CN101646401B - Biocompatible scaffold for sensing proteins - Google Patents
Biocompatible scaffold for sensing proteins Download PDFInfo
- Publication number
- CN101646401B CN101646401B CN2007800497563A CN200780049756A CN101646401B CN 101646401 B CN101646401 B CN 101646401B CN 2007800497563 A CN2007800497563 A CN 2007800497563A CN 200780049756 A CN200780049756 A CN 200780049756A CN 101646401 B CN101646401 B CN 101646401B
- Authority
- CN
- China
- Prior art keywords
- nanofiber
- nano wire
- biological analyte
- matrix
- timbering material
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
This invention related to a biocompatible scaffold that can detect redox-active chemicals and biomolecules electrochemically. In one embodiment, the biocompatible scaffold includes a substrate, and a conductive layer of TiO2-containing nanowires or nano fibers formed on the substrate, wherein the conductive layer of TiO2-containing nanowires or nanofibers is formed with a pore structure, and wherein when the biocompatible scaffold is in contact with a biological analyte, one or more proteins of the biological analyte are immobilized on a surface of the conductive layer Of TiO2- containing nanowires or nanofibers so as to generate a measurable faradic current signal.
Description
The application submits to as the PCT international application with the name of Univ Arkansas Technology Dev F. (U.S. state-owned firm); Application is All Countries except that the U.S., and only applies for United States Patent (USP) on January 12nd, 2007 with the name of Z.Ryan Tian (United States citizen).
CROSS-REFERENCE TO RELATED PATENT
The application benefits from applicant Z.Ryan Tian and proposes on January 12nd, 2007, and application number is 11/653,189 (No. 189 application), " TiO by name
2Nanostructured, film and thin layer and preparation method thereof " patent application; and this patent application self is according to united states patent law 35U.S.C. § 119 (e); require applicant Z.Ryan Tian and Wenjun Dong to propose on January 12nd, 2006; U.S. Provisional Patent Application number is 60/758,492, " TiO by name
2Nano fibrous membrane and manufacturing approach thereof and application " and applicant Z.Ryan Tian and Wenjun Dong propose on March 23rd, 2006, application number is 60/785,649, " TiO by name
2Nanofiber, film and thin layer and manufacturing approach thereof and application " patent application, quote in this combination, with for referencial use.
The application also benefits from applicant Z.Ryan Tian and proposes on January 12nd, 2007, and application number is 11/653,192, " TiO by name
2Nanostructured, film and thin layer and application thereof " patent application; and this patent application self is according to united states patent law 35U.S.C. § 119 (e); require applicant Z.Ryan Tian and Wenjun Dong to propose on January 12nd, 2006; U.S. Provisional Patent Application number is 60/758,492, " TiO by name
2Nano fibrous membrane and manufacturing approach thereof and application " and applicant Z.Ryan Tian and Wenjun Dong propose on March 23rd, 2006, application number is 60/785,649, " TiO by name
2Nanofiber, film and thin layer and manufacturing approach thereof and application " patent application, and this application relates to unsettled No. 189 applications equally, quotes in this combination, with for referencial use.
Quote and discussed some lists of references in the description of the present invention, comprise patent, patent application and all kinds of publication.To these lists of references quote and discussion is only used for description of the present invention clearly is described, be not to admit that these lists of references are indication of the present invention " prior aries ".The document of all references and discussion is quoted with same degree set at this in description, just as these documents combine reference separately.Form with [n] shows that it is the documents that a n piece of writing is quoted in the list of references tabulation hereinafter.For example, [15] are the 15th piece of list of references of quoting in list of documents, just, and Dong, W.; Cogbill, A.; Zhang, T.; Ghosh, S.; Tian, R.Z.J.Phys.Chem.2006,16819.
Technical field
The present invention relates generally to biosensor, more particularly, relate to a kind of bio-compatible nano wire (nanowire) timbering material, this timbering material is applicable to one or more albumen in the detection of biological analyte and can be used for supporting the cell growth.
Background technology
In the hole of the micron size of biologic bracket material, the activity of some albumen (as growth hormone GH) is vital [1] for tissue regeneration, and these tissues comprise that neuron cluster knits [2] and osseous tissue [3,4].Yet the protein active in monitoring and the detection of biological timbering material still faces the challenge.Develop a kind of electrochemistry albumen pick off that can accept the simple economy of this challenge, between albumen and the Solid Electrode Surface in Voltammetric Measurement fast electron transfer be essential condition.Yet, based on polypeptide-and/or the common electrical conductivity of biologic bracket material of polymer is lower.
Therefore redox protein like hemoprotein, carries electro-chemical activity, and can directly exchange electronics with the generation electrochemical signals with working electrode.This make the mediation method that need not (as through the enzyme mediated responses) but just Direct Electrochemistry detect with quantitatively.
Usually, need to rely on the lab analysis technology to come quantitative redox protein, as through the HPLC on the detector that is connected to a series of porous electrodes that are included in different potentials.Though this system is very accurate, tests required space and expense and make it be not suitable for large-scale pattern detection or domestic consumer's use.In addition, biological sample generally it is become can be by the form of HPLC analysis.
Like this; Though it is accurate inadequately to be used for the bio-sensing device of quantitative redox protein; Develop a kind ofly be easy to carry, disposable and can analyze the bio-sensing device of simple pretreated biological sample; Thereby in multichannel in real time and need not laboratory and detect and prior art is improved aspect proteic, caused people's very big interest.
