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Electronically-indexed solid-phase assay for biomolecules    
United States Patent5641634   
Link to this pagehttp://www.wikipatents.com/5641634.html
Inventor(s)Mandecki; Wlodek (Libertyville, IL)
AbstractDisclosed are materials and methods for detecting biomolecules in samples employing transponders having memory elements associated with particle(s) used as a solid phase in art assay, and information pertinent to the assay is encoded on the transponder memory elements. A dedicated read/write device is used remotely to encode or remotely to read the information encoded on the transponder memory elements. The invention can be used in direct or competitive ELISA-type assays, or in multiplex assays for the simultaneous assay of several analytes.
   














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Drawing from US Patent 5641634
Electronically-indexed solid-phase assay for biomolecules - US Patent 5641634 Drawing
Electronically-indexed solid-phase assay for biomolecules
Inventor     Mandecki; Wlodek (Libertyville, IL)
Owner/Assignee    
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Publication Date     June 24, 1997
Application Number     08/565,007
PAIR File History     Application Data   Transaction History
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Litigation
Filing Date     November 30, 1995
US Classification    
Int'l Classification    
Examiner     Chin; Christopher L.
Assistant Examiner     Nguyen; Bao-Thuy L.
Attorney/Law Firm     Brinks Hofer Gilson & Lione
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Patent Tags     electronically-indexed solid-phase assay biomolecules
   
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5491097
Ribi
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Holm-Kennedy
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 Technical Review Submit all comments and votes
 Claims Submit all comments and votes
 


I claim:

1. A solid phase for use in assays for biomolecules, comprising:

(a) a particle and a transponder, wherein the transponder is located inside said particle or is attached to the surface of said particle, the transponder comprising a radio transmitter-receiver encoded with data, the data is transmitted to a receiver in response to a specific dectromagnetic signal; and

(b) a member of a biomolecular binding pair attached to the surface of the pattide, wherein the biomolecule specifically binds to the biomolecular binding pair member.

2. The particle of claim 1, wherein the surface of the particle is glass, latex or plastic.

3. The particle of claim 1, wherein the biomolecular binding pair is an antigen-antibody pair.

4. A method of detecting a member ofa biomolecular binding pair in a sample, comprising the steps of:

(a) providing at least one solid phase particle, said particles having a transponder located inside or attached to each of said particles, the transponders having memory dement and an index number encoded on the memory elements creating at least one class of transponders, each class having a different index number;

(b) the solid particles having a first member of said biomolecular binding pair attached to a surface thereof;

(c) contacting the solid phase particles with a sample to cause a second member of the biomolecular binding pair to specifically bind to the first member attached to the solid phase particle;

(d) adding a label reagent to the sample, said label reagent specifically binds the second member of the biomolecular binding pair on the solid phase particle, and analyzing the solid phase particles to detect the presence of the label indicative of binding of the second member present in the sample to said first member; and

(e) decoding the index number encoded on the transponders using a scanner device to identify the class of the transponders to which the second member of the biomolecular binding pair is bound.

5. The method of claim 4, wherein the index number is encoded on the transponder memory element by the transponder manufacturer.

6. The method of claim 4, wherein the index number is encoded on the transponder memory element by the user with a scanner device.

7. The method of claim 4 wherein the label is a chromophore.

8. The method of claim 4 wherein the label is a fluorophore.

9. The method of claim 4, wherein the label is a chemiluminescent agent or a bioluminescent agent.

10. The method of claim 4, wherein an outer surface of the transponders is glass, plastic or latex.

11. The method of claim 4 wherein the index number comprises physical or chemical characteristics of the biomolecular binding pair member deposited on the solid phase.

12. The method of claim 4 wherein the index number comprises identifying characteristics of the sample.

13. A method of detecting at least two biomolecules in a sample, said biomolecules being a second member of a biomolecular binding pair, comprising the steps of:

(a) introducing into the sample at least two populations of solid phase particle, said particles having a transponder located inside or attached to each of said particles, each population of particle having a first member of a biomolecular binding pair attached to its surface, and the transponders in the first population being encoded with a different index number than the transponders of the second population, wherein, the first member on each population of particles specifically binds one of said second members present in said sample;

(b) contacting the solid phase with a label reagent that specifically binds to the second member of the biomolecular binding pair, and analyzing the particles to detect a label indicating binding of the second biomolecular binding pair member to the first biomolecular binding pair member; and

(c) decoding at least a portion of the transponders to determine the populations of the transponders to which the second member is bound.

14. The method of claim 13, wherein said biomolecules comprise nucleic acid sequences and the solid phase particles comprise at least three populations of solid phase particles, each particle comprising a transponder and each particle having an oligonucleotide attached to a surface of the particle, each of the three populations having a different oligonucleotide sequence attached to the particle and each of the populations of particles being encoded with a different index number.

