CN110055307B - Ultra-trace multi-element biomacromolecule detection method - Google Patents
Ultra-trace multi-element biomacromolecule detection method Download PDFInfo
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- CN110055307B CN110055307B CN201910327126.2A CN201910327126A CN110055307B CN 110055307 B CN110055307 B CN 110055307B CN 201910327126 A CN201910327126 A CN 201910327126A CN 110055307 B CN110055307 B CN 110055307B
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C12Q1/6813—Hybridisation assays
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Abstract
The invention discloses a method for detecting ultra-trace multi-element biomacromolecules based on a mixed hydrogel magnetic photonic crystal bar code, wherein the mixed hydrogel magnetic photonic crystal bar code is obtained by taking a protein structure formed by self-assembling monodisperse microspheres prepared by a microfluidic device as a template, then pouring mixed magnetic nanoparticle hydrogel into the template and removing the template; and then coupling a probe on the surface of the mixed hydrogel photonic crystal bar code, integrating the probe on a super-hydrophobic surface, and naturally concentrating and enriching the sample by utilizing the wettability difference so as to achieve the purpose of detecting the ultra-trace biomacromolecules. Based on stable optical characteristics such as characteristic reflection peaks of photonic crystals, the detection of various biomacromolecules, namely multi-element detection, can be realized at the same time. In conclusion, the concentration and enrichment method provided by the invention is simple to operate, low in cost, high in sensitivity and high in universality, and can be applied to clinical ultrasensitive detection of multielement micro biomacromolecules.
Description
Technical Field
The invention relates to a biomacromolecule detection method, in particular to an ultra-trace multielement biomacromolecule detection method based on a mixed hydrogel magnetic photonic crystal bar code and driven to concentrate by natural evaporation.
Background
Biological macromolecules (such as nucleic acids, proteins, etc.) are important components of the human body. Identification of human biological macromolecules is an important strategy for clinical diagnosis, DNA research, bacterial/viral infection, drug development, mutation analysis, and gene therapy. However, the biomacromolecules existing in human blood are small in size and high in dilution degree, and accuracy and sensitivity in the biomacromolecule detection process are seriously influenced. How to realize ultrasensitive multiplex detection of biomacromolecules at low density is an important obstacle for restricting the development of detection technology.
Currently, there are several signal amplification techniques for detecting biomacromolecules. The most common methods of nucleic acid amplification are Polymerase Chain Reaction (PCR), rolling Circle Amplification (RCA), strand Displacement Amplification (SDA), and Hybrid Chain Reaction (HCR). However, PCR is prone to non-specific amplification, false negatives, and false positives. The background problem of signal detection and the cost of the lock probe limit the development of RCA. SDA and HCR cannot achieve great economic benefit due to high cost and long time consumption. Enzyme-linked immunoassay, radioimmunoassay, fluorescence immunoassay and the like are the most commonly used protein determination methods at present, but the methods still have the problems of complicated operation, low sensitivity and the like. Therefore, a biomacromolecule detection method which is simple and convenient to operate, low in detection cost and high in sensitivity needs to be developed. In many strategies, the target is enriched to a sensitive area from a highly diluted solution, and a stronger signal and a lower detection limit are obtained by increasing the concentration of the sensitive area, so that the method is an important scheme for realizing the ultrasensitive multiple detection of biomacromolecules at low density.
Inspired by the bionic enrichment of super-wettability materials, the invention concentrates and enriches highly diluted solution to a hydrophilic sensitive area through the surface wettability difference. In addition, because the characteristic reflection peak generated by the photonic band of the photonic crystal depends on the physical structure of the photonic band sequence arrangement, the method constructs a coding signal which has good stability and is not interfered by any fluorescence background and photobleaching based on the photonic crystal, realizes visual and high-sensitivity multi-element detection, and effectively improves the detection accuracy and sensitivity of different targets such as biomacromolecules.
Disclosure of Invention
The purpose of the invention is as follows: the invention provides a method for detecting ultra-trace multi-element biomacromolecules, which aims at solving the problems of inconvenient operation, high cost, difficult sample recovery and the like of the prior art, and realizes the concentration of a detection target by utilizing the surface wettability difference and natural evaporation drive concentration so as to achieve the effect of ultra-sensitive detection.
The technical scheme is as follows: the invention relates to an ultra-trace multielement biomacromolecule detection method, which integrates a mixed hydrogel photonic crystal bar code containing magnetic nano particles on a super-hydrophobic surface for detection and specifically comprises the following steps:
(1) Preparing a mixed hydrogel photonic crystal bar code containing magnetic nanoparticles:
(2) Preparing a super-hydrophobic surface:
(3) Ultrasensitive detection of biological macromolecules: carrying out group activation on the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles prepared in the step (1), modifying a probe or an antibody on the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles after group coupling, then integrating the modified probe or antibody on the super-hydrophobic surface prepared in the step (2), and carrying out nucleic acid detection by dropwise adding a target biomacromolecule sample to be detected;
(4) Magnetic control washing is carried out, so that the accuracy of subsequent detection is improved.
Wherein, step (1) includes the following steps: based on a capillary assembly microfluidic chip, preparing single emulsion with controllable particle size by controlling the flow of the discrete phase and the continuous phase, and cleaning and collecting the single emulsion; then, self-assembling the single emulsion into an opal structure template by using a chemical vapor deposition method, and then pouring the hydrogel mixed with the magnetic nanoparticles into the opal structure template; and finally, polymerizing the hydrogel under ultraviolet illumination, and removing the template by using a remover to obtain the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles.
