US 11,911,713 B2
Exosome isolation method by two phase fluid system
Fikrettin Sahin, Istanbul (AR); Pakize Neslihan Tasli, Istanbul (AR); Oguz Kaan Kirbas, Istanbul (AR); and Batuhan Turhan Bozkurt, Istanbul (AR)
Assigned to YEDITEPE UNIVERSITESI, Istanbul (TR)
Appl. No. 16/969,997
Filed by YEDITEPE UNIVERSITESI, Istanbul (TR)
PCT Filed Feb. 15, 2019, PCT No. PCT/TR2019/050103
§ 371(c)(1), (2) Date Aug. 14, 2020,
PCT Pub. No. WO2019/160519, PCT Pub. Date Aug. 22, 2019.
Claims priority of application No. 2018/02164 (TR), filed on Feb. 15, 2018.
Prior Publication US 2021/0101093 A1, Apr. 8, 2021
Int. Cl. B01D 15/34 (2006.01); A61K 35/16 (2015.01); A61K 35/20 (2006.01); A61K 35/22 (2015.01); A61K 35/38 (2015.01); A61K 35/50 (2015.01); A61K 35/52 (2015.01)
CPC B01D 15/34 (2013.01) [A61K 35/16 (2013.01); A61K 35/20 (2013.01); A61K 35/22 (2013.01); A61K 35/38 (2013.01); A61K 35/50 (2013.01); A61K 35/52 (2013.01)] 21 Claims
OG exemplary drawing
 
1. An exosome isolation method by a two phase fluid extraction system for obtaining exosomes from cytoplasm inner and outer fluids, comprising the steps of
preparing a fluid sample containing the exosomes,
removing contaminants in the fluid sample,
mixing the fluid sample with an isolation solution containing PEG and dextran to form a mixture for isolation,
separating the mixture for isolation into two phases by using the two phase fluid extraction system,
discarding a supernatant upper phase of the two phases formed in the mixture for isolation, in order to remove proteins and undesired substances therein,
removing dextran from the lower phase containing the exomes, wherein removing dextran comprises:
adding a purification solution containing monohydric alcohol or acetone to remove dextran from the lower phase containing the exosomes,
removing the dextran from the lower phase containing the exosomes via a filtration,
removing the dextran precipitated by the purification solution,
removing the purification solution from the exosomes and thereby obtaining exosomes in a pure form in a collected phase from the mixture for isolation;
mixing the collected phase from the mixture for isolation with a washing solution to form a mixture for washing,
separating the mixture for washing into two phases using the two phase fluid extraction system,
discarding a supernatant of the two phases formed in the mixture for washing and collecting a lower phase containing the exosomes.