Large scale isolation of human erythrocyte membranes by high volume molecular filtration

Terrone L. Rosenberry, Jeffrey F. Chen, Mary M.L. Lee, Thomas A. Moulton, Philip Onigman

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

A molecular filtration procedure for preparing large quantities of human erythrocyte ghost membranes is presented. Hemolysate ghost membranes are rapidly cycled in the retantate channel of the filtration apparatus, while hemoglobin is removed s it pass through Pellicon filters into the filtrate. Several-liter quantities of washed packed erythrocytes can be processed in a few hours with this system and the filtration procedure does not appear to alter erythrocyte or ghost membranes. Intact erythrocytes in isotonic solution can be circulated through the retentate channel for 16 h with only 3% hemolysis and with preservation of their orginal morphology in scanning electron microscopy. Ghost membranes isolated by the procedure are virtually identical in morphology, polypeptide composition and acetylcholinesterase content to membranes isolated by conventional centrifugation techniques.

Original languageEnglish (US)
Pages (from-to)39-48
Number of pages10
JournalJournal of Biochemical and Biophysical Methods
Volume4
Issue number1
DOIs
StatePublished - Jan 1981

Keywords

  • acetylcolinesterase
  • erythrocyte membranes
  • hemolysis
  • membrane filtration

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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