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

    Fingerprint

    encapsulation
    extrusion
    thawing
    Phospholipids
    phospholipids
    Carboxyhemoglobin
    Lipids
    Proteins
    Ethylene Glycol
    proteins
    Succinic Acid
    succinic acid
    lipids
    polyethylene glycol
    Phosphatidylcholines
    phosphatidylcholines
    glutamates
    Powders
    Glutamic Acid
    hemoglobin

    ASJC Scopus subject areas

    • Food Science
    • Biotechnology
    • Microbiology

    Cite this

    Effective Encapsulation of Proteins into Size-Controlled Phospholipid Vesicles Using Freeze-Thawing and Extrusion. / Sou, Keitaro; Naito, Yoshiyasu; Endo, Taro; Takeoka, Shinji; Tsuchida, Eishun.

    In: Biotechnology Progress, Vol. 19, No. 5, 09.2003, p. 1547-1552.

    Research output: Contribution to journalArticle

    Sou, Keitaro ; Naito, Yoshiyasu ; Endo, Taro ; Takeoka, Shinji ; Tsuchida, Eishun. / Effective Encapsulation of Proteins into Size-Controlled Phospholipid Vesicles Using Freeze-Thawing and Extrusion. In: Biotechnology Progress. 2003 ; Vol. 19, No. 5. pp. 1547-1552.
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