Therefore, up to now, above-mentioned defective and deficiency still exist in the art, and need to solve.
Summary of the invention
In one aspect, the present invention relates to a kind of bio-compatible timbering material.
In one embodiment, said bio-compatible timbering material comprise matrix, on said matrix, form contain TiO
2Nano wire or nanofiber conductive layer, the said TiO that contains
2Nano wire or nanofiber conductive layer form with pore structure, and when said bio-compatible timbering material contacts with biological analyte, one or more albumen of said biological analyte will be fixed on the said TiO of containing
2The surface of nano wire or nanofiber conductive layer on, and then generate measurable sensor current signal.
The said TiO that contains
2Nano wire or nanofiber mainly be TiO
2-B phase or metatitanic acid salt face, and the representative diameter scope that has is about 20-150nm.
In one embodiment, the thickness range of said pore structure is about 1-50 μ m.Said nano wire or nanofibrous structures can have layering titanate (Na
2Ti
3O
7) structure, this structure has the cation of liquidating (counter-cations) (Na between its interlayer
+).This matrix is made up of tin indium oxide (ITO) or metal.
In one embodiment, said albumen comprises the albumen with redox active center.Said redox active hemoprotein comprises cytochrome c.
Said matrix can be conductor or quasiconductor.In one embodiment, said matrix is made up of tin indium oxide (ITO).
In another aspect, the present invention relates to the bio-compatible timbering material.In one embodiment, this bio-compatible timbering material has matrix, and nano wire that on this matrix, forms or nanofiber layer.
The nano wire or the nanofiber layer that form have pore structure.This nano wire or nanofiber comprise oxidiferous nano wire or nanofiber.In a specific embodiment, said nano wire or nanofiber comprise and contain TiO
2Nano wire or nanofiber.
In another aspect, the present invention relates to be used for one or more proteic biosensors of detection of biological analyte always.In one embodiment; This biosensor comprises the matrix with surface; Nano wire that on the surface of said matrix, forms or nanofiber layer have pore structure in this nano wire or the nanofiber layer of this formation, and said biosensor also comprises the detector of communicating by letter with this matrix; When this layer when nano wire or nanofiber contacts with one or more albumen of biological analyte, also can detect by this detector but will generate measuring-signal.
This biosensor further comprises the device that is used for the sweep speed scope of about 0.005-0.500V/s voltage being applied to biological analyte.In one embodiment, said detector comprises cyclic voltammetry scan appearance (cyclic voltammetry).When the layer that forms when nano wire or nanofiber contacts with one or more albumen in the biological analyte, but one or more electron transfers between biological analyte and the matrix will produce the measuring-signal of faradic current form.
In one embodiment, one or more albumen in the said biological analyte comprise the redox active hemoprotein.Said redox active hemoprotein comprises cytochrome c.This biological analyte comprises that the pH scope is about the buffer solution of 6.2-9.0, and cytochrome c and nano wire or nanofiber carry clean positive charge and net negative charge respectively like this.
Said nano wire or nanofiber are oxidiferous nano wire or nanofiber.In one embodiment, this nano wire or nanofiber are to contain TiO
2Nano wire or nanofiber.At this, this contains TiO
2Nano wire or nanofiber mainly be TiO
2-B phase or metatitanic acid salt face.In one embodiment, the representative diameter scope of said nano wire or nanotube is about 20-150nm.
In one embodiment, said matrix is made up of tin indium oxide (ITO).Said matrix can be conductor or quasiconductor.
In another aspect, the present invention relates to be used for one or more proteic methods of detection of biological analyte.In one embodiment; Said method comprises the step of the biosensor that the timbering material that has at least one electrode and be made up of nano wire or nanofiber is provided; Said biological analyte is imported to the step in the said timbering material; And the electron transfer between the surface of detection of biological analyte and said timbering material, with one or more proteic steps in the detection of biological analyte.In one embodiment, said detection step is accomplished by CV.
Said method further comprises the step that voltage is applied to biological analyte with the sweep speed scope of about 0.005-0.500V/s.
In one embodiment, said biological analyte comprises the redox active hemoprotein.Said redox active hemoprotein comprises cytochrome c.
This biological analyte comprises that the pH scope is about the buffer solution of 6.2-9.0, and cytochrome c and nano wire or nanofiber carry clean positive charge and net negative charge respectively like this.
Through following description, specific embodiment, combine accompanying drawing again, can more in depth understand various aspects of the present invention.Can carry out various changes or equivalence replacement and not exceed protection scope of the present invention these characteristics and embodiment.
