Granted Patent B2

US 12,282,022 · App. 16/469,082 · Granted Apr 22, 2025

Inventors: Ye Hu (Scottsdale, AZ); Jia Fan (Pearland, TX); Kai Liang (Beijing, CN); Fei Liu (Cupertino, CA); Dali Sun (Tempe, AZ)

Assignee: THE METHODIST HOSPITAL

G01N33/57438G01N33/54346

Abstract

Claims (13)

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.

2. The method of claim 1 , wherein the nanoparticles comprise a plurality of nanospheres, a plurality of nanorods, or a combination thereof.

3. The method of claim 2 , wherein the detecting step comprises detecting binding between the second antibody and the EV and detecting binding between the third antibody and the EV.

4. The method of claim 2 , wherein the nanoparticles comprise gold.

5. The method of claim 2 , wherein the first antibody is different from the third antibody.

6. The method of claim 2 , wherein the first antibody is the same as the third antibody.

7. The method of claim 2 , wherein the second antibody is different from the third antibody.

8. The method of claim 2 , wherein the first nanoparticle is different from the second nanoparticle.

Assignments (1)

Continuity (2)

References Cited (75)