US 11,714,068 B2
Exosome production method
Hiroshi Shiku, Tsu (JP); Naohiro Seo, Tsu (JP); Tsuguhiro Kaneda, Tsu (JP); Junko Nakamura, Tsu (JP); and Kazunari Akiyoshi, Kyoto (JP)
Assigned to Mie University, Mie (JP)
Appl. No. 17/263,080
Filed by Mie University, Tsu (JP)
PCT Filed Jul. 31, 2019, PCT No. PCT/JP2019/029972
§ 371(c)(1), (2) Date Jan. 25, 2021,
PCT Pub. No. WO2020/027185, PCT Pub. Date Feb. 6, 2020.
Claims priority of application No. 2018-143200 (JP), filed on Jul. 31, 2018.
Prior Publication US 2021/0239664 A1, Aug. 5, 2021
Int. Cl. G01N 30/88 (2006.01); G01N 1/40 (2006.01)
CPC G01N 30/88 (2013.01) [G01N 1/405 (2013.01); G01N 1/4077 (2013.01); G01N 2001/4088 (2013.01)] 6 Claims
 
1. A method for preparing exosomes from a biological sample, the method comprising: (i) a step for performing centrifugal separation on a biological sample to remove contaminants from the sample, and then obtaining a supernatant; (ii) a step for subjecting the supernatant obtained in the step (i) to ultrafiltration by using a tangential flow filter having a nominal molecular weight cut-off (MWCO) of about 750 kDa and a pore size of 0.22 μm, and then concentrating the supernatant; and (iii) a step for subjecting a concentrated liquid obtained in the step (ii) to anion exchange column chromatography, wherein an anion exchanger used in the anion exchange column chromatography is a weakly basic anion exchanger, which retains selectivity based on both hydrophobic interaction and hydrophobic binding formation, and has a phenyl group, an amide bond, and a carboxyl group.