Description of drawings
The accompanying drawing illustration one or more embodiment of the present invention, and principle of the present invention has been described with associated description.In the case of any possible, same Reference numeral is used for representing parts or the intimate parts that embodiment is identical in the accompanying drawing in each width of cloth accompanying drawing, wherein:
Fig. 1 schematically shows biosensor according to an embodiment of the invention;
Fig. 2 shows the phenogram according to embodiments of the invention about three days consuming time synthetic titanate nano wire (NW) timbering materials under about 180 ℃ temperature conditions respectively, (a) SEM of titanate NW timbering material figure, (b) TEM of titanate NW timbering material figure; (c) XRD figure of titanate NW timbering material; At this, this metatitanic acid salt face is 2 θ=9.8 ° (001), 11.2 ° (200); ° 24.4 (110) and 29.7 ° (003) (JCPDScard No.:47-0561); And (d) FT-IR spectrum, at this, curve 210 is the FT-IR spectrum that is fixed to the cytochrome c on titanate-NW timbering material; Wherein said cytochrome c has mixed KBr, and curve 220 is the FT-IR spectrum of the cytochrome c in the buffer solution of pH 6.8;
Fig. 3 shows sign (anode current, the i of the titanate NW timbering material shown in Fig. 2
Pa, and cathode current i
Pc), (a) (curve 330a is used for i to the CV curve of ito glass electrode in the buffer solution of pH6.8 of covering titanate NW
Pa, curve 330c is used for i
Pc) and the CV curve in the buffer solution of ito glass electrode that is fixed with the cytochrome c that 450 skins rub on the titanate NW at pH6.8 (curve 340a is used for i
Pa, curve 340c is used for i
Pc), be about 0.2V/s in this sweep speed; (b) the CV curve of the cytochrome c of various concentration in the buffer solution of pH6.8, wherein curve 351a is the i that 45 skins rub
Pa, curve 352a is the i that 122 skins rub
Pa, curve 353a is the i that 243 skins rub
Pa, curve 354a is the i that 347 skins rub
Pa, curve 355a is the i that 450 skins rub
Pa, and be about 0.2V/s in this sweep speed; (c) cytochrome c concentration and anode current i
PaLinear relationship; And (d) after cultivating 72 hours on the titanate NW timbering material, the SEM of the PC-12 cell of on titanate NW timbering material, growing figure;
Fig. 4 shows the sign of the titanate NW timbering material shown in Fig. 2, (a) after cultivating 72 hours on the titanate NW timbering material, and the SEM figure of the PC-12 cell of on titanate NW timbering material, growing; And (b) be fixed on cytochrome c on the titanate NW timbering material at the CV of different potential sweep speed and pH value curve;
Fig. 5 shows the sign of NW timbering material according to an embodiment of the invention;
Fig. 6 shows the sign of NW timbering material according to an embodiment of the invention.
The specific embodiment
In context of the present invention, use in the present invention in the specific context of these terms, the term that uses in this description has its general implication in the art.
Below or in other position of this description; Some is used to describe term of the present invention discusses; So that in description, and to the technical staff additional guidance is provided to the manufacturing of these apparatus and method with in using to apparatus and method of the present invention.For the ease of reading, some term has been carried out labelling, such as adopting italic and/or quotation marks.The use of these labellings does not influence the scope and the meaning of this term, and the scope of this term is identical with meaning, and in a part, no matter whether this term is labeled, and its implication is the same.Should be appreciated that same incident can have multiple saying.Therefore, can replacement term and synonym be used for any one or more terms that discuss in this place, no matter term is that prepare or mistake discussed herein, and it does not have any special implication.The synonym of some term also is provided.Other synon use is not got rid of in one or more synon narrations.The embodiment that uses in the present invention (example that comprises any term discussed herein) is only used for exemplaryly, is not limited to scope of the present invention and implication or any term of giving an example out.Same, the embodiment that the present invention is not limited to provide in the description.In addition, subhead can be used for helping the reader of description to read over whole description, yet the use of subhead does not influence scope of the present invention.
As use, " approximately " or " being similar to " refers generally in set-point or scope 20 percentage points, and in preferred 10 percentage points, the best is to be positioned in 5 percentage points. hereThe quantity that here provides is proximate, means that term " approximately " or " being similar to " can be derived if clearly do not explain.
As use, term " isopotential point " or its abbreviation " pI " are meant molecule or the surperficial pH of net charge that do not carry. hereIn order to have more sensitive pI, molecule (or surface) must be amphoteric, means it and must have acidity and basic group.Albumen and aminoacid are the common molecules that meets this requirement.
Albumen can separate according to its isopotential point through known isoelectrofocusing process.Be lower than the pH of pI, the clean positive charge of albumen portability.Be higher than pI, its portability net negative charge.But this means leakage of electricity swimming gel.The pH of running gel can be confirmed by the buffer solution that this gel uses.If the pH of this buffer solution is higher than proteic pI, this albumen will move to positive pole (negative charge will be attracted to positive pole).If the pH of this buffer solution is lower than proteic pI, this albumen will move to negative pole (positive charge will be attracted to negative pole).If albumen is to equal to run in the buffer solution of its pI electrophoretic at pH value, it will can not move.
As use, term " cyclic voltammetry " or its abbreviation " CV " are meant one type electrokinetic potential electrochemical measurement method. hereIn order to obtain cyclic voltammetric, can measure the variation that change in voltage and electric current in the solution produce along with change in voltage.This is a kind of particular type in the voltammetry of redox property of research chemistry and interfacial structure.
In cyclic voltammetry experiment, current potential is applied in the system, and measures faradic current response (faradic current is the electric current that redox reaction produces).Can measure this current-responsive (potential window) in the certain potentials scope, this voltage range arises from an initial value, and changes to a preset limit value with linear mode.This current potential (being commonly referred to switching electric potential (switching potential)), the counter-rotating of the direction of electric potential scanning, and the same potential window of reverse scan simultaneously.This means, and the element that for example forms in (forward) scanning oxidation for the first time will be reduced when scanning the second time (oppositely).This technology is usually used, because it provides the quick and simple method that is used for initial token redox active system.It rate information of electron transfer between electrode and the analyte also can be provided, and analyte is in the stability (for example, if it experiences any chemical reaction) of electrolytic oxidation state except the oxidation-reduction potential estimation also can be provided.
