Application of magnetic chromatography to the isolation of lysosomes from fibroblasts of patients with lysosomal storage disorders.
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.
Diettrich
O
O
Institute of Child Health, London, UK. o.diettrich@ich.ucl.ac.uk
Mills
K
K
Johnson
A W
AW
Hasilik
A
A
Winchester
B G
BG
D002458:Cell Fractionation; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods*; D003911:Dextrans / Q000737:chemistry / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004705:Endocytosis; D005347:Fibroblasts; D006801:Humans; D007501:Iron / Q000737:chemistry; D016464:Lysosomal Storage Diseases / Q000473:pathology*; D008247:Lysosomes; D008280:Magnetics; D008928:Mitochondria / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*
D003911:Dextrans; D007501:Iron; D019158:N-Acetylneuraminic Acid