CN101424655B - Electrochemical tongs for developing protein molecule electronic device and method for producing the same - Google Patents
Electrochemical tongs for developing protein molecule electronic device and method for producing the same Download PDFInfo
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- CN101424655B CN101424655B CN2007101564938A CN200710156493A CN101424655B CN 101424655 B CN101424655 B CN 101424655B CN 2007101564938 A CN2007101564938 A CN 2007101564938A CN 200710156493 A CN200710156493 A CN 200710156493A CN 101424655 B CN101424655 B CN 101424655B
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Abstract
The invention relates to a pair of electrochemical pliers used for developing a protein molecule electronic device and a fabricating method thereof. The fabricating method is characterized in that the surface of an insulating basal material is uniformly coated with an inner layer and an outer layer of annular conductors of nanometer-level thickness, and a nanometer isolating layer of adjustable thickness is used for isolation. The annular conductors of the inner layer and the outer layer are respectively supplied with proper voltage, and electrostatic attracting force is used for clamping protein molecules causing the protein molecules to be arranged in parallel in a fixed direction; an outer circuit is used for supplying an activating signal to the protein molecules clamped by the electrochemical pliers; the relationship between an input signal and an output signal is analyzed to obtain the inner electronic transferring system of the protein molecules and develop the electronic device of the protein molecules. The electrochemical pliers provided by the invention has light action force, the shape and the quality of the protein can not easily change, touched molecule number is high, the signal is strong, the surface regeneration is easy, the used instrument is simple, the information is on-site, and the scanning speed is high. The fabricating method can be used for developing the electronic device of the protein molecules, particularly the electronic device of the protein molecules with high-speed response ability.
Description
Technical field
The present invention relates to Electroanalytical Chemistry, molecular electronic device, biosensor technology field, be specifically related to structure of a kind of electrochemical tongs that can be used for developing protein molecule electronic device and preparation method thereof.
Background technology
On a large scale, the development trend of VLSI (very large scale integrated circuits) is the size of constantly dwindling electron device, got into the nano electron device epoch at present.The development nano electron device mainly contains two approach at present: the one, and adopt microelectronic processing technique further to dwindle the conditional electronic size of devices; But near or when reaching nanoscale, quantum mechanical effects and thermodynamics statistical fluctuation effect make contemporary microelectronic development receive serious restriction gradually; The 2nd, the exploitation molecular electronic device.The latter is a frontier; But caused great attention both domestic and external; Numerous and confused drop into great amount of manpower and material resources, financial resources are researched and developed, this be because: compare with the conditional electronic device, the superiority of molecular electronic device is significant; Switch and logical operation are carried out on molecular level, and its volume can dwindle hundreds of and even thousands of times; Molecule can act on all directions in space, only is formed on the two-dimensional surface unlike traditional silicon integrated circuit, and molecule logical circuit closeness will improve about 100,000 times than conditional electronic device; Utilize chemical reaction to carry out work, only need little energy just can carry out; Possibility of collision is minimum between mobile electron, can not produce heating problem.Utilize these molecular electronic devices, can design and manufacture " biocomputer " of " molecular motor " and " biological missile " that the nanometer space is operated, 21st century novel concept.Molecular electronic device has two sources: the one, and synthetic macromolecule; The 2nd, the big molecule of natural biological, from the popularity in source, the superiority of performance, the latter has more advantage, and it is important that protein molecule wherein shows especially.There is a plurality of redox center protein molecule inside; Between the redox center with and have electron transport behavior widely with the outside; Mutual relationship and the electronics of exploring electron transfer center in this molecule shift and resident rule betwixt; Can find the characteristic of " molecular electronic device " that it has; Be that protein molecule excites down in appropriate chemi-excitation, photochemical excitation, galvanochemistry, have the character of conditional electronic devices such as being similar to resistance, electric capacity, diode, switch, lead, logic gate, storer, feedback circuit, realize the protein molecule electronic device function.