Integral viewpoint of the present invention
The bio-compatible timbering material is very important [1] to tissue regeneration, regenerates under the inducing of albumen such as growth hormone like osseous tissue [3,4].For proteic activity in the compatible timbering material of detection of biological, need a kind of electrochemistry albumen pick off of development, it has the support that can realize between albumen and sensor electrode that swift electron shifts.Yet, based on polypeptide-and/or the common electrical conductivity of biologic bracket material of polymer is lower.
Solid nano silk (NW) is considered to can be used for detecting single virus [5].Wherein, based on TiO
2The NW cost of manufacture low, bio-compatibility is high, chemistry and photochemical properties stable, and albuminous degeneration is little, [6-8] therefore is widely used.For example, hemoprotein can be fixed to TiO
2(NT) [9-14] on nano-particle and the nanotube.
In addition, the invention also discloses titanate-NW bio-compatible timbering material, it twines on the surface of tin indium oxide (ITO) matrix (electrode) each other.Titanate-timbering material is used as biosensor, or is the part of biosensor at least, is used to detect the electrochemical redox current potential that pre-fixes at the cytochrome c (cyt c) on titanate-NW surface.In addition, titanate-NW timbering material can be used for supporting the growth of pheochromocyte oncocyte (PC-12).
To combine embodiments of the invention and accompanying drawing 1-6 to describe.According to the object of the invention,, in one aspect of the invention, relate to the albumen that is used for automatic detection of biological analyte and the bio-compatible timbering material of support and guidance stem cell growth like concrete and general description at this.
Said bio-compatible timbering material comprise matrix, on said matrix, form contain TiO
2Nano wire or nanofiber conductive layer.This matrix can be conductor or quasiconductor.In one embodiment, said matrix is made up of tin indium oxide (ITO).The said TiO that contains
2Nano wire or nanofiber conductive layer form with pore structure, the thickness range of this pore structure is 1-50 μ m.This pore structure can have layering titanate (Na
2Ti
3O
7) structure, this structure has the cation of liquidating (counter-cations) (Na between its interlayer
+).The said TiO that contains
2NW mainly be TiO
2-B phase or metatitanic acid salt face, and the representative diameter scope that has is about 20-150nm.
Other NW that contains interlayer cation also can be used for realizing the present invention.
When said bio-compatible timbering material contacts with biological analyte, one or more albumen of said biological analyte will be fixed on the said TiO of containing
2The surface of nano wire or nanofiber conductive layer on, this can biological analyte with contain TiO
2Nano wire or nanofiber conductive layer surface between electron transfer will appear.This electron transfer will generate measurable sensor current signal.
This bio-compatible timbering material can be used for the redox active hemoprotein in the detection of biological analyte.Cytochrome c (cyt c) is the hemoprotein of redox active, and it has 104 aminoacid, and is positioned at mitochondrial subintimal space as the electron carrier in the electronics conveyer chain.It is a soluble protein, and can but can not combine oxygen atom through oxidated and reduction.Be about in the buffer solution of 6.2-9.0 in the pH scope, cytochrome c carries clean positive charge, and nano wire or nanofiber carry net negative charge.When being fixed on, cytochrome c contains TiO
2NW timbering material surperficial the time, but one or more electron transfers between cytochrome c and the NW timbering material will produce measuring-signal with faradic form.This can be detected by cyclic voltammetry (CV).Also can adopt other albumen to realize the present invention.
With reference to Fig. 1, show biosensor 100 according to an embodiment of the invention, one or more albumen that this biosensor 100 uses NW timbering material 120 to come in the detection of biological analyte.
The NW timbering material can be used for sustenticular cell, like the growth of pheochromocyte oncocyte (PC-12).In one embodiment.This NW timbering material part has at least covered the large number of biological molecule, comprises growth hormone.
Another aspect of the present invention relates to and is used for one or more proteic methods of detection of biological analyte.In one embodiment, said method comprises the following steps, biosensor at first is provided, the timbering material that this biosensor has electrode and on this electrode, formed by NW.Then said biological analyte is imported in the said timbering material.Then adopt the electron transfer between CV detection of biological analyte and the said matrix surface, this CV is used in the detection of biological analyte whether albumen and/or its amount being arranged.
Do explanation further in the face of of the present invention these with others down.
Realization of the present invention and embodiment
Show typical method and its correlated results according to an embodiment of the invention below, the scope that these embodiment are not intended to limit the invention.It should be noted that the title used in an embodiment or subhead all are readers for ease, is not to limit scope of invention by any way.In addition; Mention and disclose some theories since then; Yet no matter these theoretical correctness are not the scopes of the present invention that limits by any way, can need not to consider any particular theory or arrangement of the exercise according to the description of the invention embodiment of the present invention fully like this.
Embodiment: be used for directly detecting automatically proteic bio-compatible timbering material
In this exemplary embodiments, the surface that titanate-NW is wrapped in the ITO matrix forms timbering material, and it can be used for monitoring the electrochemical redox current potential that pre-fixes at the cytochrome c on titanate-NW surface.This titanate-NW under about 240 ℃ temperature conditions about three days consuming time synthetic.