15. A method of performing a multiplex solid phase assay for biomolecules in a sample, said biomolecules being a second member of a biomolecular binding pair comprising the steps of:

(a) providing multiple solid phases comprising particles, said particles having a transponder located inside or attached to each of said particles, the transponders having memory elements encoded with an index number creating two or more classes of transponders, each class having a different index number, each class of solid phase particles having a first biomolecular binding member immobilized on a surface thereof, wherein, the first member on each class of particles specifically binds one of said second members present ill said sample:

(b) contacting the solid phase particles with a sample to cause two or more different second members of a biomolecular binding pair in the sample to bind one of said first biomolecular binding pair member on the particles;

(c) contacting the solid phase particles with label reagents that specifically bind to the second member of the biomolecular binding pair;

(d) washing the solid phase particles to remove unbound sample components;

(e) analyzing the solid phase particles to detect the label, which indicates binding of the second biomolecular binding pair member to the first biomolecular binding pair member; and

(f) decoding the index number encoded on at least a portion of the transponders to identify the class of transponders to which a biomolecule is bound.

16. A kit for detecting the presence of a member of a biomolecular binding pair in a sample, comprising:

(a) at least one assay vessel, containing at least one solid phase particle, the particle comprising a transponder, the transponder having a memory element and the particle having a primary layer of biomolecules bound to a surface of the particle;

(b) at least one probe reagent, comprising one member of the biomolecular binding pair; and

(c) at least one label reagent that binds selectively to the probe reagent.

17. The kit of claim 16, further comprising:

(a) at least one positive control, comprising a solution of a member the biomolecular binding pair; and

(b) at least one negative control, comprising a solution free of the biomolecular binding pair member.

18. The kit of claim 17, further comprising a sample diluent buffer solution.

19. The kit of claim 16, wherein the primary layer comprises protein antigens.

20. The kit of claim 16, wherein the primary layer of biomolecules comprise vital antigens.

21. The kit of claim 16, wherein the biomolecular binding pair member to be detected comprises a cell.

22. A method of detecting a member of a biomolecular binding pair in a sample comprising the steps of:

(a) providing at least one solid phase particle, said particles having a transponder located inside or attached to each of said particles, the transponders having memory elements and an index number encoded on the memory elements creating at least one class of transponders, each class having a different index number;

(b) the solid particles having a first member of said biomolecular binding pair attached to the surface of the solid phase particles;

(c) contacting the solid phase particles with a sample and a label reagent to cause a second member of the biomolecular binding pair present in said sample, and the label reagent to competitively bind to the first member attached to the solid phase particle;

(d) analyzing and detecting the presence of the label on the solid phase particle as an indirect determination of the presence of the second member in said sample;

(e) decoding the index number encoded on the transponders using a scanner device to identify the class of transponders to which the second member of the biomolecular binding pair is bound.

23. A method of detecting at least two biomolecules in a sample, said biomolecules being a second member of a biomolecular binding pair, comprising the steps of:

(a) providing at least two populations of solid phases comprising particles, said particles having a transponder located inside or attached to each of said particles, each population of particle having a first member of a biomolecular binding pair attached to its surface, and the transponders in the first population being encoded with a different index number than the transponders of the second population, wherein, the second members present in said sample competitively binds one of said first member on each population of the solid phase particles;

(b) contacting the solid phase particles with a sample and label reagents to cause the second members present in said sample, and the label reagents to competitively bind to one of said first member on the solid phase particles;

(c) analyzing the particles to detect a label to indirectly determine the presence of the second biomolecular binding pair member in the sample;

(d) decoding at least a portion of the transponders to determine the populations of the transponders.

24. A method of performing a multiplex solid phase assay for biomolecules in a sample, said biomolecules being a second member of a biomolecular binding pair, comprising the steps of:

(a) providing multiple solid phases comprising particles, said particles having a transponder located inside or attached to each of said particles, the transponders having memory elements encoded with an index number creating two or more classes of transponders, each class having a different index number, each class of solid phase particles having a first biomolecular binding member immobilized on a surface thereof, wherein, the second members present in said sample competitively bind to one of said first member on the multiple solid phase particles;

(b) contacting the solid phase particle with a sample and label reagents to cause the second members and the label reagents to competitively bind to one of said first member on the multiple solid phase particles;

(d) washing the solid phase to remove unbound sample components;

(e) analyzing the solid phase to detect the label, which indirectly indicates binding of the second biomolecular binding pair member to the first biomolecular binding pair member; and

(f) decoding the index number encoded on at least a portion of the transponders to identify the class of transponders to which a biomolecule is bound.
 Description Submit all comments and votes
 


BACKGROUND OF THE INVENTION

This invention relates to materials and methods for detecting biomolecules in samples, and more particularly to a particulate solid phase having for encoding information concerning the assay, and to assays employing such a solid phase.

