US 12,109,309 B2
Internally fixed lipid vesicle
Che-Ming Jack Hu, Taipei (TW); Hui-Wen Chen, Taipei (TW); Yuan-I Chen, Taipei (TW); Chen-Ying Chien, Taichung (TW); and Jung-Chen Lin, New Taipei (TW)
Assigned to Academia Sinica, Taipei (TW)
Appl. No. 16/322,014
Filed by Academia Sinica, Taipei (TW)
PCT Filed Jul. 28, 2017, PCT No. PCT/US2017/044326
§ 371(c)(1), (2) Date Jan. 30, 2019,
PCT Pub. No. WO2018/026644, PCT Pub. Date Feb. 8, 2018.
Claims priority of provisional application 62/369,440, filed on Aug. 1, 2016.
Prior Publication US 2019/0170745 A1, Jun. 6, 2019
Int. Cl. A61K 9/127 (2006.01); C12N 5/078 (2010.01); G01N 33/50 (2006.01); G01N 33/543 (2006.01); G01N 33/556 (2006.01)
CPC A61K 9/127 (2013.01) [A61K 9/1275 (2013.01); A61K 9/1277 (2013.01); C12N 5/0634 (2013.01); G01N 33/5008 (2013.01); G01N 33/5432 (2013.01); G01N 33/556 (2013.01); G01N 2500/02 (2013.01); G01N 2500/10 (2013.01)] 20 Claims
 
1. A method of generating an internally fixed lipid vesicle, comprising:
providing a precursor lipid vesicle that contains an aqueous interior enclosed by a precursor lipid membrane, wherein the precursor lipid membrane of the precursor lipid vesicle is non-permeable to a crosslinker;
permeabilizing the precursor lipid membrane transiently to generate a permeable vesicle;
contacting the permeable vesicle with an inactive activatable crosslinker, whereby the inactive activatable crosslinker enters the permeable vesicle, wherein the inactive activatable crosslinker is a monomer or polymer;
allowing the permeable vesicle to return to a non-permeable vesicle;
removing any extravesicular crosslinker; and
activating the inactive activatable crosslinker to allow crosslinking to occur inside the non-permeable vesicle, whereby an internally fixed lipid vesicle containing a gelated interior is generated, and wherein the internally fixed lipid vesicle has a lipid membrane that is substantially identical to the precursor lipid membrane in one or more of membrane fluidity, membrane protein mobility, sensitivity to a surfactant, membrane permeability, membrane content, surface charge, and biological function.