Materials, methods, and sampling devices for preserving and extracting extracellular vesicles in biofluids

A method for isolating extracellular vesicles present in whole blood, comprising:

• inputting a whole blood sample including one or more principal components and extracellular vesicles present therein into a biofluid card assembly;
• receiving the whole blood sample via a top lid of the biofluid card assembly with at least one opening formed therein,
• separating the extracellular vesicles via a biofluid separation membrane assembly disposed below and corresponding to the at least one opening and configured to separate the one or more principal components of the whole blood from remaining components of the whole blood including extracellular vesicles;
• and
• capturing the extracellular vesicle via at least one extracellular vesicle capture membrane having at least one extracellular vesicle capture agent formed thereon, each disposed below or embedded in the at least one extracellular vesicle capture membrane and corresponding to the at least one opening and adjacent the biofluid separation membrane assembly on one side and adjacent a bottom lid on a second side, wherein each of the at least one extracellular vesicle capture agent includes binding agent configured to bind to the extracellular vesicle having predetermined binding sites.