Granted Patent
B2
US 12,429,486 · App. 18/060,455 · Granted Sep 30, 2025
Use of Tenascin-C as an extracellular marker of tumor-derived microparticles
Assignee:
NX PHARMAGEN INC.
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Abstract
The present disclosure provides methods for isolating tumor-derived microparticles from a subject for analysis, specifically by isolating Tenascin-C positive microparticles from a sample from the subject to obtain tumor-derived microparticles. Methods for determining the expression status of biomarkers in the tumor-derived microparticles and methods for determining additional characteristics of the tumor-derived microparticles are also provided.
Claims (14)
1. A method of isolating brain tumor-derived microparticles from a human subject for analysis, the method comprising:
(1) providing a sample comprising bodily fluid from the human subject;
(2) subjecting the sample to treatment with a LaurA solution, wherein the solution comprises between 5% and 10% LaurA;
(3) isolating bodily fluid-derived microparticles from the sample, wherein the bodily-fluid derived microparticles comprise endosome-derived exosomes, plasma membrane-derived shedding vesicles, apoptotic bodies, outer membrane vesicles (OMVs), or a combination thereof, released from cells in the human subject, and wherein the isolating of the bodily fluid-derived microparticles comprises:
(a) subjecting the sample to ultracentrifugation or size exclusion chromatography (SEC); or
(b) performing an affinity pulldown with the sample using a Vn96 peptide or a Heladonin peptide as an affinity reagent;
(4) isolating antibody-purified microparticles from the bodily fluid-derived microparticles using an anti-Tenascin-C antibody; and
(5) analyzing the antibody-purified microparticles to determine the expression status of at least two biomarkers selected from EGFRvIII, 14-3-3, PPIX, and glycoporin A, wherein the at least two biomarkers selected from a presence of EGFRvIII, a presence of 14-3-3, a presence of PPIX, and an absence of glycoporin A are indicative of the antibody-purified microparticles being brain-tumor derived microparticles.
2. The method of claim 1 , wherein the SEC is performed with a solid support and a mobile phase added to the solid support.
3. The method of claim 2 , wherein the solid support comprises a column.
4. The method of claim 2 , wherein the mobile phase is water.
5. The method of claim 1 , wherein the expression status of the at least two biomarkers are determined by an immune-based assay or mass spectrometry.
6. The method of claim 5 , wherein the immune-based assay is an immunosorbent assay (ELISA), a radioimmune assay (RIA), a Western blot assay, or a slot blot assay.
7. The method of claim 1 , wherein the anti-Tenascin-C antibody is 81C6.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 26, 2024
From: EZRIN, ALAN M.; GRIFFITHS, STEVEN G.
To: NX PHARMAGEN INC.
Reel/Frame 067243/0252 →
Continuity (3)
Continuation
14893042
Provisional Application
61825951
· May 21, 2013
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