Fig. 2 a and 2b show the sem photograph (SEM) and the transmission electron microscope figure (TEM) of titanate-NW support respectively.The x-ray diffractogram of powder of titanate-NW support has been shown among Fig. 2 c.This X-ray diffraction is clear to have shown that the crystal structure of NW belongs to metatitanic acid salt face (Na just,
2Ti
3O
7), it is characterized by following lattice parameter: [2 θ=9.8 ° (001), 11.2 ° (200), 24.4 ° (110) and 29.7 ° (003) (JCPDScard No.:47-0561)].The TiO of (edge-shared) is shared at this edge
6-octahedron is the elementary cell [15] that forms this negative charge layering titanate structure, and this layering titanate structure has the sodium that the is arranged in inner space cation (Na that liquidates
+), will cause like this forming variable interlamellar spacing according to cationic size and degree of hydration.Therefore, this titanate NW timbering material has pore structure, shown in Fig. 2 a and 2b.After adopting deionized-distilled water (DDW near pH near 7) washing, this sodium cation that liquidates can be replaced by proton.
Usually, the impedance of air-dry NW-timbering material is higher than 10
6Ohm, this also is not suitable for electrochemistry and detects automatically.Yet; (OH) [4] will locate to produce the effect of wet electronics in water-NW interface [16] to the lip-deep hydroxyl of NW in the buffer solution of pH6.2-9; On the NW surface, being formed for low-energy path of quick ET, this will guarantee that conversion of signals can cross over the timbering material of whole bio-compatible and carry out in bio-sensing.In other words, in buffer solution, the surface of titanate NW can be negative charge and form Ti-OH [4].Interface between water and the oxidized metal can generate wet electronics [16], so the electric conductivity of titanate NW is improved.
Cytochrome c is oxidoreduction-active hemoprotein, has 104 aminoacid, and is positioned at mitochondrial subintimal space as the electron carrier in the electronics conveyer chain.In order to check after being attached to negative charged surface, whether cyt c no longer firm [17-19], can use fourier transform infrared spectrometry (FT-IR) with in the profiling protein to the responsive amino group of polypeptide structure.Usually, because along the stretching vibration of the C=O group of albumen main chain, said amino I signal is mainly at 1600-1700cm
-1Fig. 2 d shows FT-IR spectrum, and wherein curve 210 is the FT-IR spectrum that mixes cytochrome c-titanate NW of KBr, and curve 220 is the FT-IR spectrum of the cytochrome c in pH6.8 buffer solution.The peak value that this FT-IR spectrum is illustrated in the amino I of the last fixed cyt c of NW is 1648cm
-1, this and the same [20] that natural cyt c from the buffer solution of pH6.8 obtains show the residual secondary structure (retained secondary structure) that is fixed to the cyt c on titanate-NW timbering material.
The static that bonded cyt c mainly is attributed between negative charge NW surface and the positive charge cyt c surface on the titanate NW timbering material is mutual.At pH6.2-9.0, because its isopotential point is 6.2, the NW surface is negative charge [21], yet because its isopotential point is 10.0-10.5, cyt c surface is a positive charge.Like this, bear the NW surface the high captivation that aligns cyt c is shown.In addition; The shape of cyt c in water is almost spherical (a*b*c=1.5nm*1.7nm*1.7nm) [22]); Like this, cyt c can be easy to be fixed on the vacancy of timbering material (diameter＞500nm) is further to strengthen the stop of cyt c in the electrochemical redox process.
The NW timbering material provides ideal environment to carry out quick electron transfer reaction for cyt c.In this exemplary embodiments, can under different concentration, study the surface electrochemistry characteristic of the NW that is fixed with cytochrome c through CV, its 45 skins that indicate cyt c rub (or 10
-12Mole) detection limit.
Fig. 3 a shows respectively that (signal 330a is used for anode current i from being coated with NW
Pa, and signal 330c is used for cathode current i
Pc) and the CV signal of ito glass electrode in the buffer solution of potassium phosphate of pH=6.8 of cyt c-NW (signal 340a is used for anode current i
Pa, and signal 340c is used for cathode current i
Pc).Sweep speed is about 0.2V/s.Recorded a pair of apparent spike potential (E that has through cyt c-NW electrode
0) reversible and clear redox peak.This E
0Value (0.03V), from the average of anode spike potential value and the calculating of negative electrode spike potential value, approaching with the report of Stellwagen [23].Based on its structure and form, NW shows as conduction " nanometer cable " [16] so that electron transfer.
Can be used for the diagnostic electrode surface as the faradic measurement of sweep speed whether the redox reaction by DIFFUSION CONTROLLED is arranged.This anode and negative electrode peak value all with sweep speed linear correlation from 0.01V/s to 0.2V/s, this means that such electrode has typical thin layer electrochemistry characteristic [24].i
PcAnd i
PaRatio be about 2.0, and the interval of oxidoreduction peak value is about 58mV (data not shown goes out), this means that electrochemical process on the cytc-NW electrode surface can be will definitely contrary [24].
Shown in Fig. 3 b, in the buffer solution of pH6.8, this faradic current increases along with the concentration of the cytc in the timbering material is linear, is the i that 45 skins rub at this signal 351a
Pa, signal 352a is the i that 122 skins rub
Pa, signal 353a is the i that 243 skins rub
Pa, signal 354a is the i that 243 skins rub
Pa, signal 355a is the i that 347 skins rub
Pa, and be about 0.2V/s in this sweep speed.This is reversible and the CV signal can be between the μ M of 3.0 μ M and 30.0 clearly, or observes in the solution of 15 μ L between 45 to 450 skin massage.These concentration can be with the complete linear correlation of anode current (R=0.9887), shown in Fig. 3 c.
Its result shows that the ITO electrode that a kind of like this titanate NW modifies can be used as polynary automatic detection platform, is used to develop sane, sensitive, economic nano biological sensor.
