Method and apparatus for detecting estradiol and metabolites thereof using an acoustic device
First Claim
1. A method for detecting estradiol in a sample, the method comprising the steps of:
- a) introducing a fluid comprising a sample and a plurality of magnetic particles coated with a first capture agent capable of binding estradiol into a fluid chamber, wherein at least one surface of the fluid chamber comprises a flexural plate wave device having a second capture agent capable of binding estradiol, wherein the flexural plate wave device has a membrane that is capable of vibrating, and wherein the second capture agent is bound to the membrane;
b) applying a retractable source of magnetic flux positioned external to the fluid chamber close to the membrane to create a significant magnetic field gradient at a surface of the membrane to attract magnetic particles in the fluid toward the membrane;
c) monitoring a first signal output by said flexural plate wave device, wherein the first signal output is monitored in the presence of a magnetic flux;
d) after removing the retractable source of magnetic flux, flowing a solution through the fluid chamber to remove magnetic particles not bound to the membrane;
e) monitoring a second signal output by said flexural plate wave device, wherein the second signal output is monitored in the absence of the magnetic flux; and
f) comparing the first and the second signal output to a control signal, thereby detecting estradiol in the sample.
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Abstract
Methods for detecting estradiol and metabolites thereof in a sample are provided. A plurality of particles, each of which is coated with a capture agent having an affinity for estradiol, is combined with the sample to form a plurality of estradiol-particle complexes. The system also includes a transport arrangement for transporting the sample and/or particles to the sensor surface, and optionally a magnetic field inducing structure constructed and arranged to establish a magnetic field at and adjacent to the sensor surface. The resonant sensor produces a signal corresponding to an amount of estradiol-particle complexes that are bound to the sensor surface.
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Citations
15 Claims
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1. A method for detecting estradiol in a sample, the method comprising the steps of:
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a) introducing a fluid comprising a sample and a plurality of magnetic particles coated with a first capture agent capable of binding estradiol into a fluid chamber, wherein at least one surface of the fluid chamber comprises a flexural plate wave device having a second capture agent capable of binding estradiol, wherein the flexural plate wave device has a membrane that is capable of vibrating, and wherein the second capture agent is bound to the membrane; b) applying a retractable source of magnetic flux positioned external to the fluid chamber close to the membrane to create a significant magnetic field gradient at a surface of the membrane to attract magnetic particles in the fluid toward the membrane; c) monitoring a first signal output by said flexural plate wave device, wherein the first signal output is monitored in the presence of a magnetic flux; d) after removing the retractable source of magnetic flux, flowing a solution through the fluid chamber to remove magnetic particles not bound to the membrane; e) monitoring a second signal output by said flexural plate wave device, wherein the second signal output is monitored in the absence of the magnetic flux; and f) comparing the first and the second signal output to a control signal, thereby detecting estradiol in the sample.
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2. The method of claim 1, wherein an increased level of estradiol is detected compared to a reference, which indicates that an individual from whom the sample is obtained has an increased likelihood of developing breast cancer.
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3. The method of claim 1, wherein prior to being introduced into the fluid chamber, said plurality of particles has been exposed to the sample.
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4. The method of claim 1, wherein the sample has been introduced into the fluid chamber prior to introducing the plurality of particles.
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5. The method of claim 1, wherein a concentration of about 10 pg/ml is detected.
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6. The method of claim 1, wherein the estradiol is selected from the group consisting of free estradiol, sex hormone-binding globulin (SHBG)-bound estradiol, estrone-3-glucuronide, and estradiol-3-glucuronide.
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7. The method of claim 1, wherein the sample is selected from the group consisting of blood, serum, plasma, urine, saliva, and biopsy material.
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8. The method of claim 1, wherein the control signal is a standard curve.
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9. The method of claim 1, wherein the control signal is obtained without the sample.
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10. The method of claim 1, wherein said second capture agent capable of binding the estradiol is indirectly bound to said surface.
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11. The method of claim 10, wherein said surface is coated with a first member of a binding pair, and the second capture agent is bound to a second member of the binding pair.
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12. The method of claim 11, wherein the first member of the binding pair is biotin.
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13. The method of claim 1, wherein the first or second capture agent is an antibody.
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14. The method of claim 11, wherein the second member of the binding pair is a derivative of avidin.
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15. The method of claim 1, wherein a fluid flows across the membrane during one or more of steps a), b), c) and e).
Specification