Tissue-derived extracellular vesicles and their use as diagnostics
AssigneeExocure Sweden AB
InventorsJan Lötvall, Su Chul Jang, Rossella Crescitelli
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Abstract
The present disclosure relates to a method of isolating extracellular vesicles directly from human tissues. The invention further relates to a method of identifying disease and tissue specific membrane proteins on extracellular vesicles by membrane isolation and proteomic analysis. The invention further relates to methods of diagnosing diseases by capturing extracellular vesicles by the use of disease specific membrane proteins from body fluids, and detecting or analyzing molecular signatures (proteome, DNA, and RNA) on captured extracellular vesicles. Moreover, the present invention relates to kits, apparatus and software required for implementing aforementioned methods.
Claims11
A method comprising the steps of:
- (a) fragmenting a human tissue into a plurality of pieces;
- (b) incubating the plurality of pieces with one or more enzymes, wherein the one or more enzymes are selected from a group of proteases including a matrix metalloproteinase, collagenases, and papain and nucleases including DNase, RNase, and Benzonase, wherein the incubating releases extracellular vesicles from the plurality of pieces, thereby generating a mixture of the extracellular vesicles, the plurality of pieces, and the one or more enzymes;
- (c) isolating the extracellular vesicles from the mixture;
- (d) treating the extracellular vesicles with an aqueous solution, wherein the aqueous solution has a pH in a range of 9 to 14 to generate membranes;
- (e) isolating the membranes;
- and
- (f) identifying proteins on the membranes by employing mass spectrometry to identify tissue specific membrane proteins.
Dependencies are inferred from the claim text.