Effective Encapsulation of Proteins into Size-Controlled Phospholipid Vesicles Using Freeze-Thawing and Extrusion

Keitaro Sou, Yoshiyasu Naito, Taro Endo, Shinji Takeoka, Eishun Tsuchida

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

We are aiming to improve the encapsulation efficiency of proteins in a size-regulated phospholipid vesicle using an extrusion method. Mixed lipids (1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), cholesterol, 1,5-dipalmitoyl-L-glutamate-N-succinic acid (DPEA), and 1,2-distearoyl-n-glycero-3-phosphoethanolamine-N-[monomethoxy poly(ethylene glycol) (5,000)] (PEG-DSPE) at a molar ratio of 5, 5, 1, and 0.033 were hydrated with a NaOH solution (7.6 mM) to obtain a polydispersed multilamellar vesicle dispersion (50 nm to 30 μm diameter). The polydispersed vesicles were converted to smaller vesicles having an average diameter of ca. 500 nm with a relatively narrow size distribution by freeze-thawing at a lipid concentration of 2 g dL-1 and cooling rate of -140°C min -1. The lyophilized powder of the freeze-thawed vesicles was rehydrated into a concentrated protein solution (carbonyl hemoglobin solution, 40 g dL-1) and retained the size and size distribution of the original vesicles. The resulting vesicle dispersion smoothly permeated through the membrane filters during extrusion. The average permeation rate of the freeze-thawed vesicles was ca. 30 times faster than that of simple hydrated vesicles. During the extrusion process, proteins were encapsulated into the reconstructed vesicles with a diameter of 250 ± 20 nm.

Original languageEnglish
Pages (from-to)1547-1552
Number of pages6
JournalBiotechnology Progress
Volume19
Issue number5
DOIs
Publication statusPublished - 2003 Sep 1

ASJC Scopus subject areas

  • Biotechnology

Fingerprint Dive into the research topics of 'Effective Encapsulation of Proteins into Size-Controlled Phospholipid Vesicles Using Freeze-Thawing and Extrusion'. Together they form a unique fingerprint.

  • Cite this