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A method for sensing a chemical

A technology for detecting samples and samples, applied in the field of sensing chemicals, can solve the problem of not direct measurement of reporter antigen replacement

Inactive Publication Date: 2011-02-23
VIVACTA LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, reporter antigen displacement measurements are too indirect

Method used

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  • A method for sensing a chemical
  • A method for sensing a chemical
  • A method for sensing a chemical

Examples

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example 1

[0053] Preparation of Active Piezoelectric / Hot Film Biosensors

[0054] The ITO-coated polarized polyvinylidene fluoride (PVDF) bimorph piezoelectric film used as the sensing device in the following example was dip-coated in a polystyrene solution (1% toluene) in a low-humidity environment to A polystyrene layer is provided over the indium tin oxide. The bimorph piezoelectric film was then incubated overnight at room temperature to dissolve in a polystreptavidin solution (200 μg / mL in PBS, a pH containing 2.7 mmol / L KCl, 137 mmol / L NaCl, and 0.05% Tween 7.510mmol / L phosphate buffer). The preparation of polystreptavidin is described by Tischer et al. (US 5,061,640).

[0055] To prepare "capture" surfaces, polystreptavidin surfaces were incubated with biotin-labeled anti-testosterone to provide an antibody-coated surface (C1), or with biotin-labeled testosterone to provide an antigen-coated surface (C2 ). For C1, 10 ug / mL of biotin-labeled anti-testosterone (HyTest Ltd, Turk...

example 2

[0057] Preparation of carbon-labeled reporter conjugates

[0058] Carbon-labeled reporter conjugates were prepared essentially as described by Van Doom et al. (US 5,641,689). To prepare antibody-coated reporter conjugates (R1), incubate 1 mL of Special Black-4 RCC nominal 150nm with 200 μg / mL polystreptavidin solution in 5 mmol / L phosphate buffer pH 6.2 at room temperature with shaking Carbon microparticles (Degussa, Essen, Germany) were left overnight to give a streptavidin-coated surface (Al). The resulting carbon composites are washed (by centrifugation, pelleting and resuspension). 1 ml of this suspension of streptavidin-coated carbon particles was then incubated with 10 ug / mL of biotin-labeled anti-testosterone (HyTest Ltd, Turku, Finland, Cat # 2T2-biotin, or Accurate Chemical Co, Westbury, New York, USA, Cat # BHS113). The resulting carbon conjugates were washed 3 times with 0.05 mol / L borate buffer at pH 8.5 (by centrifugation, pelleting and resuspension) and stored...

example 3

[0060] Preparation of gold-labeled reporter conjugates

[0061] Gold-labeled reporter conjugates were prepared essentially as described by Frens G. Nature 1973, 241, 20-22 or Roth J. The colloidal gold marker system targets the field of light and electron microscopy cytochemistry. See Bullock GR, Petrusz P, eds. Techniques in immunocytochemistry, VoI 2. New York, NY, Academic Press, 1983, 216-284. To prepare antibody-coated reporter conjugates (R3), 1 mL of monodisperse nominal 150 nm gold particles was incubated with 200 μg / mL polystreptavidin solution in 5 mmol / L phosphate buffer pH 6.2 at room temperature with shaking (BBI International, Cardiff, UK) overnight to obtain a streptavidin-coated surface (A2). The resulting gold conjugates are washed (by centrifugation, pelleting and resuspension). 1 ml of this suspension of streptavidin-coated gold particles was then incubated with 10 ug / mL of biotin-labeled anti-testosterone (HyTest Ltd, Turku, Finland, Cat # 2T2-biotin, or...

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Abstract

The present invention relates to a method for detecting an analyte (10) in a sample, comprising the steps of: providing a transducer comprising a pyroelectric or piezoelectric element and electrodes which is capable of transducing a change in energy to an electrical signal, a first reagent immobilised on the transducer, and a second reagent (11) releasably bound to the first reagent and having a label attached thereto which is capable of absorbing electromagnetic radiation to generate energy by non-radiative decay, wherein either the first or second reagent has a binding site which allows binding to the other and which is capable of preferentially binding to the analyte or a derivative of the analyte; exposing the transducer to the sample thereby allowing the analyte or a derivative of the analyte to bind to the binding site and displace the second reagent; irradiating the sample with electromagnetic radiation; transducing the energy generated into an electrical signal; and detecting the electrical signal. The invention also provides a device for carrying out the method.

Description

technical field [0001] The present invention relates to methods for sensing chemicals, and more particularly to immunoassays utilizing chemical sensing devices comprising piezoelectric / pyroelectric transducers. Background technique [0002] Immunoassays are tests that measure the presence, or more generally, the concentration of an analyte in a biological fluid. It usually involves the specific binding of an antigen to an antibody. Antibodies can be polyclonal or monoclonal, and monoclonal antibodies have many benefits, including production reproducibility and inhibition of binding to one epitope of the analyte. To provide a quantifiable measure of the analyte concentration, the response is compared to a standard sample of known concentration. The concentration of antibody or antigen can be determined by a variety of methods, although one of the most common methods is to label the antigen or antibody and detect the presence of the label. [0003] Immunoassays can be compe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/542G01N21/17G01N23/00
CPCG01N33/5438
Inventor T·J·N·卡特S·A·罗斯
Owner VIVACTA LTD
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