Fluidic systems involving incubation of samples and/or reagents

Patent metadata

Field	Value
Publication number	US-10852310-B2
Application number	US-201615373056-A
Country	US
Kind code	B2
Filing date	Dec 8, 2016
Priority date	Dec 11, 2015
Publication date	Dec 1, 2020
Grant date	Dec 1, 2020

Abstract

Fluidic devices and methods involving incubation and/or mixing of assay components are provided. In some embodiments, a biological and/or chemical assay may be performed in a fluidic device. The fluidic device may be designed to allow for controlled incubation and/or mixing of two or more assay components. In some such embodiments, the fluidic device may comprise an incubation channel having a relatively large cross-sectional dimension in fluid communication with a detection channel. The incubation channel may allow for adequate mixing and/or incubation of two or more assay components prior to analysis of the assay. In some embodiments, fluidic devices for performing a vitamin D assay are provided.

First claim

What is claimed is:

1. A method, comprising:
introducing a sample into a fluidic device comprising at least one microfluidic channel, wherein the sample comprises vitamin D bound to a vitamin D binding protein;
flowing, at a first flow rate, at least a portion of the sample into a microfluidic incubation channel;
reducing the flow rate of the sample to a second flow rate after a leading edge of the sample enters the incubation channel but prior to the leading edge reaching an exit of the incubation channel, wherein the leading edge is a sample/gas interface, wherein the second flow rate is less than the first flow rate and/or is zero to allow incubation of the sample in the incubation channel, and wherein the incubation occurs for at least one minute at a temperature of at least 30° C.;
exposing the sample to a release agent;
releasing the vitamin D from the vitamin D binding protein;
modulating the flow rate of the sample to a third flow rate which is greater than the second flow rate; and
determining an amount of vitamin D in the sample.

2. A method as in claim 1 , wherein the fluidic device contains a buffer having a pH of at least 5 and less than or equal to 9.

3. A method as in claim 2 , wherein the buffer comprises a citrate buffer, an acetate buffer, a malonate buffer, a tris buffer, or a combination thereof.

4. A method as in claim 2 , wherein the buffer has an acidic pH.

5. A method as in claim 2 , wherein the buffer has a concentration of between 50 mM and 150 mM.

6. A method as in claim 1 , wherein the release agent comprises a detergent.

7. A method as in claim 6 , wherein the detergent comprises at least one of perfluorohexanoic acid, FS50, FS51, and Empigen BB.

8. A method as in claim 6 , wherein the detergent comprises at least one of 1-5% perfluorohexanoic acid, 0.4-0.8% FS50, 0.4% FS51, and 0.4% Empigen BB.

9. A method as in claim 1 , wherein the fluidic device contains in a channel a labeled molecule, wherein the labelled molecule is a monoclonal or polyclonal antibody that binds with vitamin D.

10. A method as in claim 1 , wherein the fluidic device contains, in a channel, a labeled molecule, wherein the labelled molecule is Vitamin D that binds with anti-Vitamin D antibody and/or fragments thereof.

11. A method as in claim 1 , wherein the fluidic device contains, in a channel, a labeled molecule, and wherein the labeled molecule is a metal nanoparticle-labeled anti-vitamin D antibody, a metal nanoparticle-labeled anti-vitamin D antibody fragment, a gold-labeled anti-vitamin D antibody, or a gold-labeled anti-vitamin D antibody fragment.

12. A method as in claim 1 , wherein the fluidic device contains, in a channel, a labeled molecule, and wherein the labeled molecule is a metal nanoparticle-labeled Vitamin D, or a gold-labeled Vitamin D.

