Method and apparatus for the regulation of complex binding

• US 5,135,876 A
• Filed: 09/24/1987
• Issued: 08/04/1992
• Est. Priority Date: 09/24/1987
• Status: Expired due to Fees

• Current Assignee: University (290275) (Independent Bank Group Incorporated)
• Original Assignee: University of Utah (State of Utah)
• Inventors: Herron, James, Andrade, Joseph D.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): BEISNER, WILLIAM H
• Application Number:
  US07/100,935
• Time in Patent Office: 1,776 Days
• Field of Search: 436/527, 436/805, 436/806, 436/807, 436/824, 436/825, 436/512, 436/518, 436/524, 436/536, 436/164, 436/172, 422/57, 422/58, 422/68.1, 422/82.05, 422/82.08, 422/82.09, 422/82.11, 204/15, 204/402, 204/403, 385/12, 385/129-131, 250/362, 250/363.01, 250/458.1, 250/459.1, 250/472.1, 250/473.1
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10S 436/805 Optical property

Abstract

A reusable biosensor is disclosed. A molecule containing a moiety of an antibody-antigen complex and photosensitive polymers are bonded to an optical conduit. When placed in a solution, the presence or absence of a second complementary moiety of an antibody-antigen complex can be determined by sensing whether or not the first moiety is complexed. The first moiety of an antibody-antigen complex can then be regenerated by transmitting light through the optical conduit which alters the structure of the photosensitive polymers to cause interference with said complex, thereby dissociating the second complementary moiety from the first moiety. The ability to regulate and control specific binding has applications can be useful in information storage devices, bioorganic electronic devices, and optical devices.

Claims

1. A probe element capable of photon actuated chemical decoupling of a ligand comprising:

a conductive substrate constructed so as to conduct photons;

a plurality of first pendant molecules, each having a proximal end which is attached to said conductive substrate, and a distal end, wherein said distal end of each of said first pendant molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to a ligand having a chemical structure capable of coupling to said coupling group;

a plurality of second pendant molecules, each having a proximal end and a distal end, wherein said proximal end of each of said second pendant molecules is attached to said conductive substrate in proximity to at least one of said plurality of first pendant molecules, said second pendant molecules each having a first contracted conformation and a second extended conformation wherein said distal end of said second pendant molecules extends proximate to said coupling group of said first pendant molecules, said first contracted conformation existing when said second pendant molecules are in a substantially unenergized state and said second conformation existing when said second pendant molecules are subjected to a photon signal sufficient to extend said second pendant molecules from said first contracted conformation to said second extended conformation.

2. The probe element of claim 1 wherein said proximal end of each of said first pendant molecules is convalently bonded to said conductive substrate, and wherein said second extended conformation of said second pendant molecules causes dissociation of the ligand from said coupling group of said first pendant molecules.

3. The probe element of claim 2 wherein the photon signal is a wave of visble light and said second pendant molecules comprise photosensitive polymers.

4. The probe element of claim 1 wherein said conductive substrate comprises a planar waveguide.

5. The probe element of claim 1 wherein the coupling group of said first pendant molecules is a moiety of an antibody-antigen complex.

6. The probe element of claim 5 wherein said moiety of an antibody-antigen complex is an antibody.

7. The probe element of claim 5 wherein said moiety of an antibody-antigen complex is an antibody fragment.

8. The probe element of claim 5 wherein said moiety of an antigen-antibody complex is an antigen.

9. The probe element of claim 5 wherein said proximal end of each of said second pendant molecules is covalently bonded to said conductive substrate.

10. A method of externally regulating ligand-receptor binding comprising:

passing a solution containing ligands over a sensor comprising;

an optical conduit,

receptor molecules, each having a proximal end which is bonded to said optical conduit, and a distal end, wherein said distal end of each of said receptor molecules has a complexing site bonded thereto, said complexing site capable of complexing with said ligands, and

photoexpansive polymers bonded to said optical conduit and having free ends, interspersed among said receptor molecules, and having first and second conformations, wherein in said first conformation said free ends do not interfere with complexing of said ligand with said complexing site of said receptor molecules; and

transmitting light through said optical conduit to expand said photoexpansive polymers to said second conformation, wherein said free ends are extended proximate said complexing site of said receptor molecules to cause dissociation of said ligands from said receptor molecules.

11. An actuatable chemical coupling-decoupling device comprising:

a conductive substrate constructed so as to conduct a photon signal;

first pendant molecules each having a proximal end which is substantially immobilized upon said conductive substrate, and a free distal end, wherein said free distal end of each of said first pendant molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to chemical compounds having a complementary site capable of coupling to said coupling group of said first pendant molecules;

second pendant molecules each having a proximal and a distal end, wherein said proximal end of each of said second pendant molecules is attached to said conductive substrate adjacent to said first pendant molecules, and having a first contracted conformation wherein said distal end of said second pendant molecules is remote from said free distal end of said first pendant molecules and a second extended conformation wherein said distal end of said second pendant molecules extends into the vicinity of said free distal end of said first pendant molecules;

sensing means for sensing a change in condition produced by the coupling of said chemical compounds to said coupling group of said first pendant molecules;

photo signal means interacting with said conductive substrate to photoactivate said second pendant molecules to said extended conformation to cause decoupling of said chemical compounds from said coupling group of said first pendant molecules.