In addition, this NW timbering material also can be used for the growth of support and guidance pheochromocyte oncocyte (PC-12), helps cytoactive at this.Fig. 3 d and 4a are the SEM figure that on the NW timbering material, cultivates about 72 hours PC-12 cell, show the PC-12 cell well attached to forming cell mass on the NW timbering material and on this material.These cells show as circle in shape, and keep its form, and this means between cell and the NW timbering material has favorable compatibility.The albumen that further means absorption attached to the PC-12 cell on the titanate NW timbering material is not lost any activity.
Fig. 4 b shows the albuminous cell pigment c that is fixed on the NW timbering material thin film at the CV of different potentials sweep speed and pH value signal.In the pH of 6.8-9.0 scope, but can obtain the duplicate detection lower limit of the cytochrome c that 45 skins rub, this has shown that the direct electron of the cytochrome c on the ITO electrode that titanate NW modifies shifts and has been strengthened greatly.The even current potential of the cytochrome c that NW is last depends on that pH changes.Its result makes for functionalization on ceramic nano silk surface and based on NW captivation-particular organisms nano-sensor new opinion is provided, and can produce the polynary automatic detection platform that is used to develop sane, sensitive, economic nano biological sensor.
Fig. 5 and 6 is the CV signals according to the NW timbering material of other embodiments of the invention.
In brief, the present invention among others, discloses a kind of titanate NW timbering material of bio-compatible, and a kind of biosensor based on titanate NW timbering material.This titanate NW timbering material has strengthened the electron transfer between ITO electrode and the cyt c.Should can be applicable in based on the constrained environment of titanate NW timbering material, in real time fixed redox active albumen directly detected based on the biosensor of titanate NW timbering material, and the Biochemical processes of monitoring in the growth course of stem cell.
Above-mentioned purpose of description to the preferred embodiments of the present invention is to explain for example and describe.These embodiment are not exhaustive, that is to say that the present invention does not receive the restriction of disclosed precise forms, under instruction of the present invention or from practice of the present invention, can obtain multiple modification and variation to these embodiment.
Selection and description to embodiment are for principle of the present invention and practical application thereof are made explanations, to enable those skilled in the art to utilizing the present invention in various embodiments and to serve as to cooperate specific use to carry out various modifications.Those skilled in the art know that in the alternative embodiment that does not break away from the principle and scope of the present invention.Therefore, scope of the present invention is claim of the present invention and be equal to and limit, rather than the description of front and exemplary embodiments described herein.
List of references
.Murugan，R.；Ramakrishna，S.Tissue?Eng.2006，12，435.
.Ellis-Behnke，R.G.；Liang，Y.；You，S.；Tay，D.K.；Zhang，S.；So，K.F.；Schneider，G.E.Proc.Natl.Acad.Sci.USA?2006，103，5054.
.Liao，S.；Cui，F.；Zhang，W.；Feng，Q.J.Biomed.Mater.Res.2004，69B(2)，158.
.Kokubo，T.；Kim，H.M.；Kawashita，M.Biomaterials?2003，24，2161.
.Patolsky，F.；Zheng，G.；Hayden，O.；Lakadamyali，M.；Zhuang，X.；Liber，C.M.Proc.Natl.Acad.Sci.USA?2004，101，14017.
.O’Regan，B.；
M.Nature?1991，353，737.
.Cosnier，S.；Gondran，C.；Senillou，A.；
M.；Vlachopoulos，N.Electroanalysis.1997，9，1387.
.Leng，Y.；Huang，N.；Yang，P.；Chen，J.；Sun，H.；Wang，J.；Wan，G..；Tian，X.；Fu，R.K.Y.；Wang，L.；Chu，P.K.Surf.Coat.Technol.2002，156(1-3)，295.
.McKenzie，K.J.；Marken，F.Langmuir?2003，19，4327.
.Topoglidis，E.；Cass，A.E.G.；O’Regan，B.；Durrant，J.R.J.electroanal.Chem.2001，517，20.
.Topoglidis，E.；Campbell，C.J.；Cass，A.E.G.；Durrant，J.R.Langmuir?2001，17，7899.
.Paddon，C.A.；Marken，F.Eletrochem.Commun.2004，6，1249.
.Zhou，H.；Gan，X.；Wang，J.；Zhu，X.；Li，G.Anal.Chem.2005，77，6102.
.Liu，A.；Wei，M.；Honma，I；Zhou，H.Anal.Chem.2005，77，8068.
.Dong，W.；Cogbill，A.；Zhang，T.；Ghosh，S.；Tian，R.Z.J.Phys.Chem.2006，16819.
.Onda，K.；Li，B.；Zhao，J.；Jordan，K，D.；Yang，J.；Petek，H.Science?2005，308，1154.
.Heimburg，T.；Marsh，D.Biophys.J.1993，65，2408.
.Pinheiro，T.J.T.；
G.A.；Watts，A.；Roder，H.Biochemistry.1997，36，13122.
.Sanghera，N.；Pinheiro，T.J.T.Protein?Sci.2000，9，1194.
.Paquet，M-J.；Laviolette，M.；Pézolet，M.；Auger，M.Biophys.J.2001，81，305.
.Shinohara，H.；
M.；Vlachopoulos，N.；Aizawa，M.Bioelectrochem.Bioenerg.1991，26，307.
.Baglioni，P.；Fratini，E.；Lonetti，B.；Chen，S，H.J.Phys：Condens.Matter.2004，16，s5003.
.Bard，A.J.；Faulkner，L.R.Electrochemical?Methods.Fundamentals?andApplications，2nd?ed；Wiley：New?York，2001.
