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WO2007074348A2 - Biocapteurs d'analytes multiples bases sur des transducteurs optoelectroniques - Google Patents

Biocapteurs d'analytes multiples bases sur des transducteurs optoelectroniques Download PDF

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Publication number
WO2007074348A2
WO2007074348A2 PCT/GR2006/000069 GR2006000069W WO2007074348A2 WO 2007074348 A2 WO2007074348 A2 WO 2007074348A2 GR 2006000069 W GR2006000069 W GR 2006000069W WO 2007074348 A2 WO2007074348 A2 WO 2007074348A2
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WO
WIPO (PCT)
Prior art keywords
optical
light
waveguide
biomolecules
biosensor
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Ceased
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PCT/GR2006/000069
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English (en)
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WO2007074348A3 (fr
Inventor
Sotirios Kakabakos
Konstantinos Misiakos
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National Center for Scientific Research Demokritos
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National Center for Scientific Research Demokritos
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Publication of WO2007074348A3 publication Critical patent/WO2007074348A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • G01N21/552—Attenuated total reflection
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/7703—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/7703—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides
    • G01N2021/7706—Reagent provision
    • G01N2021/7709—Distributed reagent, e.g. over length of guide
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7756—Sensor type
    • G01N2021/7763—Sample through flow
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N2021/7769—Measurement method of reaction-produced change in sensor
    • G01N2021/7776—Index

