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Permeability Study of Phospholipid Liposomes Prepared by Reverse Phase Evaporation Method
임근풍,최명언,Lim, Keun-Poong,Choi, Myung-Un 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.6
역상 증발 방법으로 제조된 인지질 리포좀의 특성 연구를 위하여 리포좀의 투과성을 6-carboxyfluorescein(6-CF)의 유출로써 관찰하였다. 젤 여과 후에 시간에 따른 흡광도의 변화를 492 nm에서 측정하였다. 투과성의 변수로서 포획된 6-CF의 절반 양이 유출되는 반감기를 택했다. 이온세기가 150 mM NaCl이고 pH가 6.5인 10mM MES 완충용액에서 27 nmol/ml의 PC liposome의 반감기는 $37^{\circ}C$에서 9분이었다. 투과성에 영향을 미칠 수 있는 여러 가지 요인들 즉 pH, 온도, 이온세기 및 금속이온 등을 조사하였다. PC liposome으로부터 얻어진 이러한 결과들을 서로 다른 인지질로 만든 혼성 리포좀에도 확장시켰다. Head group을 변화시킴으로써 두 가지 성분으로 이루어진 생체막 계의 안정성이 확인되었으며, 그의 혼성 리포좀의 투과성에 대한 $Ca^{2+}$의 선택적인 영향도 검토되었다. In order to characterize phospholipid liposomes prepared by reverse-phase evaporation method, the permeability of liposomes was monitored by the efflux of 6-carboxyfluorescein (6-CF). The changes of optical density with time were measured spectrophotometrically at 492 nm after gel filtration. From the simple hyperbolic curve obtained, the half-time of efflux of 6-CF was estimated as the paramter of the permeability. Half-time of efflux of phosphatidylcholine (PC) liposome (27 nmol/ml) suspended in 10 mM MES (pH 6.5) buffer with the ionic strength of 150 mM NaCl at $37^{\circ}C$ was 9 min. Various factors which could affect the permeability were examined including pH, temperature, ionic strength, and metal ions. These results from the PC liposome were extended to the mixed liposomes with different phospholipid head groups. The head group variation confirmed the stability of the binary membrane system. Furthermore, the selective effect of $Ca^{2+}$ ion on the permeability of mixed liposomes was examined.
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역상 증발 방법으로 제조된 인지질 리포좀의 투과성 연구
임근풍,최명언 ( Keun Poong Lim,Myung Un Choi ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.6
In order to characterize phospholipid liposomes prepared by reverse-phase evaporation method, the permeability of liposomes was monitored by the efflux of 6-carboxyfluorescein (6-CF). The changes of optical density with time were measured spectrophotometrically at 492 nm after gel filtration. From the simple hyperbolic curve obtained, the half-time of efflux of 6-CF was estimated as the paramter of the permeability. Half-time of efflux of phosphatidylcholine (PC) liposome (27 nmol/ml) suspended in 10 mM MES (pH 6.5) buffer with the ionic strength of 150 mM NaCl at 37℃ was 9 min. Various factors which could affect the permeability were examined including pH, temperature, ionic strength, and metal ions. These results from the PC liposome were extended to the mixed liposomes with different phospholipid head groups. The head group variation confirmed the stability of the binary membrane system. Furthermore, the selective effect of Ca^(2+) ion on the permeability of mixed liposomes was examined.
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약물운반체 개발을 위한 각종 인지질 베시클들의 비교 제조 및 특성실험
이충희,임근풍,박홍식,정관영,최명언 ( Choong Hee Lee,Keun Poong Lim,H . S . Park,Kwan Young Jung,Myung Un Choi ) 생화학분자생물학회 1986 BMB Reports Vol.19 No.4
In order to develop an efficient drug-delivery system, various phospholipid vesicles were prepared and their basic properties were compared. Phospholipids used for vesicle preparation were extracted from rat brain by Folch method and separated by DEAE-cellulose and silicic acid column. Some of phospholipids were also synthesized from egg phosphatidylcholine by means of the phospholipase D. Small unilamellar vesicle, large unilamellar vesicle, and plurilamellar vesicle were prepared by the methods of sonication, reverse-phase evaporation, and ether emulsification respectively 6-Carboxyfluorescein techniques were used for estimation of the encapsulation capacity and permeability of the prepared vesicles.
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약물운반체 개발을 위한 각종 인지질 베시클들의 비교 제조 및 특성실험
이충희,임근풍,박흥식,정관영,최명언,Lee, Choong-Hee,Lim, Keun-Poong,Park, H.S.,Jung, Kwan-Young,Choi, Myung-Un 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.4
약물운반체 개발을 위한 기초 실험으로 여러 종류의 인지질 베시클들을 제조하여 그 특성 들을 비교 연구하였다. 사용된 인지질들은 쥐 뇌로부터 Folch 방법으로 추출하고 DEAE cellulose와 silicic acid column으로 분리하였으며, 경우에 따라선 phospholipase D를 이용하여 달걀 phosphatidylcholine으로부터 합성하였다. Small unilamellar vesicle은 sonication 방법으로, large unilamellar vesicle은 reverse-phase evaporation 방법으로, plurilamellar vesicle은 소량의 에텔로 emulsify한 후 수용액을 넣어서 각각 제조하였다. 제조된 베시클들의 포획 능력과 투과성은 6-carboxyfluorescein을 이용하여 추정하였다. In order to develop an efficient drug-delivery system, various phospholipid vesicles were prepared and their basic properties were compared. Phospholipids used for vesicle preparation were extracted from rat brain by Folch method and separated by DEAE-cellulose and silicic acid column. Some of phospholipids were also synthesized from egg phosphatidylcholine by means of the phospholipase D. Small unilamellar vesicle, large unilamellar vesicle, and plurilamellar vesicle were prepared by the methods of sonication, reverse-phase evaporation, and ether emulsification respectively. 6-Carboxyfluorescein techniques were used for estimation of the encapsulation capacity and permeability of the prepared vesicles.
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