The exploitation molecular electronic device adopts method (Appl.Phys.Lett., 2002, the 81:3043 of PSTM (STM) or conductive pinpoint atomic force microscope (CP-AFM) more at present; Science, 2001,294:571; Science, 2001,292:2303; Science, 2003,301:1221; J.Am.Chem Soc., 2003,125:16164), few molecules is embedded between two electrodes, (metal-molecules-metal, molecule knot MMM) applies excitation signal, analyzes its response to form metal-molecule-metal.But come with some shortcomings with this method exploitation protein molecule electronic device: 1. desire realizes the connection of MMM, must use contact mode, and electrode or conductive pinpoint directly contact with protein molecule, is prone to cause protein distortion sex change, and then influences its electrochemical properties; 2. it is little to contact the molecule number, and weak output signal is subject to noise; 3. the Surface Renewal of electrode or conductive pinpoint is relatively more difficult; 4. employed instrument costs an arm and a leg, and research cost is high.For remedying above-mentioned deficiency, (Nature, 1992,356:361 such as Armstrong; J.Am.Chem.Soc., 2005,127:14964; Biochemistry, 2003,42:8653; Curr.Opin.Chem.Biol.; 2005; 9:110) propose to adopt the protein film voltammetry that redox protein matter is immobilized in the electrode surface of friendly interface with individual layer or inferior form of single sheet; Apply the current potential disturbance and cause electronics to import, spread out of protein or portion migration within it into, with these electron transfer behaviors of cyclic voltammetric technical research and current potential, isoparametric relation of time, the molecular electronic device behavior of investigation protein; This method adopts the immobilized protein of intermolecular force, and acting force is little and protein is not yielding; Contact molecule number is more, and signal is stronger; Electrode surface upgrades relatively easy, and polishing gets final product; Employed instrument is simple.But this method also comes with some shortcomings simultaneously: conclusion is come out through the galvanochemistry information inference, is indirect, non-at-scene; The electrochemical techniques that adopted are mainly the cyclic voltammetric technology, and the information that can provide is limited, still can not satisfy the demand of the various molecular electronic devices of exploitation; The operable sweep velocity of cyclic voltammetric technology is less, even if adopt the fastest known ultrafast voltammetry at present, also only has about 1MHz, can't develop the protein molecule electronic device with high-speed response ability.
To above-mentioned two kinds of methods deficiency separately, the present invention has made a kind of electrochemical tongs of developing protein molecule electronic device, the slight and not yielding sex change of protein of acting force; Contact molecule number is many, and signal is stronger; Surface Renewal is easy, and the cut-out front end forms new cross section and gets final product; Employed instrument is simple; Information is on-the-spot; Electrochemical techniques are various; Sweep velocity is fast, can develop the protein molecule electronic device with high-speed response ability.Electrochemical tongs of above-mentioned exploitation protein molecule electronic device and preparation method thereof is not all seen in other documents or patent.
Summary of the invention
Primary technical matters to be solved by this invention be overcome that the acting force that exists in two kinds of methods of above-mentioned existing exploitation protein molecule electronic device causes greatly that protein distortion sex change, signal are weak, Surface Renewal is difficult, instrument and equipment is expensive, information be non-at-scene, sweep velocity slow and can't develop the deficiency of the protein molecule electronic device of high-speed response ability; A kind of electrochemical tongs of developing protein molecule electronic device is provided, the slight and not yielding sex change of protein of its acting force; Contact molecule number is many, and signal is stronger; Surface Renewal is easy, and the cut-out front end forms new cross section and gets final product; Employed instrument is simple; Information is on-the-spot; Electrochemical techniques are various; Sweep velocity is fast, can develop the protein molecule electronic device with high-speed response ability.
Another technical matters to be solved by this invention is to overcome the acting force that exists in two kinds of methods of above-mentioned existing exploitation protein molecule electronic device to cause protein distortion sex change greatly; Signal a little less than; The Surface Renewal difficulty; Instrument and equipment is expensive; Information is non-at-scene; Sweep velocity can't be developed the deficiency of the protein molecule electronic device of high-speed response ability slowly; Provide a kind of simple and practical; Implement easily; Reuse; The method for making of the electrochemical tongs of exploitation protein molecule electronic device with low cost
The technical scheme that solves the primary technical matters of the present invention is such: a kind of electrochemical tongs of developing protein molecule electronic device; It is characterized in that: the inner conductor layer, the outer conductor layer that evenly are coated with nanometer grade thickness on the insulating substrate surface; They are isolated through the inner insulating layer of nanometer grade thickness; Inner conductor layer is connected lead separately with outer conductor layer with conducting resinl; The electric charge that utilizes the surperficial amino acid residue of protein molecule to have applies appropriate voltage respectively to vise protein molecule on inner conductor layer, outer conductor layer, it is aligned between inner conductor layer and outer conductor layer.