.Barker，P.D.；Mauk，A.G.J.Am.Chem.Soc.1992，114，3619.
.Perroud，D.T.；Bokoch，P.M.；Zare，N.R.Proc.Natl.Acad.Sci.USA?2005，102，17570.
.Stellwagen，E.Nature?1978，275，73.
Claims (19)
1. bio-compatible timbering material comprises:
A. matrix; And
What b. on said matrix, form contains TiO
2Nano wire or nanofiber conductive layer, it is characterized in that the said TiO that contains
2Nano wire or nanofiber conductive layer form with pore structure, and when said bio-compatible timbering material contacts with biological analyte, one or more albumen of said biological analyte will be fixed on the said TiO of containing
2The surface of nano wire or nanofiber conductive layer on, and then generate measurable sensor current signal, the thickness range of said pore structure is 1-50 μ m, said pore structure comprises layering titanate structure Na
2Ti
3O
7, have the cation of liquidating Na between its interlayer
+, said matrix is made up of tin indium oxide.
2. bio-compatible timbering material according to claim 1 is characterized in that said albumen comprises the redox active hemoprotein.
3. bio-compatible timbering material according to claim 2 is characterized in that, said redox active hemoprotein comprises cytochrome c.
4. bio-compatible timbering material according to claim 1 is characterized in that, the said TiO that contains
2Nano wire or nanofiber mainly be TiO
2-B phase or metatitanic acid salt face.
5. bio-compatible timbering material according to claim 4 is characterized in that, the said TiO that contains
2The representative diameter scope that has of nano wire or nanofiber be 20-150nm.
6. be used for one or more proteic biosensors (100) of detection of biological analyte, comprise
A. the matrix (110) that has top layer (112);
B. go up nano wire or the nanofiber layer (120) that forms on the top layer (112) of said matrix (110); Said nano wire or nanofiber layer in this formation have pore structure; The thickness range of said pore structure is 1-50 μ m, and said pore structure comprises layering titanate structure Na
2Ti
3O
7, have the cation of liquidating Na between its interlayer
+, said matrix is made up of tin indium oxide; And
The detector (140) of c. communicating by letter with said matrix (110) when the layer (120) of nano wire or nanofiber formation contacts with one or more albumen of biological analyte, also can be detected by said detector (140) but will generate measuring-signal.
7. biosensor according to claim 6 is characterized in that, said one or more albumen comprise the redox active hemoprotein.
8. biosensor according to claim 7 is characterized in that, said redox active hemoprotein comprises cytochrome c.
9. biosensor according to claim 8 is characterized in that, said biological analyte comprises that the pH scope is the buffer solution of 6.2-9.0, and cytochrome c and nano wire or nanofiber carry clean positive charge and net negative charge respectively like this.
10. biosensor according to claim 6 is characterized in that, the representative diameter scope of said nano wire or nanofiber is 20-150nm.
11. biosensor according to claim 6 is characterized in that, said biosensor further comprises the device that is used for the sweep speed scope of 0.005-0.500V/s voltage being applied to biological analyte.
12. biosensor according to claim 11 is characterized in that, said detector comprises the cyclic voltammetry scan appearance.
13. biosensor according to claim 12; It is characterized in that; When the layer (120) that forms when nano wire or nanofiber contacts with one or more albumen in the biological analyte, but one or more electron transfers between biological analyte and the matrix will produce the measuring-signal of faradic current form.
14. one kind is used for one or more proteic methods of detection of biological analyte, it is characterized in that said method comprises the following steps:
A., biosensor is provided; Said biosensor has electrode and timbering material; Nano wire or nanofiber layer that said timbering material has matrix and forms on the surface of said matrix; Said nano wire or nanofiber layer in this formation have pore structure, and the thickness range of said pore structure is 1-50 μ m, and said pore structure comprises layering titanate structure Na
2Ti
3O
7, have the cation of liquidating Na between its interlayer
+, said matrix is made up of tin indium oxide;
B. said biological analyte is imported in the said timbering material; And
C. detect the electron transfer between said biological analyte and the said matrix surface, and then one or more albumen in the detection of biological analyte.
15., it is characterized in that said biological analyte comprises the redox active hemoprotein according to the said method of claim 14.
16., it is characterized in that said redox active hemoprotein comprises cytochrome c according to the said method of claim 15.
17., it is characterized in that said biological analyte comprises that the pH scope is the buffer solution of 6.2-9.0 according to the said method of claim 16, cytochrome c and nano wire or nanofiber carry clean positive charge and net negative charge respectively like this.
18. according to the said method of claim 14, it is characterized in that, further comprise the step that voltage is applied to biological analyte with the sweep speed scope of 0.005-0.500V/s.
