Covalent Immobilization of Unilamellar Liposomes in Gel Beads for Chromatography

✍ Scribed by Qing Yang; Xue-Ying Liu; Makoto Yoshimoto; Ryoichi Kuboi; Jun Miyake

• Publisher: Elsevier Science
• Year: 1999
• Tongue: English
• Weight: 94 KB
• Volume: 268
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1998.3070

✦ Synopsis

For immobilized (proteo)liposome chromatography, unilamellar liposomes were covalently bound within gel beads that had been activated by CNBr, N-hydroxysuccinimide, tresyl, or chloroformate. Liposomes composed of phosphatidylcholine (PC) and 2 mol% of amino-containing lipid (phosphatidylethanolamine-caproylamine) were immobilized in the activated gels at 5-35 mol lipid/ml gel and yields of 11-70%. The highest immobilized amount was found in chloroformate-activated TSK G6000PW gel, which contains large pore size (>100 nm). Liposomes composed of PC alone could also be attached to the chloroformate-activated gels at 33-42 mol/ml gel and yields of 58 -65%, probably by crosslinking of the phosphate moiety of phospholipid with the active group of the adsorbent. Liposomes prepared by various phospholipids with or without amino-containing lipids can generally be immobilized in the chloroformate-activated gels. The covalently bound liposomes were characterized by their high stability, unilamellarity, permeability of the membranes, and drug-membrane partition properties. A stable membrane phase was constructed for chromatographic experiments to be performed under extreme elution conditions.