As improvement, described inner conductor layer, outer conductor layer are done circlewise, ring conductor, outer shroud conductor in becoming, and inner insulating layer is also done circlewise, and thickness is adjustable, is convenient to actual manufacturing like this.
As improvement; Outside inner insulating layer, set up glass capillary in addition between ring conductor, the outer shroud conductor epimere in described; The lead of ring conductor is fixed on the insulating substrate through epoxy resin in making; And the lead of outer shroud conductor is fixed on the glass capillary through epoxy resin, makes lead be more prone to be arranged on the electrochemical tongs.
As improvement, the outer shroud conductive surface is external insulation layer in even coating in glass capillary bottom or the polymerization at least, plays necessary protective effect.
As preferably, insulating substrate is spun glass, quartz glass fibre, ceramic fiber or fibrous asbestos, is convenient to actual use.
As preferably, interior ring conductor, outer shroud conductor by carbon, comprise that the precious metal material or the Rare Metals Materials of gold, platinum, silver constitute, make electrochemical tongs keep excellent conducting performance, improve the work sensitivity;
Further improve, the method for evaporation or sputter is adopted in the coating of interior ring conductor, outer shroud conductor, and its thickness is regulated through the time and the intensity that change evaporation or sputter, be controlled between the 1-50nm, and so easy realization, method is practical.
Further improve, inner insulating layer, external insulation layer are made up of the high molecular polymer that electropolymerization forms, and the thickness of said inner insulating layer, external insulation layer was regulated through the current potential and the time that change electropolymerization, was controlled between the 1-500nm.So easy realization, method is simple and practical.
The technical scheme that solves another technical matters of the present invention is such: a kind of electrochemical tongs method for making of developing protein molecule electronic device is characterized in that comprising the steps:
A) evenly plate one deck conductor on the insulating substrate surface, ring conductor in constituting encircles conductor layer and is drawn by lead through conducting resinl in this;
B) in this, behind even coating of ring conductor one end surfaces or the polymerization one nano level inner insulating layer, again the other end is sealed in the glass capillary, fixes with epoxy resin;
C) evenly plate one deck conductor at this inner insulating layer and glass capillary outside surface, constitute the outer shroud conductor, this outer shroud conductor layer is drawn by lead through conducting resinl, fixes with epoxy resin;
D) this outer shroud conductive surface even coating or polymerization one external insulation layer from the glass capillary bottom at least;
E) will be coated with vertical cut-out of insulating substrate front end of conductor layer and insulation course, two-layer nanometer grade thickness ring-shaped conductor cross section is as electrochemical tongs inside and outside exposing.
Said insulating substrate is spun glass, quartz glass fibre, ceramic fiber, fibrous asbestos; Ring conductor, outer shroud conductor are made up of carbon, precious metal material gold, platinum, silver or Rare Metals Materials in said; The method of evaporation or sputter is adopted in the coating of ring conductor, outer shroud conductor in said, and its thickness is regulated through the time and the intensity that change evaporation or sputter, is controlled between the 1-50nm; Said inner insulating layer, external insulation layer are made up of the high molecular polymer that electropolymerization forms, and the thickness of inner insulating layer, external insulation layer was regulated through the current potential and the time that change electropolymerization, was controlled between the 1-500nm.
Compared with prior art, advantage of the present invention has following technique effect:
(1) the present invention utilizes the electric charge that the amino acid residue on protein molecule surface has; On inside and outside two-layer ring-shaped conductor, apply appropriate voltage respectively to vise protein molecule; It is aligned between inside and outside two-layer ring-shaped conductor with the exploitation protein molecule electronic device; Acting force is mainly electrostatic attraction, slightly and not can cause protein distortion sex change;
(2) the present invention is owing to realize more protein molecule and between inside and outside two-layer ring-shaped conductor, align with parallel form, thereby signal is stronger;
(3) electrochemical tongs Surface Renewal of the present invention is very easy to, and unsalted surface is exposed in the vertical cut-out of its front end get final product, and is simple to operate, good stability;
(4) the present invention is used to develop protein molecule electronic device; The instrument simple, inexpensive of required use; Only need AWG, oscillograph, amplifier to build a system, about 1/50 of only about STM of total price or CF-AFM price, and should the research system need not operating cost;
(5) information of the resulting protein molecule electronic device of the present invention is direct, on-the-spot;
(6) the present invention can use various waveforms to study as excitation signal, and abundant information is various;
(7) the present invention can use high-frequency input signal, develops the protein molecule electronic device with high-speed response ability;
(8) electrochemical tongs manufacture craft of the present invention is simple, practical, processing ease, and manufacturing conditions is controlled easily, and is with low cost, all can make at general chemical laboratory, has application value preferably.