Definitions

  • the invention relates to the field of optical multianalyte biosensors with multiplexed output signal.
  • analytical microsystems be fabricated monolithically. So far, analytical microsystems are assembled by the hybrid integration of a components set. However, hybrid integration is costly, it complicates packaging and besides, it becomes impractical in multi-functional multi-target detection microsystems. This represents a problem especially for optical Microsystems, where precise alignment of the light sources to the transducer and detector subsystem is of critical importance and determines performance.
  • the present invention presents a solution to this need through the simultaneous fabrication of a large number of microsystem chips by the processing of large diameter silicon wafers.
  • the present invention addresses some key issues in order for such bioanalytical Microsystems to be able to compete even more effectively with traditional analytical techniques. Such key issues are
  • the invention advantageously employs optical detection to take advantage of the galvanic isolation of the transducer from the excitation and detection electronics.
  • Such a monolithic optical bioanalytical device is disclosed in this patent, which device advantageously uses the benefits of optical detection in a self-aligned monolithic silicon optical micro structure featuring single node readout for a large number of optocouplers.
  • the invention further enables label-free detection based on the use of noble metal nanoparticles or periodic-photonic formations on the waveguide.
  • the invention embodies two major innovations. More specifically, the monolithic optoelectronic transducer presented in WO03046527 (Al) and in K. Misiakos et al. Anal. Chem. 76, 1366-1373, 2004, consists of an array of silicon optocouplers made of silicon Light Emitting Diodes (LEDs), silicon nitride waveguides and silicon p-n junction detectors. The silicon LEDs are avalanche diodes biased beyond their breakdown voltage.
  • LEDs Light Emitting Diodes
  • the silicon nitride fibers are self aligned with respect to the silicon nitride waveguides by employing a silicon dioxide spacer and implantation of the avalanche diode emitter through the silicon nitride layer.
  • the silicon nitride waveguide optically connects the light emitter to the detector so that a monolithic silicon-based optocoupler is built.
  • the waveguide is functionalized by immobilizing capture biomolecules that later recognize the analyte biomolecules in the sample and bind to them.
  • photon absorbing labels like chromophore groups or metal nanoparticles are employed in order to follow the binding reaction of the biomolecules immobilized on the waveguide with the analyte in the sample.
  • the device described in the above mentioned references employs a number of independent optocouplers where each optocoupler has its own detector.
  • a single node output for the readout of all optocouplers of the device greatly facilitates the data collection process and greatly simplifies control electronics since a single readout electronics chain is needed and the multiplexing is enabled by selecting one emitter at a time.
  • Label-free detection is enabled in real time of analytes by exploiting the photon extinction shifts in spectral content and magnitude when biomolecules bind on noble nanoparticles immobilized on the waveguide surface.
  • label-free detection can be achieved by introducing on the waveguide surface patterns of another dielectric that make the optocouplers coupling efficiency sensitive to the effective refractive index of the superstrate.
  • the invention provides a monolithic optical biosensor for sensing biomolecules, comprising: a plurality of light-emitting elements; a plurality of optical waveguides, each waveguide optically coupled to a respective one of the plurality of light-emitting elements; and one single optical detector optically coupled to each of the plurality of optical waveguides, wherein the optical biosensor is adapted to detect an analyte by binding of the analyte on a surface of one or more of the waveguides and thereby changing the optical coupling between the optical detector and one or more of the light emitting elements.
  • the invention provides a monolithic optical biosensor for sensing biomolecules, comprising: a light emitting element; an optical waveguide optically coupled to the light emitting element; and an optical detector optically coupled to the optical waveguide, the optical waveguide having a plurality of surface features, the surface features being adapted to effect a change in the optical coupling between the light emitting element and the optical detector when biomolecules to be sensed are provided over the waveguide.
  • the invention provides a method of sensing biomolecules, comprising: providing a monolithic optical biosensor for sensing biomolecules, comprising: a plurality of light-emitting elements; a plurality of optical waveguides, each waveguide optically coupled to a respective one of the plurality of light-emitting elements; and one single optical detector optically coupled to the plurality of optical waveguides, wherein the optical biosensor device is adapted to detect an analyte by binding of the analyte on a surface of one or more of the waveguides and thereby changing the optical coupling between the optical detector and one or more of the light emitting elements; applying a solution containing the analyte to be sensed to the surface of one or more of the waveguides; and detecting a change of optical coupling between one or more of the light- emitting elements and the optical detector by measuring a change in detected light from one or more of the light-emitting elements by the optical detector.
  • the invention provides a method of sensing biomolecules, comprising: providing a monolithic optical biosensor for sensing biomolecules, comprising: a light emitting element; an optical waveguide optically coupled to the light emitting element; and an optical detector optically coupled to the optical waveguide, the optical waveguide having a plurality of surface features, the surface features being adapted to effect a change in optical coupling between the light- emitting element and the optical detector when biomolecules to be sensed are provided over the waveguide; applying biomolecules to be sensed over the surface of the waveguide; and detecting a change of optical coupling between the light-emitting element and the optical detector by measuring a change in detected light from the light-emitting element by the optical detector.
  • the invention provides a method of sensing biomolecules, comprising: providing a monolithic optical biosensor for sensing biomolecules, comprising: a plurality of light -emitting elements; a plurality of optical waveguides, each waveguide optically coupled to a respective one of the plurality of light emitting elements; and one single optical detector optically coupled to the plurality of optical waveguides, the optical waveguides having a plurality of surface features, the surface features being adapted to effect a change in optical coupling between the light- emitting elements and the single optical detector when biomolecules to be sensed are provided over the waveguides; applying biomolecules to be sensed over the surface of the waveguides; and detecting a change of optical coupling between the light-emitting elements and the optical detector by measuring a change in detected light one at a time from the light-emitting elements by the optical detector.
  • a multianalyte biosensor based on an array of monolithically integrated optoelectronic silicon transducers comprising avalanche diode light sources, silicon nitride optical fibers with multiplexed output signal on a single detector which is fabricated following silicon integrated circuit methods and can detect simultaneously multiple analytes in the same sample through either: the use of appropriately labeled biomolecules or surface nano-engineering that enables label-free detection of binding reactions in real time due to the change of the optical coupling between the integrated light sources and the integrated single node detector that is caused by the binding of the analytes on their respective recognition-molecules which have been previously immobilized on the plain (or nano-engineered) surface of the optical fibers that connect the light sources with the detector.
  • the invention presents a single node output for the readout of all optocouplers.
  • the invention may also enable label-free detection in real time of analytes by exploiting the photon extinction shifts in spectral content and magnitude when biomolecules bind on noble nanoparticles immobilized on the waveguide surface.
  • label-free detection can be achieved by introducing on the waveguide surface patterns of another dielectric that make the optocouplers' coupling efficiency sensitive to the effective refractive index of the superstrate.
  • An important element of one aspect of the present invention is surface nanoengineering of the fiber surface with isolated gold nanoparticles that permits label-free detection of bioreactions in real time.
  • Figure Ia shows a top view schematic of a set of monolithic optocouplers sharing the same detector node 13 ;
  • Figure Ib shows the cross section of the single p/n detector 13 where a number of independent waveguides 12 converge
  • Figure 2 presents a schematic diagram of one of the optocouplers after modification of the waveguide 23 with the metal nanoparticles 24
  • Figure 3 shows a schematic diagram showing surface patterns 34 on the waveguide 23 that make the optocouplers coupling efficiency sensitive to the effective refractive index of the superstrate;
  • Figure 4 presents the signal changes obtained from an optical waveguide with surface modification as in Fig. 2, during adsorption of anti-mouse IgG antibody on to gold nanoparticles, immunoreaction with mouse IgG and irnmunoreaction of the immobilized mouse IgG with anti-mouse IgG antibody in solution;
  • Figure 5 presents signal changes obtained from an optical waveguide with surface modification as in Fig. 3 during flow of distilled water followed by phosphate buffered saline reflecting changes in the effective refractive index of the waveguide superstrate medium.
  • the number of optocouplers shown in Fig. Ia is only indicative and can be expanded to any number with limits imposed only by the size of the chip, lithography and optocoupler layout. Such a configuration greatly facilitates the data collection process and greatly simplifies control electronics since a single readout electronics chain is needed and the multiplexing is enabled by selecting one emitter at a time.
  • the light emitting elements 11 are preferably adapted to be independently selected, so that the output from the detector 13 may be indicative of a selected one or more of the waveguides 12.
  • the light emitting elements 11 of the present invention are preferably avalanche diode light sources, although other light sources may also be suitable.
  • Fig Ib only the waveguides 12 coming from left, right and from top are shown.
  • the different waveguides deliver their photons independently to the detector p/n junction mainly at the abrupt breaking point of the waveguide at the edge of the p/n junction.
  • a large number of optocouplers can be measured with a single detector 13.
  • each waveguide 12 has a light emitting element 11 optically coupled at a first end and an optical detector 13 optically coupled at a second end.
  • light emitting elements and optical detectors are optically coupled at intermediate positions along the length of a waveguide.
  • light emitted by the light emitting elements of the present invention may be of a wavelength outside the normal visible spectrum, for example in the infra-red or ultraviolet regions.
  • FIG 2 is presented a schematic of one of the optocouplers after modification of the waveguide 23 with noble metal nanoparticles 24.
  • the nanoparticles 24 are functionalized with capture biomolecules that bind counterpart biomolecules and thereby change the nanoparticle photon extinction annihilation characteristics in spectral content and magnitude. This change is measured by monitoring the detector photocurrent changes with biomolecular binding. Binding events occurring between biomolecules close to the surface of a noble metal nanoparticle increase the refractive index of the nanoparticle' s immediate environment and cause a red shift of the homogeneous nanoparticle plasmon resonance [D. Eck, CA. Helm, Langmuir 17, 957-960,2001 ;A. fostl, K.
  • the transducers are first cleaned and hydrophylized in oxygen plasma. Then the transducers are immersed in solution of aminosilane or other silane that exposes reactive chemical groups for coupling of biomolecules or make the surface appropriate for adsorption of biomolecules. After that biotinylated bovine serum albumin or other biotinylated protein molecule is adsorbed onto the surface of the fiber. Then, the free binding sites on the surface of the fiber are covered with a solution of bovine serum albumin.
  • an appropriate microfluidic module adapted to transfer solutions over the fibers while insulating the electrical contact pads is applied on top of the transducer.
  • a streptavidin colloidal gold conjugate is pumped and the output signal current of the transducer is monitored.
  • the transucer is washed extensively with doubly distilled water and then oxidized in oxygen plasma. This way single plain gold colloidal nanoparticles are created on the fiber surface.
  • the so prepared gold particles can couple proteins or thiolated DNA oligonucleotides through self assemply of the thiol groups on the gold surface whereas the oxidized silicon nitride or silicon oxide layer surface do not adsorb biomolecules for a certain period of time.
  • Other deposition methods such as sputtering or adsorption of colloidal gold particles from solution could be used for surface nanoengineering of the fibers preferably provided that a decrease of the transducer photocurrent caused by the immobilized nanoparticles is in the range of 70-80% of its initial value (i.e. prior to nanoparticle application).
  • Another important innovation of the present invention that will permit label-free detection is achieved by introducing on the waveguide surface patterns of another dielectric material that makes the optocouplers' coupling efficiency sensitive to the effective refractive index of the superstrate.
  • FIG 3 a schematic drawing shows surface patterns 34 on the waveguide 23 that make the optocouplers coupling efficiency sensitive to the effective refractive index of the superstrate.
  • Fabrication of the grating involves a chemical vapor deposition step of SiO 2 over the Silicon Nitride waveguide followed by lithography and etching of the expose regions. Since these periods are quite long, the etching can be performed through wet chemistry, which has the additional advantage of good selectivity between the silicon dioxide (high etching rate) and the silicon nitride (much lower etching rate).
  • the waveguided modes enter from regions with no cladding layer 33 to regions with a cladding layer 33.
  • the detector 22 Each time the photons cross a different region reflection losses are experienced so that the photocurrent at the detector 22 is reduced with respect to the case of a uniform cladding layer.
  • the refractive index of the superstrate affects the reflection coefficient and the overall transmission constant. Therefore, measuring the detector photocurrent changes with different superstrate media or overlayers provides a measure for the effective refractive index of the overlaying medium. Therefore if biomolecules bind on the exposed waveguide 34 either directly or indirectly through another immobilized biomolecule, the detector 22 should sense a different photocurrent since the effective refractive index in the vicinity of the exposed waveguide surface should increase.
  • An additional reason for the long period grating is to contain the overall transmission coefficient into a realistic range so that enough light arrives at the detector.
  • a waveguide of the invention is adapted to detect biomolecules provided over the waveguide.
  • biomolecules are provided by passing an analyte solution over the waveguide such that the biomolecules therein are able to bind or otherwise associate with the waveguide surface or with surface features provided thereon and to consequently influence the optical path between a light emitting element and a detector coupled to the waveguide.
  • the optical path may be influenced by refraction, diffraction, absorption or other changes in light interactions resulting from the presence of biomolecules proximate the waveguide which thereby affect the level of light received by the optical detector.
  • a microfiuidic device On a wafer with a device in the form as illustrated in figure 1, a microfiuidic device is applied that has been appropriately designed in order to allow supply of reagents solution in all fibers of a single device simultaneously while it insulates the contact pads.
  • This microfiuidic module all the reagents solutions were run and the detector output signal recorded with the sampling rate of 1 sample/second ( Figure 2).
  • a 50 mM phosphate buffered saline, pH 7.4 (PBS buffer) was pumped onto the surface in order to establish a baseline for 3 min at a rate of 20 d/min (arrow 1).
  • a microfluidic device described in Example 1 is applied.
  • distilled water is pumped for 80 sec at a rate of 20 ⁇ Vmin ( Figure 5, arrow 51) over the waveguides of the array of the transducers and the output signal of the detector is recorded with a sampling rate of 1 sample/second.
  • a 50 mM phosphate buffered saline, pH 7.4 was pumped over the waveguides (arrow 52).
  • a change of the output signal is clearly demonstrated, indicating that the proposed surface modification of the waveguides is sensitive to refractive index changes.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Plasma & Fusion (AREA)
  • Engineering & Computer Science (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