13. A method as in claim 1 comprising diluting the sample between 0 vol % to 90 vol %.

14. A method as in claim 1 , wherein the sample comprises whole blood.

15. A method as in claim 1 , wherein the sample is serum or plasma.

16. A method as in claim 1 , wherein the release agent is a dried or lyophilized solid derived from a liquid solution.

17. A method as in claim 1 , wherein the release agent is a liquid solution.

18. A method as in claim 17 , wherein the liquid solution further contains a transfer molecule.

19. A method as in claim 18 , wherein the transfer molecule is a member of the beta-cyclodextrin family.

20. A method as in claim 18 , wherein the transfer molecule is methyl-beta-cyclodextrin, or comprises an acetyl-, succinyl-(2-hydroxypropyl)-, 2-hydroxypropyl-, carboxymethyl-, sulfate, 2-hydroxyethyl, succinyl-, and/or butyl- group.

21. A method as in claim 1 , wherein the release agent comprises a carboxyl or amine oxide head group and a fluorinated or non-fluorinated carbon chain.

22. A method as in claim 1 , wherein the dried or lyophilized release agent contained in the fluidic channel is reconstituted by the sample.

23. A method as in claim 1 , wherein the release agent is contained in a first fluid and the anti-vit-D antibody and/or fragments thereof is contained in a second fluid or the Vitamin D is contained in a second fluid, and wherein the second fluid is separated from the first fluid by a third fluid that is immiscible with the first and second fluids.

24. A method as in claim 1 comprising heating the sample to at least about 50° C. for at least 5 minutes and less than or equal to 15 minutes.

25. A method as in claim 1 , wherein the steps of exposing the sample to a release agent and releasing the vitamin D from the vitamin D binding protein occur before or during the step of flowing, at the first flow rate, at least a portion of the sample into an incubation channel.

26. A method as in claim 1 , wherein the steps of exposing the sample to a release agent and releasing the vitamin D from the vitamin D binding protein occur after the step of flowing, at the first flow rate, at least a portion of the sample into an incubation channel.

27. A method as in claim 1 , further comprising binding the released vitamin D with a labelled anti-vit-D antibody, wherein a plurality of the labelled anti-vit-D antibodies are bound to released vitamin D or are bound to vitamin D immobilized on a surface of the at least one microfluidic channel, the method further comprising flowing a wash plug comprising a rinse solution past the plurality of labelled anti-vit-D antibodies, wherein the wash plug is separated from the sample by a fluid that is immiscible with both the sample and the rinse solution.

28. A method as in claim 1 , further comprising binding the released vitamin D and/or labelled vitamin D with a plurality of anti-vit-D antibodies immobilized on a surface of the at least one microchannel, the method further comprising flowing a wash plug comprising a rinse solution past the plurality of anti-vit-D antibodies immobilized on the surface of the at least one microchannel, wherein the wash plug is separated from the sample by a fluid that is immiscible with both the sample and the rinse solution.

29. A method as in claim 1 , wherein the step of exposing the sample to the release agent occurs before or during the step of introducing the sample into the fluidic device.

30. A method as in claim 1 , wherein the step of exposing the sample to the release agent occurs after the step of introducing the sample into the fluidic device.

31. A method as in claim 1 , wherein the method further comprises:
exposing the sample to a gold-labeled molecule; and
exposing the gold-labeled molecule to a reagent after the step of exposing the sample to the gold-labeled molecule,
wherein the step of determining the amount of vitamin D in the sample is based at least in part on a signal generated by a chemical reaction between the gold-labeled molecule with the reagent.

32. A method as in claim 31 , wherein the chemical reaction between the gold-labeled molecule and the reagent occurs in a microfluidic channel.

33. A method, comprising:
exposing a sample comprising vitamin D bound to a vitamin D binding protein to a release agent;
releasing the vitamin D from the vitamin D binding protein;
introducing the sample subjected to the exposing step into a fluidic device comprising at least one microfluidic channe

Assignees

• Opko Diagnostics Llc

Inventors

• Haddad Douglas M
• Fagan Gary J
• Dong Yi
• Dirckx Matthew
• Linder Vincent

Classifications

• B01L3/502707

  characterised by the manufacture of the container or its components · CPC title

• B01L2300/0874

  Three dimensional network · CPC title

• B01L3/502784

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title

• B01L2300/0867

  Multiple inlets and one sample wells, e.g. mixing, dilution · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title