CN105929009A - Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal - Google Patents
Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal Download PDFInfo
- Publication number
- CN105929009A CN105929009A CN201610223437.0A CN201610223437A CN105929009A CN 105929009 A CN105929009 A CN 105929009A CN 201610223437 A CN201610223437 A CN 201610223437A CN 105929009 A CN105929009 A CN 105929009A
- Authority
- CN
- China
- Prior art keywords
- graphene
- biofilm
- silk polypeptide
- solution
- preparation
- 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.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Abstract
The invention relates to the field of graphene biotechnological application, and concretely relates to a preparation method of a silk polypeptide/graphene biofilm used for detecting biological signal. The method includes the following steps: 1) providing aqueous solution of graphene oxide and silk polypeptide solution; 2) adding sodium hydroxide and pyrenebutyric acid into the aqueous solution of graphene oxide; 3) adding hydrazine hydrate solution, heating the mixture to perform reduction; 4) cooling the mixture to room temperature, performing centrifugation, pouring the supernatant on a filter paper, performing pumping filtration to obtain a film; 5) repeating the steps 2), 3), and 4) to obtain a multilayer film; 6) washing the film with water and drying the film; 7) putting the film in a mixed solution containing EDC and NHS to soak the film, and washing the film with water; and 8) dropping the silk polypeptide solution on the film to perform complete reaction to obtain the silk polypeptide/graphene biofilm, washing the biofilm with PBS buffer several times, adjusting the pH value, and air drying the biofilm.
Description
Technical field
The present invention relates to the application of graphene film, the preparation of a kind of Graphene biofilm for detecting bio signal.
Background technology
Graphene is a kind of to be formed the flat film that hexangle type is honeycomb lattice, the two-dimensional material of only one of which carbon atom thickness by carbon atom with sp2 hybrid orbital.Graphene be the thinnest in the world be the hardest nano material also, it is almost fully transparent, only absorb the light of 2.3%, heat conductivity is up to 5300 W/m K, higher than CNT and diamond, under room temperature, its electron mobility is more than 15000 cm2/V s, again ratio CNT or silicon wafer heights, and resistivity only about 10-6 Ω cm, lower than copper or silver.The appearance of Graphene breaches the conventional understanding to two-dimensional material, and it has now been discovered that Graphene has unusual electric conductivity, beyond the intensity of iron and steel decades of times and fabulous light transmission, its appearance is more expected to cause one to take turns revolution at hyundai electronics sciemtifec and technical sphere.In Graphene, electronics can extremely efficiently migrate, and traditional quasiconductor and conductor, such as silicon and copper are far from, Graphene shows well.Due to electronics and the collision of atom, the form of traditional quasiconductor and conductor heat releases some energy, and general computer chip wastes the electric energy of 72%-81% by this way, and Graphene is the most different, its electron energy will not be depleted, and this makes it be provided with the good characteristic being not of the common run.Construction features additionally, due to Graphene, it is easy to form the interaction between π-π, can easily carry out functional modification and be widely used in the field such as living cells detection and medicine transmission of biological field.
Summary of the invention
It is an object of the invention to provide the preparation method of a kind of Graphene biofilm for detecting bio signal, this preparation method thin film that is easy and that prepare is soft, has higher using value.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal of the present invention, comprises the steps of
1) aqueous solution and the silkworm silk polypeptide solution of graphene oxide are provided；
2) to step 1) described in graphene oxide water solution in add the sodium hydroxide of certain mass and pyrene butanoic acid obtains mixed solution；
3) to step 2) described in mixed solution in add hydrazine hydrate solution, add thermal reduction at a certain temperature；
4) solution after reduction is cooled to room temperature, is then centrifuged for obtaining supernatant, supernatant is poured on filter paper and carries out sucking filtration, thin film can be obtained；
5) repeat the above steps 2), 3) and 4), plural layers can be obtained；
6) it is dried 4 ~ 8 hours by thin film deionized water rinsing and under 40 ~ 60 degrees Celsius, pyrene butanoic acid/graphene film can be obtained；
7) by step 6) pyrene butanoic acid/graphene film of obtaining is placed with in the mixed solution of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) and N-hydroxy-succinamide sulfonate sodium (NHS) immersion, obtains, with deionized water rinsing, pyrene butanoic acid/graphene film that EDC/NHS connects after taking-up；
7) drip to silkworm silk polypeptide solution, on pyrene butanoic acid/graphene film that EDC/NHS connects, fully react 1 ~ 2 hour, obtain silkworm silk polypeptide/Graphene biofilm, then dry after biofilm PBS is repeatedly rinsed regulation pH value.