L'invention concerne des biocapteurs optiques monolithiques permettant de détecter des biomolécules. Selon un aspect, les biocapteurs comprennent une pluralité d'éléments électroluminescents et des guides d'ondes optiques, chaque guide d'ondes étant optiquement couplé à un élément électroluminescent; ainsi qu'un détecteur optique, le biocapteur optique étant conçu pour détecter un analyte par liaison de l'analyte sur une surface d'au moins des guides d'ondes et modifiant ainsi le couplage optique entre le détecteur optique et au moins un des éléments électroluminescents. Selon un autre aspect, un biocapteur comprend un élément électroluminescent, un guide d'ondes optique optiquement couplé à l'élément électroluminescent et à un détecteur optique, le guide d'ondes optique présentant une pluralité d'éléments de surface conçus pour entraîner une modification de couplage optique entre l'élément électroluminescent et le détecteur optique lorsque des biomolécules à détecter sont présentes sur le guide d'ondes.
PCT/GR2006/000069 2005-12-27 2006-12-27 Biocapteurs d'analytes multiples bases sur des transducteurs optoelectroniques Ceased WO2007074348A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GR20050100623A GR20050100623A (el) 2005-12-27 2005-12-27 Βιοαισθητηρες ταυτοχρονης ανιχνευσης πολλαπλων αναλυτων που βασιζονται σε μονολιθικα ολοκληρωμενουσοπτοηλεκτρονικους μεταλλακτες σηματος
GR20050100623 2005-12-27

