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

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

    研究成果: Article

    76 引用 (Scopus)

    抄録

    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.

    元の言語English
    ページ(範囲)1547-1552
    ページ数6
    ジャーナルBiotechnology Progress
    19
    発行部数5
    DOI
    出版物ステータスPublished - 2003 9

    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

    これを引用

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

    :: Biotechnology Progress, 巻 19, 番号 5, 09.2003, p. 1547-1552.

    研究成果: Article

    Sou, Keitaro ; Naito, Yoshiyasu ; Endo, Taro ; Takeoka, Shinji ; Tsuchida, Eishun. / Effective Encapsulation of Proteins into Size-Controlled Phospholipid Vesicles Using Freeze-Thawing and Extrusion. :: Biotechnology Progress. 2003 ; 巻 19, 番号 5. pp. 1547-1552.
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