Preferential, described step 1) the concentration of graphene oxide water solution be 0.1 ~ 10mg/mL；The concentration of silkworm silk polypeptide is 0.1 ~ 10mg/mL.
Preferential, described step 1) the preparation method of graphene oxide include:
By graphite powder and 0.5 ~ 20g sodium nitrate mixing of 1 ~ 10g, under stirring, it is slowly added to the concentrated sulphuric acid of 20 ~ 50mL, ultrasonic 15 ~ 30 minutes, forms suspension；
Under condition of ice bath, suspension is placed on magnetic stirring apparatus continuation stirring 30 minutes, then 3 ~ 30g potassium permanganate is slowly added in suspension, continue to keep magnetic agitation 10 ~ 14 hours under condition of ice bath；
After withdrawing condition of ice bath, then instill the deionized water of 20 ~ 50mL, maintain the temperature at and under the water bath condition of 40 ~ 50 degrees Celsius, continue stirring 10 ~ 14 hours；
It is adjusted to temperature in the water-bath of 30 ~ 38 degrees Celsius continue stirring 10 ~ 14 hours；
Again toward the hydrogenperoxide steam generator of add 10 ~ 30mL in solution 30%, continue the stirred in water bath 2 ~ 4 hours at 30 ~ 38 degrees Celsius；
Carry out repeatedly eccentric cleaning after adding dilute hydrochloric acid, add water and be centrifuged cleaning, adjust pH value to neutrality, adjustment graphene oxide concentration.
Preferential, described step 1) the preparation method of silkworm silk polypeptide solution include:
Bombyx bombycis is boiled 20 ~ 40min, at the NaCO of 0.2 ~ 0.3M3Middle removal sericin, then cleans with secondary water, is dried more than 12 hours afterwards, is dissolved in the LiBr solution of 9 ~ 11M and is incubated 3 ~ 4h at a temperature of 55 ~ 65 DEG C.Loading in the bag filter of MWCO3500 and dialyse 2 ~ 3 days, period only stirs on magnetic stirring apparatus and does not heats.
Preferential, described step 2) mixed solution in the concentration of sodium hydroxide and pyrene butanoic acid be respectively 4 ~ 40mg/mL and 5 ~ 50mg/mL.
Preferential, described step 3) in the mass concentration of hydrazine hydrate solution be 60 ~ 90%, the heated at constant temperature time is 1 ~ 3 hour, and heating-up temperature is 60 ~ 80 degrees Celsius.
Preferential, described step 4) centrifugal rotational speed be 6000 ~ 12000rpm, the centrifugal time is 10 ~ 20 minutes, and filter paper aperture is 30 ~ 50 microns.
Preferential, described step 7) in EDC and NHS mixed solution in the molar concentration of EDC be 3 ~ 6 mol/mL, the molar concentration of NHS is 6 ~ 18 mol/mL, and soak time is 0.5 ~ 2 hour.
Accompanying drawing explanation
In order to make the purpose of the present invention, technical scheme and beneficial effect become apparent from, the present invention provides drawings described below to illustrate:
Fig. 1 is the one layer of silkworm silk polypeptide/Graphene biofilm of embodiment 1 preparation.
Fig. 2 is the five layers of silkworm silk polypeptide/Graphene biofilm of embodiment 2 preparation.
Fig. 3 is that the graphene/silk biomembrane of embodiment 2 utilizes the bio signal of NO in the detection cell of cyclic voltammetry.
Detailed description of the invention
The preferred embodiments of the present invention will be described in detail below.
