US 12,228,573 B2
Method of isolating and detecting exosomal biomarkers of glioblastoma malignancy
Abhimanyu Thakur, Kowloon (HK); Wing Kar Li, Kowloon (HK); and Youngjin Lee, Kowloon (HK)
Assigned to CITY UNIVERSITY OF HONG KONG, Kowloon (HK)
Filed by City University of Hong Kong, Kowloon (HK)
Filed on Jun. 1, 2021, as Appl. No. 17/335,605.
Prior Publication US 2022/0381782 A1, Dec. 1, 2022
Int. Cl. G01N 33/574 (2006.01)
CPC G01N 33/57407 (2013.01) [G01N 2333/70585 (2013.01); G01N 2333/70596 (2013.01)] 1 Claim
OG exemplary drawing
 
1. A method of identifying and treating a subject having glioblastoma comprising:
(a) identifying the subject by,
(i) obtaining a sample of blood or cerebrospinal fluid-derived extracellular vesicles (EVs) from the subject;
(ii) mixing the sample with a plurality of streptavidin-coated magnetic beads so as to separate Epidermal Growth Factor Receptor Variant-III (EGFRviii)-containing EVs from the sample, wherein, each streptavidin-coated magnetic bead is further linked with biotin, which is in turn bound to an anti-EGFRviii antibody;
separating the EGFRviii-containing EVs from the sample by,
(ii-1) using a magnet to move the streptavidin-coated magnetic beads bound EGFRviii-containing EVs away from the sample;
(ii-2) using a release buffer solution to break the bonding between the EGFRviii-containing EVs and the anti-EGFRviii antibody on the streptavidin-coated magnetic beads; and
(ii-3) using the magnet to remove the streptavidin-coated magnetic beads from the EGFRviii-containing EVs,
(iii) affixing the separated EGFRviii-containing EVs on a titanium nitride substrate having a layer of biotinylated Cluster of Differentiation (CD) 63 antibody coated thereon, in which the separated EGFRviii-containing EVs are affixed on the titanium nitride substrate via the attraction between CD63 of the separated EGFRviii-containing EVs and the biotinylated CD63 antibody; and
(iv) characterizing the expression of CD44 and CD133 on the separated EGFRviii-containing EVs via surface plasmonic resonance or atomic force microscopy; and
(b) administering a treatment to the subject when the expression of CD44 and CD133 are found on the separated EGFRviii-containing EVs in step (a), wherein the treatment is selected from the group consisting of chemotherapy, radiation, surgery, and a combination thereof.