Methods for isolation of biomarkers from vesicles
AssigneesHitachi Chemical Company, Ltd., Hitachi Chemical Company America, Ltd.
InventorMasato Mitsuhashi
Life of this patent
Estimating the term…
Abstract
Some embodiments provide for a device that collects vesicles and vesicle-like materials from biological fluids. Such devices include at least one sample loading region; at least one corresponding vesicle-capture material, wherein said vesicle-capture material includes glass-like materials; and at least one corresponding sample receiving region, wherein passage of the biological fluid from the sample loading region through the vesicle-capture material and into the sample receiving region results in the capture of vesicles. Additional embodiments provide for a method of isolating vesicles and vesicle-like materials from biological fluids.
Claims17
A method of isolating a biomarker from biological fluid, comprising:
- (a) obtaining a biological fluid sample comprising vesicles comprising at least one biomarker;
- (b) loading at least a portion of said biological fluid sample into a sample loading region of a vesicle capture device;
- (c) passing said portion of said biological fluid sample from said sample loading region through a vesicle-capture material disposed within said vesicle capture device, said vesicle-capture material comprising at least a first layer and a second layer, both of glassfiber,
- wherein said first layer is closer to said sample loading region than is said second layer,
- wherein said first layer has a porosity configured to capture material that is 1.6 microns or greater in diameter,
- wherein the second layer has a porosity configured to capture material that is about 0.6 microns to about 0.8 microns or greater in diameter,
- wherein said passing results in capture of said vesicles from said biological fluid sample on or in both of said first and second layers;
- (d) removing non-vesicle material from said device;
- (e) passing a lysis agent in a buffer from said sample loading region through said first layer and then subsequently through said second layer of said vesicle-capture material, thereby lysing said captured vesicles in or on both of said first and second layers with the lysis agent in the buffer to create a lysate comprising a first material from vesicles captured in or on said first layer, said lysate further comprising a second material from vesicles captured in or on said second layer;
- and
- (f) collecting the lysate from both of said first and second layers into a single receiving region, thereby isolating a biomarker from said vesicles.
A method of isolating a biomarker from biological fluid, comprising:
- (a) obtaining a biological fluid sample comprising vesicles comprising at least one biomarker;
- (b) loading a portion of said biological fluid sample into a well of a multi-well vesicle capture device;
- (c) passing said portion of said biological fluid sample through a vesicle-capture material disposed within said well, said vesicle-capture material comprising at least a first layer and a second layer, both of glassfiber,
- wherein said passing results in capture of said vesicles from said biological fluid sample on or in said first and second layers of said vesicle-capture material;
- wherein said first layer has a porosity configured to capture material that is 1.6 microns or greater in diameter,
- wherein the second layer has a porosity configured to capture material that is about 0.6 microns to about 0.8 microns or greater in diameter;
- (d) loading a portion of a lysis agent in a buffer onto said first layer;
- (e) passing said portion of said lysis agent in said buffer through the first layer and subsequently through the second layer to lyse said captured vesicles in or on said vesicle-capture material with said portion of said lysis agent in said buffer to create a lysate;
- and
- (f) collecting the lysate into a single receiving region, thereby isolating a biomarker from said vesicles captured in or on said vesicle-capture material disposed within said well.
Dependencies are inferred from the claim text.