Embodiment
1
1) sodium hydroxide and the 120mg pyrene butanoic acid that add 80mg in the graphene oxide water solution of the 0.4mg/ml of 20ml obtain mixed solution；
2) to step 1) described in mixed solution in add 200 microlitre hydrazine hydrate solutions, be maintained under 80 degree celsius temperature and add thermal reduction 1.5 hours；
3) solution after reduction is cooled to room temperature, is then centrifuged for obtaining supernatant；
4) take supernatant to be poured on filter paper and carry out sucking filtration, obtain thin film；
5) it is dried 4 ~ 8 hours by thin film deionized water rinsing and under 40 ~ 60 degrees Celsius, pyrene butanoic acid/graphene film can be obtained；
6) by step 5) pyrene butanoic acid/graphene film of obtaining is placed with in the mixed solution of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) and N-hydroxy-succinamide sulfonate sodium (NHS) immersion 1h, wherein the molar concentration of EDC is 5 mol/mL, the molar concentration of NHS is 10 mol/mL, obtains, with deionized water rinsing, pyrene butanoic acid/graphene film that EDC/NHS connects after taking-up；
7) silkworm silk polypeptide solution is dripped on pyrene butanoic acid/graphene film that EDC/NHS connects, fully reaction 1.5 hours, obtain silkworm silk polypeptide/Graphene biofilm, then dry after biofilm PBS is repeatedly rinsed regulation pH value.
Fig. 1 is the one layer of silkworm silk polypeptide/Graphene biofilm of embodiment 1 preparation.
Embodiment
2
1) sodium hydroxide and the 600mg pyrene butanoic acid that add 400mg in the graphene oxide water solution of the 0.4mg/ml of 100ml obtain mixed solution；
2) to step 1) described in mixed solution in add 1000 microlitre hydrazine hydrate solutions, be maintained under 80 degree celsius temperature and add thermal reduction 1.5 hours；
3) solution after reduction is cooled to room temperature, is then centrifuged for obtaining supernatant；
4) take 20ml supernatant to be poured on filter paper and carry out sucking filtration, obtain thin film, by thin film deionized water rinsing；
5) repeat the above steps 4) four times, obtain 5 layer films；
6) thin film is dried 4 ~ 8 hours under 40 ~ 60 degrees Celsius, 5 layers of pyrene butanoic acid/graphene film can be obtained；
7) by step 6) pyrene butanoic acid/graphene film of obtaining is placed with in the mixed solution of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) and N-hydroxy-succinamide sulfonate sodium (NHS) immersion 1h, wherein the molar concentration of EDC is 5 mol/mL, the molar concentration of NHS is 10 mol/mL, obtains, with deionized water rinsing, pyrene butanoic acid/graphene film that EDC/NHS connects after taking-up；
8) silkworm silk polypeptide solution is dripped on pyrene butanoic acid/graphene film that EDC/NHS connects, fully reaction 1.5 hours, obtain silkworm silk polypeptide/Graphene biofilm, then dry after biofilm PBS is repeatedly rinsed regulation pH value.
Fig. 2 is the five layers of silkworm silk polypeptide/Graphene biofilm of embodiment 2 preparation.
Fig. 3 is that the graphene/silk biomembrane of embodiment 2 utilizes the bio signal of NO in the detection cell of cyclic voltammetry.Curve 1 and curve 2 respectively do not have NO and have the curve under NO RST.It can be seen that biomembrane can be utilized to detect the NO signal of cell release.
Finally illustrate is, preferred embodiment above is only in order to illustrate technical scheme and unrestricted, although the present invention being described in detail by above-mentioned preferred embodiments, but skilled artisan would appreciate that, in the form and details it can be made various change, without departing from claims of the present invention limited range.
Claims (8)
1. for detecting the preparation method of the silkworm silk polypeptide/Graphene biofilm of bio signal, it is characterised in that comprise the steps of
1) aqueous solution and the silkworm silk polypeptide solution of graphene oxide are provided；
2) to step 1) described in graphene oxide water solution in add the sodium hydroxide of certain mass and pyrene butanoic acid obtains mixed solution；
3) to step 2) described in mixed solution in add hydrazine hydrate solution, add thermal reduction at a certain temperature；
4) solution after reduction is cooled to room temperature, is then centrifuged for obtaining supernatant, supernatant is poured on filter paper and carries out sucking filtration, thin film can be obtained；
5) repeat the above steps 2), 3) and 4), plural layers can be obtained；
6) it is dried 4 ~ 8 hours by thin film deionized water rinsing and under 40 ~ 60 degrees Celsius, pyrene butanoic acid/graphene film can be obtained；
