Reconstituting viral glycoproteins into large phospholipid vesicles

A method for the efficient reconstitution of a viral glycoprotein into large phospholipid vesicles with large internal aqueous spaces, which method comprises in combination:

• (A) extracting out the desired viral glycoprotein from viral particles with an extraction buffer comprising an alkyl glucoside;
• (B) mixing the extract with a phospholipid solution and deriving a cochleate intermediate in the form of a precipitate utilizing a direct calcium dialysis procedure;
• and
• (C) forming large phospholipid vesicles with integrated viral glycoprotein in a biologically active state by contacting the cochleate intermediate of (B) with an EDTA solution utilizing a rotary dialysis procedure against a buffered EDTA solution.

A method for the efficient reconstitution of membrane proteins into large phospholipid vesicles with large internal aqueous spaces, which method comprises in combination:

• (A) extracting out the desired membrane protein from a source biological material with an extraction buffer comprising a detergent;
• (B) mixing the extract with a negatively charged phospholipid, removing detergent and contacting with a calcium solution to form a cochleate intermediate, said cochleate intermediate being derived in the form of a precipitate utilizing a direct calcium dialysis procedure involving removing detergent simultaneously with said contacting with said calcium solution;
• and
• (C) forming large phospholipid vesicles with integrated membrane protein in a biologically active state by contacting a solution of the cochleate intermediate of (B) with a calcium chelating solution.