Electrochemical tongs provided by the invention can vise protein molecule through electrostatic attraction, and its orientation is arranged in parallel between inside and outside two-layer ring-shaped conductor with the exploitation protein molecule electronic device; Slight and the not yielding sex change of protein of acting force; Contact molecule number is many and signal is stronger, and Surface Renewal is easy, and employed instrument is simple; Information is on-the-spot; Sweep velocity is fast, can be used for developing protein molecule electronic device, particularly has the protein molecule electronic device of high-speed response ability.
Description of drawings
Fig. 1 a is the structure cut-open view (M) of electrochemical tongs of the present invention;
Fig. 1 b is the structure cross-sectional view (N) of electrochemical tongs of the present invention;
Fig. 2 aligns synoptic diagram for protein molecule on the electrochemical tongs xsect;
Fig. 3 is for adopting the Circuits System synoptic diagram of electrochemical tongs exploitation protein molecule electronic device;
Fig. 4 is the I～V curve of CER on electrochemical tongs.
Among its Fig. 1:
1, insulating substrate, 2, interior ring conductor, 3, the outer shroud conductor, 4, inner insulating layer, 5, conducting resinl, 6, lead, 7, glass capillary, 8, epoxy resin, 9, conducting resinl, 10, lead, 11, epoxy resin, 12, external insulation layer; M, longitudinal profile, N, xsect.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment 1
Select for use spun glass as insulating substrate in the present embodiment; Gold is as interior ring conductor 2, outer shroud conductor 3; Clean with ethanol, distilled water ultrasonic cleaning, conducting resinl 5,9 adopts silver conductive adhesive, the high molecular polymer that inside and outside insulation course 4,12 adopts polymerization methods to form.Its concrete method for making is: place acetone to reflux 10 hours spun glass earlier; Use ethanol, distilled water ultrasonic cleaning again, behind the bone dry intercepting 3-4cm length as insulating substrate 1, its surface evaporation plate the thick gold of the about 20nm of one deck serve as in the ring conductor 2; The copper wire that is adhered to Φ 0.1mm with silver conductive adhesive 5 is terminal; Ring conductor 6 in constituting is immersed in the spun glass of this surface gold-plating in phenol and the 2-allyl benzene phenol solution, and gold layer is kept the about 3-6min of constant potential 5.2V; Make phenol and 2-chavicol in golden laminar surface generation polymerization, form the inner insulating layer 4 that thickness is about 5-10nm through 150 ℃ of high-temperature bakings; Said structure is carefully inserted glass capillary 7; With the glass capillary drawing device it is sealed; An end of ring conductor 2 and inner insulating layer 4 is exposed to outside the glass capillary 7 in being coated with, and the copper conductor that the other end exposes is fixing with epoxy resin 8, has constituted interior ring conductor 6; Adopt disposal route same as described above, evaporation plates the thick gold of the about 20nm of one deck and serves as outer shroud conductor 3 with glass capillary 7 outer walls at inner insulating layer 4, draws through the copper wire of silver conductive adhesive 9 usefulness Φ 0.1mm, as outer shroud conductor 10, fixes with epoxy resin 11; Adopt disposal route same as described above, form the external insulation layer 12 that thickness is about 10-20nm in outer shroud conductor 3 surfaces again; At last, cut off, expose the cross section of inside and outside ring-shaped conductor 2,3, promptly obtain electrochemical tongs with insulating substrate 1 front end of scalpel with spun glass.
The above-mentioned electrochemical tongs that makes is immersed in the CER hac buffer (pH=4.64) of 5mmol/L, ring conductor constant potential is-0.1V for+0.1V, outer shroud conductor constant potential in keeping, after 2 hours; Taking-up is washed with ultrapure water, and drying is pressed accompanying drawing 3 and installed; AWG send linear ramp-0.1V～+ 0.1V; Sweep speed for 1V/s, input signal, output signal input oscillograph are respectively X axle, Y axle, record current-voltage (curve of I～V); Shown in accompanying drawing 4, be illustrated in the molecular electronic device character that CER under this experiment condition shows as resistance.