7) by step 6) pyrene butanoic acid/graphene film of obtaining is placed with in the mixed solution of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) and N-hydroxy-succinamide sulfonate sodium (NHS) immersion, obtains, with deionized water rinsing, pyrene butanoic acid/graphene film that EDC/NHS connects after taking-up；
7) drip to silkworm silk polypeptide solution, on pyrene butanoic acid/graphene film that EDC/NHS connects, fully react 1 ~ 2 hour, obtain silkworm silk polypeptide/Graphene biofilm, then dry after biofilm PBS is repeatedly rinsed regulation pH value.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterised in that described step 1) the concentration of graphene oxide water solution be 0.1 ~ 10mg/mL；The concentration of silkworm silk polypeptide is 0.1 ~ 10mg/mL.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterised in that described step 1) the preparation method of graphene oxide include:
By graphite powder and 0.5 ~ 20g sodium nitrate mixing of 1 ~ 10g, under stirring, it is slowly added to the concentrated sulphuric acid of 20 ~ 50mL, ultrasonic 15 ~ 30 minutes, forms suspension；
Under condition of ice bath, suspension is placed on magnetic stirring apparatus continuation stirring 30 minutes, then 3 ~ 30g potassium permanganate is slowly added in suspension, continue to keep magnetic agitation 10 ~ 14 hours under condition of ice bath；
After withdrawing condition of ice bath, then instill the deionized water of 20 ~ 50mL, maintain the temperature at and under the water bath condition of 40 ~ 50 degrees Celsius, continue stirring 10 ~ 14 hours；
It is adjusted to temperature in the water-bath of 30 ~ 38 degrees Celsius continue stirring 10 ~ 14 hours；
Again toward the hydrogenperoxide steam generator of add 10 ~ 30mL in solution 30%, continue the stirred in water bath 2 ~ 4 hours at 30 ~ 38 degrees Celsius；
Carrying out repeatedly eccentric cleaning after adding dilute hydrochloric acid, add water and be centrifuged cleaning, adjustment pH value is to neutral, then adjusts graphene oxide concentration.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterised in that described step 1) the preparation method of silkworm silk polypeptide solution include:
Bombyx bombycis is boiled 20 ~ 40min, at the NaCO of 0.2 ~ 0.3M3Middle removal sericin, then cleans with secondary water, is dried more than 12 hours afterwards, is dissolved in the LiBr solution of 9 ~ 11M and is incubated 3 ~ 4h at a temperature of 55 ~ 65 DEG C；
Loading in the bag filter of MWCO3500 and dialyse 2 ~ 3 days, period only stirs on magnetic stirring apparatus and does not heats.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterised in that described step 2) mixed solution in the concentration of sodium hydroxide and pyrene butanoic acid be respectively 4 ~ 40mg/mL and 5 ~ 50mg/mL.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterized in that, described step 3) in the mass concentration of hydrazine hydrate solution be 60 ~ 90%, the heated at constant temperature time is 1 ~ 3 hour, and heating-up temperature is 60 ~ 80 degrees Celsius.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterized in that, described step 4) centrifugal rotational speed be 6000 ~ 12000rpm, the centrifugal time is 10 ~ 20 minutes, and filter paper aperture is 30 ~ 50 microns.
The preparation method of the silkworm silk polypeptide/Graphene biofilm for detecting bio signal the most according to claim 1, it is characterized in that, described step 7) in EDC and NHS mixed solution in the molar concentration of EDC be 3 ~ 6 mol/mL, the molar concentration of NHS is 6 ~ 18 mol/mL, and soak time is 0.5 ~ 2 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610223437.0A CN105929009A (en) | 2016-04-12 | 2016-04-12 | Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610223437.0A CN105929009A (en) | 2016-04-12 | 2016-04-12 | Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105929009A true CN105929009A (en) | 2016-09-07 |
Family
ID=56837972
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610223437.0A Pending CN105929009A (en) | 2016-04-12 | 2016-04-12 | Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105929009A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103173882A (en) * | 2013-04-11 | 2013-06-26 | 大连海事大学 | Method for preparing graphene/ silk composite fiber |
| CN104489922A (en) * | 2014-12-11 | 2015-04-08 | 上海烟草集团有限责任公司 | Preparation and application of graphene oxide-bonded silica gel composite |
| CN104707180A (en) * | 2015-02-06 | 2015-06-17 | 福州大学 | BMP loaded silk fibroin/collagen scaffold material and preparation method thereof |