19., it is characterized in that said measuring process is accomplished by cyclic voltammetry according to the said method of claim 18.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US11/653,189 | 2007-01-12 | ||
| US11/653,192 US7931812B2 (en) | 2006-01-12 | 2007-01-12 | TiO2 nanostructures, membranes and films, and applications of same |
| US11/653,189 US8883115B2 (en) | 2006-01-12 | 2007-01-12 | TiO2 nanostructures, membranes and films, and methods of making same |
| US11/653,192 | 2007-01-12 | ||
| PCT/US2007/078634 WO2008088588A2 (en) | 2007-01-12 | 2007-09-17 | Biocompatible scaffold for sensing proteins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101646401A CN101646401A (en) | 2010-02-10 |
| CN101646401B true CN101646401B (en) | 2012-05-23 |
Family
ID=41657978
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007800497563A Expired - Fee Related CN101646401B (en) | 2007-01-12 | 2007-09-17 | Biocompatible scaffold for sensing proteins |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101646401B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6503831B2 (en) * | 1997-10-14 | 2003-01-07 | Patterning Technologies Limited | Method of forming an electronic device |
| US6914279B2 (en) * | 2002-06-06 | 2005-07-05 | Rutgers, The State University Of New Jersey | Multifunctional biosensor based on ZnO nanostructures |
-
2007
- 2007-09-17 CN CN2007800497563A patent/CN101646401B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6503831B2 (en) * | 1997-10-14 | 2003-01-07 | Patterning Technologies Limited | Method of forming an electronic device |
| US6914279B2 (en) * | 2002-06-06 | 2005-07-05 | Rutgers, The State University Of New Jersey | Multifunctional biosensor based on ZnO nanostructures |
Non-Patent Citations (4)
| Title |
|---|
| D.V. Bavykin et al..A novel cation-binding TiO2 nanotube substrate for electro- and bioelectro-catalysis.《Electrochemistry Communications》.2005,第7卷1050-1058. * |
| Emmanuel Topoglidis et al.Direct electrochemistry and nitric oxide interaction of heme proteins adsorbed on nanocrystalline tin oxide electrodes.《Langmuir》.2003,第19卷(第17期),6894-6900. * |
| Thomas J.Webster et al..Increased osteoblast function on PLGA composites containing nanophase titania.《J Biomed Mater Res》.2005,第74A卷677-686. * |
| Wenjun Dong et al..Biocompatible nanofiber scaffolds on metal for controlled release and cell colonization.《Nanomedicine:Nanotechnology,Biology,and Medicine》.2006,第2卷248-252. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101646401A (en) | 2010-02-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jain et al. | Glycated hemoglobin detection with electrochemical sensing amplified by gold nanoparticles embedded N-doped graphene nanosheet | |
| Bartlett et al. | An enzyme switch employing direct electrochemical communication between horseradish peroxidase and a poly (aniline) film | |
| Bezbaruah et al. | Fabrication of anodically electrodeposited iridium oxide film pH microelectrodes for microenvironmental studies | |
| Babu et al. | Freestanding and binder free PVdF-HFP/Ni-Co nanofiber membrane as a versatile platform for the electrocatalytic oxidation and non-enzymatic detection of urea | |
| Liu et al. | Application of colloidal gold in protein immobilization, electron transfer, and biosensing | |
| Pejcic et al. | Impedance spectroscopy: Over 35 years of electrochemical sensor optimization | |
| Zuaznabar-Gardona et al. | A wide-range solid state potentiometric pH sensor based on poly-dopamine coated carbon nano-onion electrodes | |
| Yadav et al. | Tri-enzyme functionalized ZnO-NPs/CHIT/c-MWCNT/PANI composite film for amperometric determination of creatinine | |
| O’Hare et al. | Metal–metal oxide pH sensors for physiological application | |
| CN103336043B (en) | Preparation method of hydrogen peroxide biosensor | |
| Zhang et al. | An enzymatic glucose biosensor based on a glassy carbon electrode modified with manganese dioxide nanowires | |
| CN106525943B (en) | A kind of surface protein imprints construction method and its application of self energizing biological fuel cell sensor | |
| CN102735734A (en) | Non-invasive glucose sensor | |
| Mai et al. | Electrochemical determination of paraquat using a DNA-modified carbon ionic liquid electrode | |
| Thakur et al. | Highly sensitive enzymatic biosensor based on polyaniline-wrapped titanium dioxide nanohybrid for fish freshness detection | |
| Hathoot et al. | Simultaneous determination of ascorbic acid, uric acid and dopamine at modified electrode based on hybrid nickel hexacyanoferrate/poly (1, 5-diaminonaphthalene) | |
| Lou et al. | A 3D bio-platform constructed by glucose oxidase adsorbed on Au nanoparticles assembled polyaniline nanowires to sensitively detect glucose by electrochemiluminescence | |
| US10006880B2 (en) | Test strips having ceria nanoparticle electrodes | |
| Mazurenko et al. | Controlled electrochemically-assisted deposition of sol− gel biocomposite on electrospun platinum nanofibers | |
| Devnani et al. | Nanocomposite modified electrochemical sensor for sensitive and selective determination of noradrenaline | |
| Bracamonte et al. | Quaternized chitosan as support for the assembly of gold nanoparticles and glucose oxidase: Physicochemical characterization of the platform and evaluation of its biocatalytic activity | |
| Wu et al. | Sodium dodecyl sulfate doping PEDOT to enhance the performance of neural microelectrode | |
| Afkhami et al. | Electrochemical sensor for Dapsone using molecularly imprinted polypyrrole membrane as a recognition element | |
| Hu et al. | Fabrication and characterization of submicrometer-and nanometer-sized double-barrel pipets | |
| CN101646401B (en) | Biocompatible scaffold for sensing proteins |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: COUNCIL OF ARKANSAS STATE UNIVERSITY Free format text: FORMER OWNER: TECHNOLOGY DEVELOPMENT FOUNDATION OF ARKANSAS STATE UNIVERSITY Effective date: 20110920 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20110920 Address after: American Arkansas Applicant after: Univ Arkansas Technology Dev F. Address before: American Arkansas Applicant before: Univ Arkansas Technology Dev F. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120523 Termination date: 20130917 |