US 12,282,022 B2
Nanoplasmonic quantification of tumor-derived extracellular vesicles in plasma microsamples for detection and treatment monitoring
Ye Hu, Scottsdale, AZ (US); Jia Fan, Pearland, TX (US); Kai Liang, Beijing (CN); Fei Liu, Cupertino, CA (US); and Dali Sun, Tempe, AZ (US)
Assigned to THE METHODIST HOSPITAL, Houston, TX (US)
Appl. No. 16/469,082
Filed by THE METHODIST HOSPITAL, Houston, TX (US)
PCT Filed Dec. 28, 2017, PCT No. PCT/US2017/068768
§ 371(c)(1), (2) Date Jun. 12, 2019,
PCT Pub. No. WO2018/126043, PCT Pub. Date Jul. 5, 2018.
Claims priority of provisional application 62/440,494, filed on Dec. 30, 2016.
Prior Publication US 2020/0096516 A1, Mar. 26, 2020
Int. Cl. G01N 33/574 (2006.01); G01N 33/543 (2006.01)
CPC G01N 33/57438 (2013.01) [G01N 33/54346 (2013.01)] 8 Claims
 
1. A method of detecting a cancer specific protein that is enriched on an extracellular vesicle (EV), comprising:
contacting a sample comprising the EV with a surface conjugated with a first antibody directed to a first molecule on the EV to capture the EV on the surface;
removing the remainder of the sample from the surface to generate separated EV captured on the surface;
incubating the separated EV captured on the surface with a first set of nanoparticles conjugated with a second antibody directed to the cancer specific protein on the EV and incubating the separated EV captured on the surface with a second set of nanoparticles conjugated with a third antibody directed to a second molecule on the EV, wherein the first molecule and the second molecule are selected from the group consisting of CD63, CD81, CD9, CD24, CD26, CD10, tumor susceptibility gene 101 (TSG101), heat shock 70 kDa protein 4 (HSP70), Rab-5b, programmed cell death 6-interacting protein (AIP1/Alix), aquaporin 2 (AQP2), vascular endothelial growth factor receptor 1 (FLT1), N-glycan containing a fucose residue, phosphocholine, phosphatidylserine, and sphingomyelin; and
detecting whether said cancer specific protein is present in the sample by detecting binding between the second antibody and the cancer specific protein on the EV, wherein dual binding of the two antibody-conjugated nanoparticles to the EV produces nanoplasmons and said nanoplasmons are detected,
wherein the cancer specific protein is ephrin type-A receptor 2 (EphA2), and wherein the EphA2 is associated with pancreatic cancer.