Method of manufacturing a plurality of uniform microfabricated sensing devices having an immobilized ligand receptor

• US 5,063,081 A
• Filed: 08/15/1990
• Issued: 11/05/1991
• Est. Priority Date: 11/14/1988
• Status: Expired due to Term

• Current Assignee: i STAT Corporation (Abbott Laboratories Incorporated)
• Original Assignee: i STAT Corporation (Abbott Laboratories Incorporated)
• Inventors: Cozzette, Stephen N., Mier, Randall M., Itak, Jeanne, Davis, Graham, Lauks, Imants R., Wieck, Henry J., Smit, Nicolaas, Piznik, Sylvia, Van Der Werf, Paul, Steiner, Susan
• Primary Examiner(s): Beck, Shrive
• Assistant Examiner(s): Owens, Terry J.
• Application Number:
  US07/567,870
• Time in Patent Office: 447 Days
• Field of Search: 427/2, 427/407.1, 427/414, 204/403, 204/415, 204/416, 204/418, 204/153.12, 422/57, 422/58, 435/7.1, 435/299, 435/817, 436/518, 436/805, 436/810
• US Class Current: 435/4
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 3/0268 using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: G01N 2035/00059 vacuum chucks
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/00455 Controlling humidity in analyser
• CPC Class Codes: G01N 2035/00534 Mixing by a special element, e.g. stirrer
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• CPC Class Codes: G01N 2035/1076 plurality or independently movable heads
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/109 with two horizontal degrees of freedom

Abstract

A plurality of uniform microfabricated sensing devices are produced by establishing a plurality of base sensors on a substrate wafer, forming over at least a portion of each base sensor a permselective layer, superimposing a photoformable proteinaceous photoresist layer over a substantial portion of the permselective layer, and forming a topmost layer of an immobilized ligand receptor. The ligand receptor and corresponding ligand may be immunoreactive species.

Claims

1. A method of manufacturing a plurality of uniform microfabricated sensing devices which comprises:

(a) establishing a plurality of base sensors on a suitable substrate wafer;

(b) establishing a permselective layer, superimposed over at least a portion of each base sensor, having a thickness sufficient to exclude substantially molecules with a molecular weight of about 120 or more while allowing the free permeation of molecules with a molecular weight of about 50 or less;

(c) establishing a photoresist layer comprising a photoformable proteinaceous mixture, superimposed over a substantial portion of said permselective layer; and

(d) establishing a topmost layer comprising a sufficient amount of an immobilized ligand receptor.

2. The method of claim 1 which further comprises establishing an electrolyte layer over at least a portion of each base sensor, prior to establishing said permselective layer.

3. The method of claim 1 in which said ligand receptor is an immunoreactive species.

4. The method of claim 1 in which said ligand receptor is selected from the group consisting of ionophores, cofactors, polypeptides, proteins, glycoproteins, enzymes, immunoglobulins, antibodies, antigens, lectins, neurochemical receptors, oligonucleotides, polynucleotides, molecules of DNA, molecules of RNA, active fragments or subunits or single strands of the preceding molecules, and mixtures thereof.

5. The method of claim 1 in which said ligand receptor is an antibody.

6. The method of claim 1 in which said ligand receptor is an antigen.

7. The method of claim 1 in which said ligand receptor is a polypeptide.

8. The method of claim 1 in which said ligand receptor is an oligonucleotide.

9. The method of claim 1 in which said ligand receptor is immobilized over a preselected area of each base sensor.

10. The method of claim 1 in which each base sensor comprises an indicator electrode and said ligand receptor is immobilized over the indicator electrode of each base sensor.

11. The method of claim 1 in which said ligand receptor is immobilized by covalent attachment.

12. The method of claim 1 in which said ligand receptor is immobilized by absorption.

13. The method of claim 1 in which said permselective layer comprises a heat-treated film of a silane compound having the formula R'"'"'_n Si(OR)_4-n, in which n is an integer selected from the group consisting of 0, 1 and 2;

R'"'"' is a hydrocarbon radical comprising 3-12 carbon atoms; and

R is a hydrogen radical or a lower alkyl radical comprising 1-4 carbon atoms.

14. The method of claim 13 in which said silane compound is N-(2-aminoethyl)-3-aminopropyltrimethoxysilane.

15. The method of claim 1 in which said permselective layer comprises a copolymer of a siloxane compound and a nonsiloxane compound.

16. The method of claim 15 in which said copolymer is selected from the group consisting of dimethylsiloxane-alkene oxide, tetramethyldisiloxane-divinylbenzene, tetramethyldisiloxane-ethylene, dimethylsiloxane-silphenylene, dimethylsiloxane-silphenylene oxide, dimethylsiloxane, methylstyrene, and dimethylsiloxane-bisphenol A carbonate, and mixtures thereof.

17. The method of claim 15 in which said copolymer is dimethylsiloxane-bisphenol A carbonate.

18. The method of claim 11 in which said covalent attachment is accomplished with the benefit of a crosslinking agent.

19. The method of claim 18 in which said crosslinking agent is a polyfunctional compound having at least two functional groups selected from the group consisting of formyl, vinyl, carboxyl, anhydride, amine, amide, epoxy, hydroxyl, cyano, isocyanato, thio, halo and stable combinations thereof.

20. The method of claim 1 in which said photoformable proteinaceous mixture comprises:

(i) a proteinaceous substance;

(ii) an effective amount of a photosensitizer uniformly dispersed in said proteinaceous substance; and

(iii) water.

21. The method of claim 20 in which said proteinaceous substance is fish gelatin.

22. The method of claim 20 in which said photosensitizer is a high oxidation state transition metal compound.

23. The method of claim 20 in which said photoformable proteinaceous mixture further comprises a crosslinking agent.

24. The method of claim 1 or 2 in which each base sensor comprises an electrochemical transducer.

25. The method of claim 24 in which said electrochemical transducer is amperometric.

26. The method of claim 24 in which said electrochemical transducer is potentiometric.

27. The method of claim 1 or 2 in which each base sensor comprises an amperometric electrochemical transducer comprising an indicator electrode which includes an electrocatalyst selected from the group consisting of carbon, platinum, gold, silver, rhodium, iridium, ruthenium, mercury, palladium, and osmium.

28. The method of claim 1 or 2 in which each base sensor comprises a potentiometric electrochemical transducer comprising an indicator electrode which includes a mixed metal oxide alloy electrocatalyst.

29. The method of claim 1 or 2 in which each base sensor comprises a potentiometric electrochemical transducer comprising an indicator electrode which includes a silver/silver halide electrocatalyst.

30. The method of claim 24 in which said electrochemical transducer further comprises a reference electrode which includes an electrocatalyst metal selected from the group consisting of silver, gold, and platinum.

31. The method of claim 24 in which said electrochemical transducer further comprises a silver/silver halide reference electrode.