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WO2007074348A2 true WO2007074348A2 (fr) 2007-07-05
WO2007074348A3 WO2007074348A3 (fr) 2007-10-11

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4407299A1 (fr) * 2023-01-26 2024-07-31 Infineon Technologies AG Capteur de fluide pour détecter un fluide cible

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US5253037A (en) * 1992-08-04 1993-10-12 Fci-Fiberchem, Inc. Optimal length for refractive index sensors
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US7595189B2 (en) * 1999-01-08 2009-09-29 Applied Biosystems, Llc Integrated optics fiber array
EP1190236A1 (fr) * 1999-06-05 2002-03-27 Zeptosens AG Plate-forme de capteurs et procede permettant de determiner plusieurs substances a analyser
US6137117A (en) * 1999-06-21 2000-10-24 The United States Of America As Represented By The Secretary Of The Navy Integrating multi-waveguide sensor
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GB0117230D0 (en) * 2001-07-14 2001-09-05 Marconi Applied Technologies Detecting analytes
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GR1004178B (el) * 2001-11-29 2003-03-05 "����������" Ολοκληρωμενος οπτοηλεκτρονικος βιοαισθητηρας πυριτιου για ανιχνευση βιομοριων επισημασμενων με χρωμοφορες ομαδες ή νανοσωματιδια
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4407299A1 (fr) * 2023-01-26 2024-07-31 Infineon Technologies AG Capteur de fluide pour détecter un fluide cible

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GR20050100623A (el) 2007-07-23
GR1005589B (el) 2007-07-18
WO2007074348A3 (fr) 2007-10-11

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