High density array fabrication and readout method for a fiber optic biosensor

• US 5,837,196 A
• Filed: 01/26/1996
• Issued: 11/17/1998
• Est. Priority Date: 01/26/1996
• Status: Expired due to Term

• Current Assignee: Medical Research Council (Government of United Kingdom), Regents of the University of California (University of California)
• Original Assignee: Medical Research Council (Government of United Kingdom), Regents of the University of California (University of California)
• Inventors: Segraves, Richard L., Zhai, Ye Yz, Pinkel, Daniel, Albertson, Donna G., Gray, Joe
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US08/592,779
• Time in Patent Office: 1,026 Days
• Field of Search: 455/5, 455/6, 455/91.2, 65/409, 65/55, 422/68.1, 422/82.05, 422/82.06, 422/82.07, 422/82.08, 422/82.09, 524/716
• US Class Current: 422/400
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 2201/0833 Fibre array at detector, resolving
• CPC Class Codes: G02B 6/04 formed by bundles of fibres G02B6/24 takes precedence

Abstract

The invention relates to the fabrication and use of biosensors comprising a plurality of optical fibers each fiber having attached to its "sensor end" biological "binding partners" (molecules that specifically bind other molecules to form a binding complex such as antibody-antigen, lectin-carbohydrate, nucleic acid-nucleic acid, biotin-avidin, etc.). The biosensor preferably bears two or more different species of biological binding partner. The sensor is fabricated by providing a plurality of groups of optical fibers. Each group is treated as a batch to attach a different species of biological binding partner to the sensor ends of the fibers comprising that bundle. Each fiber, or group of fibers within a bundle, may be uniquely identified so that the fibers, or group of fibers, when later combined in an array of different fibers, can be discretely addressed. Fibers or groups of fibers are then selected and discretely separated from different bundles. The discretely separated fibers are then combined at their sensor ends to produce a high density sensor array of fibers capable of assaying simultaneously the binding of components of a test sample to the various binding partners on the different fibers of the sensor array. The transmission ends of the optical fibers are then discretely addressed to detectors--such as a multiplicity of optical sensors. An optical signal, produced by binding of the binding partner to its substrate to form a binding complex, is conducted through the optical fiber or group of fibers to a detector for each discrete test. By examining the addressed transmission ends of fibers, or groups of fibers, the addressed transmission ends can transmit unique patterns assisting in rapid sample identification by the sensor.

Claims

1. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution, the process comprising the steps:

providing a matrix solution comprising mixing the biological binding partner and a solubilized matrix polymer dissolved in solution;

contacting the matrix solution with the solid surface, thereby attaching the biological binding partner such that it can specifically recognize and bind a second molecule.

2. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the step of forming the matrix solution comprises contacting an aqueous solution comprising the biological binding partner with a second solution comprising the matrix polymer.

3. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the biological binding partner is a nucleic acid and is attached to the solid surface such that the nucleic acid is capable of specifically hybridizing with a complementary nucleic acid.

4. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the solid surface is a glass slide.

5. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the solid surface is a sensor end of an optical fiber.

6. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the matrix polymer dissolved in solution is nitrocellulose.

7. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, wherein:

the matrix solution comprises nitrocellulose dissolved in DMSO.

8. A process for attaching a biological binding partner to a solid surface using a matrix polymer dissolved in solution according to claim 1, further comprising the step of drying the matrix solution after contacting the matrix solution with the solid surface.

22. The process for attaching a nucleic acid to a solid surface using a matrix polymer dissolved in solution according to claim 6, wherein the ratio of nitrocellulose to nucleic acid is between about 1:

10 and about 2;

1 by weight.

23. The process for attaching a nucleic acid to a solid surface using a matrix polymer dissolved in solution according to claim 22, wherein the ratio of nitrocellulose to nucleic acid is between about 1.5:

10 and about 1;

1 by weight.

24. The process for attaching a nucleic acid to a solid surface using a matrix polymer dissolved in solution according to claim 6, wherein the nitrocellulose solution is made by dissolving nitrocellulose in DMSO.

9. A process for constructing a fiber optic bundle for sensing a plurality of biological binding partners within a sample, the process comprising the steps of:

providing a plurality of optical fibers with each fiber having a sensor end and a transmission end wherein each fiber has attached to its sensor end a species of biological binding partner, the binding partner being attached to the sensor end of the optical fiber using a matrix polymer, such that the biological binding partner can specifically recognize and bind a second molecule;

combining said fibers with differing binding partners to form an optical fiber array wherein said fibers have commonly aligned sensor ends for simultaneous assay of a sample; and

,

addressing the transmission end of the combined discrete fibers for interrogation to produce the fiber optic sensor for the sample.

10. The process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the matrix polymer is nitrocellulose.

11. The process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the biological binding partner is a nucleic acid and is attached to the sensor end such that the nucleic acid is capable of hybridizing to a complementary nucleic acid.

12. The process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein said providing step comprises:

providing a plurality of optical fibers with each fiber having a sensor end and a transmission end;

placing the plurality of optical fibers in a plurality of fiber groups, each fiber group with commonly aligned sensor ends for simultaneous treatment;

providing a plurality of differing batches, each batch comprising a single species of biological binding partner suitable for attachment under treatment to the commonly aligned sensor ends of the optical fibers of a selected group of optical fibers;

placing and treating the commonly aligned sensor ends of differing fiber groups in differing batches with differing biological binding partners to produce a plurality of groups of optical fibers with the sensor ends of the optical fibers of each group having the same species of biological binding partner and the sensor ends of optical fibers of differing groups having differing species of biological binding partners; and

separating fibers from each group of fibers thereby providing a plurality of optical fibers with each fiber having a sensor end and a transmission end wherein each fiber has attached to its sensor end a species of biological binding partner.

13. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the combining separated fibers with differing binding partners with commonly aligned sensor ends includes randomly gathering the sensor ends.

14. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the combining separated fibers with differing binding partners with commonly aligned sensor ends includes assembling said sensor ends to form a tiered sensor face.

15. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the combining separated fibers with differing binding partners with commonly aligned sensor ends includes assembling said sensor ends to form a planer sensor face.

16. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the addressing the transmission end of the recombined discrete fibers for interrogation includes addressing the transmission ends of each of the discrete fibers to an optical array.

17. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

transmission ends of the optical fibers of each group having similar binding partners have similar markings for distinguishing the transmission ends corresponding to binding partners.

18. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 9, wherein:

the providing a plurality of batches of differing binding partners includes batches of nucleic acid binding partners to which nucleic acids in the sample might hybridize.

19. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 18, wherein:

the nucleic binding partners each correspond to specific regions on chromosomes.

20. A process for constructing a fiber optic bundle for sensing a plurality of binding partners within a sample according to claim 18, wherein:

the nucleic binding partners each correspond to expressed sequences.

21. A process for attaching a nucleic acid to a solid surface using a matrix polymer dissolved in solution, the process comprising the steps:

providing a matrix solution comprising dimethyl sulfoxide, nitrocellulose dissolved in solution and the nucleic acid; and

,

contacting the matrix solution with the solid surface, thereby attaching the nucleic acid such that it can specifically recognize and bind a second nucleic acid molecule.