Solid phase assays have been used to determine the presence and/or the concentration of biomolecules, such as proteins, peptides, nucleic acids, carbohydrates and lipids. Solid-phase assays can be performed in a variety of fluids, e.g., simple buffers, biological fluids, such as blood, serum, plasma, saliva, urine, tissue homogenates, and many others.

In solid phase assays, small beads, or microparticles, are typically used as the solid phase to capture the analyte. Solid phase microparticles can be made of a variety of materials, such as glass, plastic or latex, depending on the particular application. Some solid phase particles are made of ferromagnetic materials to facilitate their separation from complex suspensions or mixtures.

In conventional solid-phase assays, the solid phase mainly aids in separating biomolecules that bind to the solid phase from molecules that do not bind to the solid phase. Separation can be facilitated by gravity, centrifugation, filtration, magnetism, immobilization of molecules onto the surface of the vessel, etc. The separation may be performed either in a single step in the assay or, more often, in multiple steps.

Often, it is desirable to perform two or more different assays on the same sample, in a single vessel and at about the same time. Such assays are known in the art as multiplex assays. Multiplex assays are performed to determine simultaneously the presence or concentration of more than one molecule in the sample being analyzed, or alternatively, to evaluate several characteristics of a single molecule, such as, the presence of several epitopes on a single protein molecule.

One problem with conventional multiplex assays is that they typically cannot detect more than about five analytes simultaneously, because of difficulties with simultaneous detection and differentiation of more than about five analytes. In other words, the number of different analytes that may be assayed simultaneously is limited by the solid phase.

SUMMARY OF THE INVENTION

This invention overcomes many of these problems by the use of transponders associated with the solid phase beads to index the particles constituting the solid phase. Thus, each individual transponder-containing solid phase particle can be assigned a unique index number, electronically encoded inside the particle, that can be retrieved by the scanner device at any time, e.g., at one time during the assay, at multiple times during the assay, or continuously during the assay. The index number may relate to the time and date on which the assay was performed, the patient's name, a code identifying the type of the assay, catalog numbers of reagents used in the assay, or data describing the progress of the assay, such as temperature during different steps of the assay.

In an electronically-indexed multiplex assay of this invention, two or more transponders, each encoded with a different index number and constructed to bind a different analyte, are incubated with the sample in a single vessel. After necessary additions, incubations and washes are performed, which are similar to incubations and washes in existing assays, the solid phase is analyzed to detect a label indicative of binding of the analyte to the solid phase, such as fluorescence, color, radioactivity or the like. Solid phase analysis is either preceded or followed by the decoding of the index number on the transponder.

Determination of the label and decoding of the memory of the transponder can be done manually on two different instruments, such as a fluorometer and a dedicated scanner, although a single automated instrument that would perform both functions may be used. Such an instrument can be a modified fluorometer in which the scanner is mounted in the proximity of the fluorometer readout window, and reading the sample fluorescence and decoding the transponder are coordinated by a central computer. In addition, such an instrument can be equipped with an automated transport system for transponders.

In one aspect, the present invention provides an electronically-indexed solid phase particle for use in solid phase assays for biomolecules, comprising a transponder and a member of a biomolecular binding pair attached to the transponder.

In another aspect, the present invention provides a method of detecting biomolecules in a sample using solid phase particles having transponders.

In another aspect, the present invention includes a kit for detecting biomolecules in a sample using transponders, comprising assay vessels, a probe reagent, and a labelled conjugate reagent.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic representation of a simple assay of this invention.

FIG. 2 is a schematic representation of a multiplex assay of this invention.

FIG. 3 is a cross-sectional view of a solid phase particle with a transponder and a primary layer of biomolecules bound to a surface thereof.

FIG. 4 is a schematic diagram of the signal pathway for encoding and decoding data on the transponders.

FIG. 5 is a schematic representation of a miniature transponder.

FIG. 6 is a plan view of a miniature transponder.

FIG. 7 is a plan view of a transport system/analytical instrument for implementing the present invention.

FIG. 8 is a plan view of a modified flow cytometer for high speed analysis of solid phase particles of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

FIG. 1 depicts a simple assay of the invention. A solid phase particle 10, with a transponder 12 is derivatized by attaching an antibody 11 to the outer surface 16 of the particle 10. Information concerning the assay, e.g., the assay lot number, is encoded on the transponder, either by the manufacturer of the tra