Claims
- 1. A method for detecting biological substances present in a conductive liquid medium (LC), comprising:
- providing at least one affinity sensor which includes at least one structure having at least one semiconductor material coated on one surface with at least one isolating layer and at least one sensitive membrane affixed to said isolating layer, said sensitive membrane in fluid contact with a conductive liquid medium (LC) and which includes at least one ligand that is complementary to and capable of forming a hybridizing pair with said biological substance;
- applying a voltage between the semiconductor and the conductive liquid medium;
- detecting variations in electrical signals resulting from a pairing of said biological substance in said conductive liquid medium with said ligand coupled on said sensitive membrane; and
- interpreting the variations in electrical signals detected in said detecting step in order to detect said biological substance.
- 2. The method of claim 1, wherein:
- said sensor further comprises at least one working electrode consisting of the semiconductor/isolating layer/sensitive membrane structure, at least one reference electrode that forms a reference system and at least one counter-electrode;
- said applying step comprises applying a polarization voltage V.sub.pol to said working electrode, said polarization voltage being continuous in relation to said reference electrode immersed in said conductive liquid medium, superimposing on said voltage V.sub.pol, a sinusoidal electrical modulation; and
- said detecting step comprises measuring a voltage between the working electrode and the reference electrode, and a current passing through the working electrode, calculating a phase impedance Zp or the quadrature impedance Zq, plotting the Zp or Zq curve as a function of V.sub.pol, and monitoring a shift of said curve, with said shift corresponding to said variations in the electrical signals, as induced by said charge effect resulting from said pairing of said biological substance with said ligand.
- 3. The method of claim 1 or 2, wherein said pairing comprises a reaction selected from the group consisting of nucleotide hybridization, and immunological coupling.
- 4. The method of claim 2, wherein said shift in said Zp or Zq curve is monitored by monitoring a flat-band potential (V.sub.bp) of said semiconductor material, which is deducted from the said curve.
- 5. The method of claim 2, wherein said polarization voltage varies within a depletion range of said semiconductor material.
- 6. The method of claim 2, wherein said polarization voltage varies within a depletion range of said semiconductor material.
- 7. The method of claim 2, wherein said polarization voltage is from -3 to +3 V.
- 8. The method of claim 2, wherein said polarization voltage is from -1.5 to +1.5 V.
- 9. The method of claim 1, wherein:
- said affinity sensor includes at least one transducer element formed by at least one field-effect transistor having a gate in contact with the conductive liquid medium, said gate formed by said sensitive membrane, with said transistor being associated with at least one reference electrode that is also immersed in said conductive liquid medium; and
- said electrical signals induced by said pairing of said biological substance with said ligand on said sensitive membrane comprise variations in current or voltage.
- 10. The method of claim 9, wherein:
- a potential (V.sub.ds) between a drain and a source of said polarization voltage (V.sub.pol) is assigned a constant value;
- a drain current is kept constant, at an index value, with a potential between the reference and the source, V.sub.rs, being adjusted; and
- variations in V.sub.rs are measured in response to said pairings between the biological substances and said ligands on said sensitive membrane.
- 11. The method of claim 1, wherein said affinity sensor further comprises at least one semiconductor/isolating layer structure which does not include a sensitive membrane and in which the isolating layer and the semiconductor form a field-effect transistor (FET) used as a reference (REFET).
Priority Claims (1)
Number |
Date |
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2 722 294 |
Jul 1994 |
FRX |
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CROSS REFERENCE TO RELATED APPLICATIONS
This application is a divisional application of and claims the benefit of U.S. patent application Ser. No. 08/649,985, filed on May 17, 1996, now U.S. Pat. No. 5,869,244 the disclosure of which is hereby incorporated by reference in its entirety for all purposes.
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EPX |
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Divisions (1)
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Number |
Date |
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Parent |
649985 |
May 1996 |
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