Devices and methods for capturing target molecules
Provided herein are devices and methods for the capture or isolation of a biomarker from a biological sample. In several embodiments, the device comprises a loading region, a filter material, and a receiving region. In particular, in several embodiments, biological fluid is passed from the loading region through the filter material and into the receiving region, thereby resulting in capture or isolation of a biomarker.
1. A method of isolating vesicles from biological fluid, the method comprising:
(a) obtaining a urine sample comprising said vesicles;
(b) titrating said urine sample comprising said vesicles with a buffer solution comprising either phosphate buffered saline or HEPES to adjust the salt concentration of the urine sample comprising said vesicles to between 200 mM and 900 mM;
(c) adjusting the pH of the urine sample comprising said vesicles to between about pH 6 and pH 9; and
(d) passing said urine sample comprising said vesicles through a vesicle-capture material, said vesicle-capture material comprising glass-like materials, to produce a supernatant, wherein the vesicles from said urine sample are captured on or in said vesicle-capture material, wherein said titrating and said adjusting the pH of the urine sample comprising said vesicles is performed prior to passing said urine sample comprising said vesicles through the vesicle-capture material.
2. The method of claim 1 , wherein the supernatant is discarded.
3. The method of claim 1 , further comprising re-passing the supernatant through the vesicle-capture material.
4. The method of claim 1 , wherein said salt concentration is between 200 mM and 600 mM.
5. The method of claim 1 , wherein the salt concentration is based on the concentration of monovalent cations in the sample.
6. The method of claim 1 , wherein the salt concentration that is adjusted is the concentration of monovalent cations.
7. The method of claim 1 , wherein the said glass-like material comprises at least a first layer and a second layer of glass fiber.
8. The method of claim 7 , wherein the said glass-like material is configured to have a greater than 50% retention rate for particles having a diameter of from 0.6 microns to 1.5 microns in diameter.
9. The method of claim 1 , wherein the vesicles are selected from the group consisting of exosomes, vesicles, and other circulating membrane bound nucleic acid and/or protein-containing structures.
10. The method of claim 1 , wherein the vesicles contain a biomarker selected from the group consisting of mRNA, miRNA, rRNA, tRNA, and vRNA.
11. The method of claim 1 , wherein the salt concentration of the urine sample is adjusted to between about 200 mM and 600 mM.
12. The method of claim 1 , wherein said adjusting the pH of the urine sample is performed prior to passing said urine sample through the vesicle-capture material.
13. The method of claim 12 , wherein the salt concentration of the urine sample is adjusted to between about 200 mM and 600 mM.