Physiologic sample preparation for nanosensors
The present invention provides a microfluidic purification chip for capturing a biomarker from a physiological solution. The present invention also provides a method of capturing and releasing a biomarker, wherein the biomarker is originally in a physiological solution. The present invention further provides a method of pre-purifying and measuring the concentration of a biomarker in a physiological solution.
1. A method of pre-purifying and measuring the concentration of at least one biomarker in a physiological solution, wherein the method comprises the steps of:
contacting the physiological solution comprising the at least one biomarker with a microfluidic purification chip,
wherein the microfluidic purification chip comprises a first antibody directed to binding of a first epitope of the at least one biomarker, wherein the first antibody is attached to the microfluidic purification chip by a molecular crosslinker, wherein the first antibody is conjugated to the molecular crosslinker, and the molecular crosslinker is immobilized on the microfluidic purification chip,
wherein the molecular crosslinker comprises a molecular spacer and a cleavable group;
removing the physiological solution from the microfluidic purification chip;
optionally washing the microfluidic purification chip with a buffer;
cleaving the cleavable group in a sensing buffer to generate a biomarker-containing solution comprising complexes of the at least one biomarker bound to the first antibody;
transferring the biomarker-containing solution to a sensing chip, wherein the sensing chip comprises a nanowire sensor and the nanowire sensor is derivatized with a second antibody directed to binding of a second different epitope of the at least one biomarker of the complexes, wherein binding of the first antibody to the first epitope of the at least one biomarker does not prevent binding of the second antibody to the second epitope of the at least one biomarker;
contacting the biomarker-containing solution with the sensing chip wherein the second antibody binds the second epitope of the at least one biomarker, bound at the first epitope to the first antibody, of the complexes; and determining the concentration of the at least one biomarker, bound to the first antibody and the second antibody, in the biomarker-containing solution.
2. The method of claim 1 , wherein the microfluidic purification chip is connected to the sensing chip by a duct, wherein the duct is used to transfer the biomarker-containing solution from the microfluidic purification chip to the sensing chip, the duct optionally comprising a valve.
3. The method of claim 1 , wherein the sensing chip comprises a nanoribbon sensor.
4. The method of claim 1 , wherein the sensing chip produces a gate voltage which ranges from about −2.5 V to about −6 V.
5. The method of claim 1 , wherein the sensing solution has a Debye screening length of about λ D =9.6 nm.
6. The method of claim 1 , wherein the cleavable group is cleaved with UV or visible light or is cleaved with an acid, a base, an oxidant, or a reducer.
7. The method of claim 1 , wherein the cleavable group of the molecular crosslinker is coupled to a biotin-containing moiety.
8. The method of claim 7 , wherein the microfluidic purification chip is derivatized with avidin and the biotin-containing moiety of the molecular crosslinker is immobilized on the microfluidic purification chip via a binding interaction between the biotin-containing moiety and the avidin.
9. The method of claim 1 , wherein the physiological solution is whole blood.
10. The method of claim 1 , wherein the physiological solution comprises two or more biomarkers.
11. The method of claim 1 , wherein the molecular spacer is selected from the group consisting of a peptide, a nucleic acid, a polyethylene glycol, and an alkylene group.