Hi, I am planning to synthesize DOPC and or DOTAP (mixed and pure) vesicles using sonication. Any suggestions on cleaning the glassware before and after synthesis will be really helpful as I did not find a single paper mentioning those steps. Also, any tips and tricks or ratio for synthesis will be highly appreciated.
Thanks in advance.
You don't need to plasma clean or acid clean your glassware for this purpose. Most likely, clean with ethanol or acetone water solution. But, make sure you dry the glassware after using those solutions. Tips regarding the synthesis would be - make sure to use rotary evaporator to obtain a good thinfilm of lipids before pouring the appropriate buffer to get liposomes or vesicles. Masking the charges of the lipid DOTAP might be useful when you try to produce 100% DOTAP vesicles. I hope this information is helpful.
Hi Dhiraj Kumar just to add to Naresh's answer is that after cleaning the glassware as he described, you can make liposomes by drying a lipid solution in chloroform in a glass vial, add buffer and then use a simple sonicating waterbath to make the liposomes.
Hi Dhiraj, just wondering if you had success with your liposome formulation and if you have recommendations for storing DOTAP:DOPE-Chloroform solution e.g. type of glassware and environment? thanks
I have been trying to synthesize DOTAP ( cationic ) and DOPE ( neutral) based liposomes using (50% of DOTAP in final lipid volume in chloroform ). After thin film formation and hydrating the film using PBS or HEPES buffer ( tried both ) , sonication ( 15 min at 40 deg )of the resulting hydrated lipid mixture is forming aggregates which cannot be dispersed any further. Have you guys tried to make them or have any tips for better formation of liposomes
Stephen John Mccleary, i store lipid solutions in chloroform-methanol solution at -20c.
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