. This permitted isolation of lysosomes from fibroblasts from patients with infantile sialic acid storage disorder and other lysosomal storage diseases in which a shift in lysosomal density induced by the storage material prevents purification by centrifugation in a Percoll gradient. The magnetic lysosomes isolated from these cells are very similar to those from normal cells as judged by lysosomal marker enzyme activity and 2D-PAGE analysis of the enriched proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Fractionation
  • Cells, Cultured
  • Chromatography, Affinity / methods*
  • Dextrans / chemistry
  • Dextrans / metabolism
  • Electrophoresis, Gel, Two-Dimensional
  • Endocytosis
  • Fibroblasts
  • Humans
  • Iron / chemistry
  • Lysosomal Storage Diseases / pathology*
  • Lysosomes*
  • Magnetics
  • Mitochondria / metabolism
  • N-Acetylneuraminic Acid / metabolism*

Substances

  • Dextrans
  • Iron
  • N-Acetylneuraminic Acid