反応 #568462

反応方程式

chloroform isopropyl ether methanol

DPPC

cholesterol

DPPG

Alachlor DPPE

→

Alachlor

溶媒

反応条件

後処理

1. 1
   その他Liposomes were formed by the reversed-phase evaporation method
2. 2
   その他This mixture was sonicated for 5 minutes under a low flow of nitrogen
3. 3
   その他The organic phase was removed under vacuum on a rotary evaporator at 40° until all frothing
4. 4
   workup.ADDITIONAn additional 1.3 ml aliquot of the dye solution was added
5. 5
   その他except that the usual sonication step
6. 6
   その他Finally, to remove any unencapsulated dye
7. 7
   ろ過filtered on a 1×14 cm Sephadex G-50 column
8. 8
   その他dialyzed overnight against TBS at 4° C
9. 9
   その他stored at 4° C.
10. 10
    その他of 9 months

実験手順

Liposomes were formed by the reversed-phase evaporation method, as described in Szoka, et al., Biochim. Biophys. Acta, 601 (1980) 559, and O'Connell, et al., Anal. Chem., 31 (1985) 142, the disclosures of which are hereby incorporated by reference, from a mixture of DPPC, cholesterol, DPPG, and Alachlor-DPPE conjugate in a molar ration of 5:5:0.5:0.01. Forty-three μmol of this mixture were dissolved in 4.2 ml of a solvent mixture containing chloroform-isopropyl ether-methanol (6:6:1,v/v). This solution was warmed to 45° C. and 0.7 ml of the dye solution was added with swirling. This mixture was sonicated for 5 minutes under a low flow of nitrogen. The organic phase was removed under vacuum on a rotary evaporator at 40° until all frothing had stopped. An additional 1.3 ml aliquot of the dye solution was added, and the liposomes were then sequentially extruded twice through each of two polycarbonate filters of decreasing pore sizes of 1.0 μm and 0.4 μm. The diameters of the liposome preparations were measured by laser scattering in a LA-900 particle size distribution analyzer (Horiba, Irvine, Calif.), using the manufacturers method, except that the usual sonication step was omitted to avoid lysis (rupture) of the liposomes. Finally, to remove any unencapsulated dye, the liposomes were gel filtered on a 1×14 cm Sephadex G-50 column and dialyzed overnight against TBS at 4° C. When stored at 4° C., there was no significant leakage of dye over a period of 9 months as described below.

出典