Explained on interior ring conductor 2, outer shroud conductor 3 to apply appropriate voltage respectively, its orientation is arranged in parallel between interior ring conductor 2 and outer shroud conductor 3 to vise protein molecule.Apply excitation signal in the protein molecule that is vised by electrochemical tongs through external circuit, analyze the relation between input signal and the output signal, know the electron transport mechanism that protein molecule is inner, the exploitation protein molecule electronic device.
Claims (10)
1. electrochemical tongs of developing protein molecule electronic device; It is characterized in that: the inner conductor layer, the outer conductor layer that evenly are coated with nanometer grade thickness on the insulating substrate surface; They are isolated through the inner insulating layer of nanometer grade thickness; Inner conductor layer is connected lead separately with outer conductor layer with conducting resinl; The electric charge that utilizes the surperficial amino acid residue of protein molecule to have applies appropriate voltage respectively to vise protein molecule on inner conductor layer, outer conductor layer, it is aligned between inner conductor layer and outer conductor layer.
2. electrochemical tongs according to claim 1 is characterized in that: inner conductor layer, outer conductor layer are done circlewise, ring conductor, outer shroud conductor in becoming, and inner insulating layer is also done circlewise, and thickness is adjustable.
3. electrochemical tongs according to claim 2; It is characterized in that: outside inner insulating layer, set up glass capillary in addition between interior ring conductor, the outer shroud conductor epimere; The lead of ring conductor is fixed on the insulating substrate through epoxy resin in making, and the lead of outer shroud conductor is fixed on the glass capillary through epoxy resin.
4. electrochemical tongs according to claim 3 is characterized in that: the outer shroud conductive surface is external insulation layer in even coating in glass capillary bottom or the polymerization at least.
5. electrochemical tongs according to claim 4 is characterized in that: insulating substrate is spun glass, ceramic fiber or fibrous asbestos.
6. be pursuant to the described electrochemical tongs of claim 4, it is characterized in that: interior ring conductor, outer shroud conductor by carbon, comprise that the precious metal material or the Rare Metals Materials of gold, platinum, silver constitute.
7. electrochemical tongs according to claim 4 is characterized in that: the method for evaporation or sputter is adopted in the coating of interior ring conductor, outer shroud conductor, and its thickness is regulated through the time and the intensity that change evaporation or sputter, is controlled between the 1-50nm.
8. electrochemical tongs according to claim 4; It is characterized in that: inner insulating layer, external insulation layer are made up of the high molecular polymer that electropolymerization forms; The thickness of said inner insulating layer, external insulation layer was regulated through the current potential and the time that change electropolymerization, was controlled between the 1-500nm.
9. an electrochemical tongs method for making of developing protein molecule electronic device is characterized in that comprising the steps:
A) evenly plate one deck conductor on the insulating substrate surface, ring conductor in constituting encircles conductor layer and is drawn by lead through conducting resinl in this;
B) in this, behind even coating of ring conductor one end surfaces or the polymerization one nano level inner insulating layer, again the other end is sealed in the glass capillary, fixes with epoxy resin;
C) evenly plate one deck conductor at this inner insulating layer and glass capillary outside surface, constitute the outer shroud conductor, this outer shroud conductor layer is drawn by lead through conducting resinl, fixes with epoxy resin;
D) this outer shroud conductive surface even coating or polymerization one external insulation layer from the glass capillary bottom at least;
E) will be coated with vertical cut-out of insulating substrate front end of conductor layer and insulation course, two-layer nanometer grade thickness ring-shaped conductor cross section is as electrochemical tongs inside and outside exposing.
10. electrochemical tongs method for making according to claim 9 is characterized in that: said insulating substrate is spun glass, ceramic fiber, fibrous asbestos; Ring conductor, outer shroud conductor are made up of carbon, precious metal material gold, platinum, silver or Rare Metals Materials in said; The method of evaporation or sputter is adopted in the coating of ring conductor, outer shroud conductor in said, and its thickness is regulated through the time and the intensity that change evaporation or sputter, is controlled between the 1-50nm; Said inner insulating layer, external insulation layer are made up of the high molecular polymer that electropolymerization forms, and the thickness of inner insulating layer, external insulation layer was regulated through the current potential and the time that change electropolymerization, was controlled between the 1-500nm.
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