Rapid Preparation of Giant Unilamellar Vesicles

We report here a rapid evaporation method that produces in high yield giant unilamellar vesicles up to 50 $\mu $m in diameter. The vesicles are obtained after only 2 min and can be prepared from different phospholipids, including L-$\alpha $-phosphatidylcholine (lecithin), dipalmitoleoyl L-$\alpha $-phosphatidylcholine, and $\beta $-arachidonyl $\gamma $-palmitoyl L-$\alpha $-phosphatidylcholine. Vesicles can be produced in distilled water and in Hepes, phosphate, and borate buffers in the pH range of 7.0 to 11.5 with ionic strengths up to 50 mM. The short preparation time allows encapsulation of labile molecular targets or enzymes with high catalytic activities. Cell-sized proteoliposomes have been prepared in which $\gamma $-glutamyltransferase (EC 2.3.2.2) was functionally incorporated into the membrane wall.