| CN104998302A (en) * | 2015-08-31 | 2015-10-28 | 中原工学院 | Nano cartilage repair material taking aligned nanofiber mat as skeleton and preparation method of nano cartilage repair material |
| CN105061563A (en) * | 2015-07-28 | 2015-11-18 | 苏州普罗达生物科技有限公司 | Fibroin polypeptide and preparation and application thereof |
| CN105085958A (en) * | 2015-09-17 | 2015-11-25 | 李爱冰 | Polyphenol oxidase nano high-efficiency absorption fixed film based on modified silk protein and preparation method of film |
-
2016
- 2016-04-12 CN CN201610223437.0A patent/CN105929009A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103173882A (en) * | 2013-04-11 | 2013-06-26 | 大连海事大学 | Method for preparing graphene/ silk composite fiber |
| CN104489922A (en) * | 2014-12-11 | 2015-04-08 | 上海烟草集团有限责任公司 | Preparation and application of graphene oxide-bonded silica gel composite |
| CN104707180A (en) * | 2015-02-06 | 2015-06-17 | 福州大学 | BMP loaded silk fibroin/collagen scaffold material and preparation method thereof |
| CN105061563A (en) * | 2015-07-28 | 2015-11-18 | 苏州普罗达生物科技有限公司 | Fibroin polypeptide and preparation and application thereof |
| CN104998302A (en) * | 2015-08-31 | 2015-10-28 | 中原工学院 | Nano cartilage repair material taking aligned nanofiber mat as skeleton and preparation method of nano cartilage repair material |
| CN105085958A (en) * | 2015-09-17 | 2015-11-25 | 李爱冰 | Polyphenol oxidase nano high-efficiency absorption fixed film based on modified silk protein and preparation method of film |
Non-Patent Citations (2)
| Title |
|---|
| CHUN XIAN GUO ET AL.: "RGD-Peptide Functionalized Graphene Biomimetic Live-Cell Sensor for Real-Time Detection of Nitric Oxide Molecules", 《 ACS NANO》 * |
| WENJING YUAN ET AL.: "Ultrasensitive and Selective Nitrogen Dioxide Sensor Based on SelfAssembled Graphene/Polymer Composite Nanofibers", 《 ACS APPLIED MATERIALS & INTERFACES》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103203468B (en) | A kind of preparation method of nano silver wire | |
| CN106629834B (en) | A kind of method that recrystallization prepares lead halogen perovskite nano wire | |
| Xiao et al. | Encapsulation and regeneration of perovskite film by in Situ forming cobalt porphyrin polymer for efficient photovoltaics | |
| CN107275492A (en) | Introduce the method that nonsolute bromide compound additive prepares mixed halogen perovskite | |
| CN105084858B (en) | A kind of preparation method of graphene film | |
| CN104787751B (en) | A kind of graphene powder and preparation method thereof | |
| CN107663648B (en) | A kind of intercalation molybdenum oxide single crystal film and its preparation method and application | |
| CN106025067A (en) | Method for generating perovskite film through solution method and device application thereof | |
| CN104876211A (en) | Method for preparing graphene by rotationally shearing in electric field environment | |
| CN106058060A (en) | Method for preparing high-quality perovskite crystal thin film | |
| CN106702732A (en) | Graphene-copper composite fiber and preparation method thereof | |
| CN105929009A (en) | Preparation method of silk polypeptide/graphene biofilm used for detecting biological signal | |
| CN102936351B (en) | Method for preparing polyvinylidene fluoride (PVDF)/barium titanate (BT) composite membrane materials | |
| CN103839689B (en) | Electrode of used by dye sensitization solar battery dopen Nano gold and preparation method thereof | |
| CN105271214B (en) | A kind of redox preparation method of graphene | |
| CN111697149A (en) | Sulfate modified perovskite thin film, preparation method and application thereof | |
| CN105036108B (en) | A kind of method that utilization maize leaves prepare sodium-ion battery electrode material with carbon element | |
| CN107436314A (en) | One kind is based on TiO2The moisture sensor of composite | |
| CN104726851B (en) | A kind of method that sol-gal process prepares p-type SnO 2 thin film material | |
| CN110395723A (en) | A kind of method of fuse salt strategy preparation High-performance graphene | |
| CN109301069A (en) | Solar cell and preparation method thereof | |
| Ibrahim et al. | The impact of annealing process on the grain morphology and performance of mesoporous nip carbon-based perovskite solar cells | |
| CN109950397A (en) | A kind of preparation process of high performance calcium titanium ore solar battery thin film | |
| CN112981484B (en) | Method for preparing perovskite material based on electrochemical method | |
| CN109666482A (en) | A kind of preparation method of rare-earth-doped fluoride up-conversion luminescence nano-powder |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160907 |