12. A reuseable biosensor system comprising:

an optical conduit;

binding molecules each having a proximal end which is attached to said optical conduit, and a distal end, wherein said distal end of said binding molecules has a moiety of an antibody-antigen complex attached thereto, said moiety pendantly extending from said optical conduit and capable of binding a complementary moiety to form said antibody-antigen complex;

photosensitive polymers each having a first end which is attach to said optical conduit at a point proximate said binding molecules, and a second end which is free, said photosensitive polymers having a first contracted unstimulated conformation existing when said photosensitive polymers are subjected to quanta of light in less than the amount required for stimulation, and a second extended conformation existing when a stimulating quanta of light is transmitted through said optical conduit, said photosensitive polymers being of sufficient length that when said second conformation exists, said second end of said photosensitive polymers extends proximate said moiety of an antibody-antigen complex at said distal end of said binding molecules;

a photon source constructed so as to transmit photons through said optical conduit and interacting with said optical conduit; and

sensing means operably disposed to interact with said optical conduit for sensing the presence or absence of said antigen-antibody complex on said binding molecules.

13. A probe element capable of photon actuated chemical coupling-decoupling of a ligand comprising:

a conductive substrate constructed so as to conduct a photon signal;

a plurality of first pendant molecules each having a proximal end which is attached to said conductive substrate, and a distal end, wherein said distal end of each of said first pendant molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to chemical compounds having a complementary site;

a plurality of second pendant molecules each having a proximal end which is attached to one of said first pendant molecules, and a distal end, said second pendant molecules having a first contracted conformation and a second extended conformation wherein said distal end of said second pendant molecules extends proximate to said coupling group of said first pendant molecules and interferes with binding of said chemical compounds to said coupling group, said first contracted conformation existing when no photon signal is conducted through said conductive substrate and said second conformation existing when said second pendant molecules are subjected to a photon signal to extend said second pendant molecules from said first contracted conformation to said second extended conformation.

14. The probe element of claim 13 wherein the photon signal is a wave of light and said second pendant molecules comprise photoexpansive polymers.

15. The probe element of claim 14 wherein said conductive substrate comprises a planar waveguide.

16. The probe element of claim 14 wherein the coupling group of said first pendant molecules is a moiety of an antibody-antigen complex.

17. The probe element of claim 16 wherein said moiety of an antibody-antigen complex is an antibody.

18. The probe element of claim 16 wherein said moiety of an antibody-antigen complex is an antibody fragment.

19. The probe element of claim 16 wherein said moiety of an antibody-antigen complex is an antigen.

20. The probe element of claim 13 wherein said first pendant molecules are antifluorescein antibodies.

21. A probe element capable of thermally-actuated chemical coupling-decoupling of a ligand comprising:

a conductive substrate constructed so as to conduct a thermal energy pulse;

a plurality of pendant hydrophilic molecules each having a proximal end which is attached to said conductive substrate, and a distal end; and

a plurality of pendant receptor molecules each having a proximal end which is attached to one of said pendant hydrophilic molecules, and a distal end, wherein said distal end of each of said pendant receptor molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to chemical compounds having a complementary coupling moiety capable of reversibly binding to said coupling group to form a complex, said complex being thermally sensitive to decouple said complementary coupling moiety from said coupling group when the thermal energy pulse is transmitted through said conductive substrate.

22. The probe element of claim 21 wherein said proximal end of each of said pendant hydrophilic molecules is covalently bonded to said conductive substrate.

23. The probe element of claim 22 wherein said thermal pulse is of a wave of near infrared light and said conductive substrate is an optical conduit capable of transmitting said wave of near infrared light.

24. The probe element of claim 22 wherein said conductive substrate is a heat conducting metallic strip.

25. The probe element of claim 23 wherein said coupling group of each of said pendant receptor molecules is a moiety of an antibody-antigen complex.

26. A probe element capable of thermally actuated decoupling of a ligand, comprising:

a conductive substrate constructed so as to conduct a thermal signal;

a plurality of pendant molecules each having a proximal end which is attached to said conductive substrate via a tethering compound, and a distal end, wherein said distal end of each of said pendant molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to chemical compounds having a complementary coupling moiety capable of reversibly binding to said coupling group to form a complex, said complex being thermally sensitive to decouple said complementary coupling moiety from said coupling group when the thermal signal is transmitted through said conductive substrate.

27. The probe element of claim 26, wherein said conductive substrate is further constructed to conduct a thermal signal of magnitude sufficient to cause said coupling group to reach a decoupling temperature between about 20°

C. and about 70°

C.

28. The probe element of claim 26, wherein said pendant molecules comprise moieties of an antigen-antibody complex.

29. A probe element capable of decoupling of a ligand in response to change in temperature of the element, comprising:

a conductive substrate constructed so as to conduct a thermal signal;

a plurality of pendant molecules each having a proximal end which is attached to said conductive substrate, and a distal end, wherein said distal end of each of said pendant molecules has a coupling group attached thereto, said coupling group constructed so as to selectively couple to chemical compounds having a complementary coupling moiety capable of reversibly binding to said coupling group to form a complex, said complex being thermally sensitive to decouple said complementary coupling moiety from said coupling group when the thermal signal is transmitted through said conductive substrate.

30. The probe element of claim 29 wherein said conductive substrate is an optical conductor and the thermal signal is infrared light.

31. The probe element of claim 30 wherein said pendant molecules are derived from antifluorescein antibodies.

32. The probe element of claim 29 wherein said conductive substrate is a metallic strip and the thermal signal is a resistive heat pulse.

33. The probe element of claim 32 wherein said pendant molecules comprise antifluorescein antibodies.