The discrete phase is selected from one or more than two materials of silicon dioxide nano-particle aqueous solution, polystyrene, benzene and dichloroethane; the continuous phase is selected from methyl silicone oil and polyvinyl alcohol.
The particle size of the magnetic nano-particles is between 5 and 50 nm; the hydrogel is selected from polyethylene glycol diacrylate, polyisopropylacrylamide, acrylamide and acrylic acid.
The remover is one of hydrofluoric acid, sodium hydroxide or acetone.
The particle size of the single emulsion is between 200 and 300 mu m.
Further, the step (2) comprises the following steps: and uniformly dispersing the micron particles on the surface of a clean glass slide, preparing a particle film on the surface of the glass slide through self-assembly, and performing super-hydrophobic treatment on the particle film to obtain the super-hydrophobic surface.
The micro-particles are one or two of silica particles and polystyrene particles.
The super-hydrophobic treatment is to adopt 1-5% of trimethoxy silane dichloromethane solution in volume ratio to prepare a super-hydrophobic substrate by evaporation for 2-5 hours.
In the step (4), the mixed hydrogel magnetic photonic crystal bar code after the detection reaction in the step (3) is fixed by a magnet, and redundant probes or antibodies and fluorescent markers thereof are washed away by phosphate buffer solution, so that the accuracy of subsequent detection is facilitated. Typically 3-4 washes.
Has the advantages that: compared with other traditional enrichment methods, the method for detecting the ultra-trace multi-element biomacromolecules based on the mixed hydrogel magnetic photonic crystal bar code integrates the mixed hydrogel magnetic photonic crystal bar code coupled with a probe or an antibody on a super-hydrophobic surface, and enriches and concentrates a sample on the photonic crystal bar code by using wettability difference and natural evaporation so as to realize ultra-sensitive detection. The invention can realize the concentration rapid enrichment of the target detection object, improves the detection sensitivity, has simple preparation and low cost for detecting the photonic crystal bar code, has high universality due to the magnetic characteristic, and greatly enhances the detection of ultra-trace multi-element biomacromolecules.
Drawings
FIG. 1 is a schematic diagram of the process for preparing the hybrid hydrogel magnetic photonic crystal barcode of the present application;
FIG. 2 is a schematic diagram of the application of the hybrid hydrogel magnetic photonic crystal barcode to the detection of ultrasensitive biomacromolecules;
FIG. 3 is a schematic view of a washing process of a magnetic controlled photonic crystal barcode of the present application;
FIG. 4 shows the detection result of the magnetic control photonic crystal barcode for detecting nucleic acid.
Detailed Description
The present application will now be described in detail with reference to the drawings and specific examples.
The invention discloses a method for detecting ultra-trace multi-element biomacromolecules, which comprises the following steps:
(1) Preparing a mixed hydrogel photonic crystal bar code containing magnetic nanoparticles:
(2) Preparing a super-hydrophobic surface:
(3) Ultrasensitive detection of biological macromolecules:
(4) And (5) washing by magnetic control.
Wherein, figure 1 shows a schematic diagram of the preparation process of the hybrid hydrogel magnetic photonic crystal bar code. Firstly, preparing a microsphere template by utilizing microfluidics, then forming an inverse opal structure by utilizing self-assembly, then filling hydrogel with magnetic nanoparticles into the microsphere, and finally removing the template to obtain the magnetic hydrogel photonic crystal. FIG. 2 shows a schematic diagram of the application of the mixed hydrogel magnetic photonic crystal bar code to the detection of ultrasensitive biomacromolecules. Integrating the magnetic photonic crystal bar code coupled with the probe or the antibody on a super-hydrophobic surface, then dripping a target sample to be detected into a photonic crystal bar code area, and concentrating and enriching a target substance to the photonic crystal bar code along with natural evaporation so as to realize super-sensitive detection. FIG. 3 shows the washing process of magnetically controlled photonic crystal bar code. Fixing the mixed hydrogel magnetic photonic crystal bar code after the detection reaction in the third step by a magnet, washing the mixed hydrogel magnetic photonic crystal bar code for a plurality of times by using a phosphate buffer solution to wash off redundant probes or antibodies and fluorescent markers thereof, so as to facilitate the accuracy of subsequent detection.
Reference is made to the preparation of microfluidic chips described in the present application: nanoscale,2013,5,9553.
Example 1
An ultra-trace multi-element biomacromolecule detection method comprises the following steps:
(1) Preparation of silica single emulsion:
the discrete phase for preparing the single emulsion is a silicon dioxide aqueous solution, and the mass volume ratio is 20%. The continuous phase is methyl silicone oil, and the viscosity is 50cs. The flow rate of the discrete phase was 0.5mL/h, and the flow rate of the continuous phase was 3mL/h. In the micro-fluidic chip, the inertia force and the viscous force act together to shear the dispersed phase into single emulsion. The single emulsion was then collected into a methyl silicone oil collection tank containing 500 cs.
(2) Single emulsion solidification:
and (3) transferring the collecting tank filled with the single emulsion into an oven with the temperature of 80 ℃, standing for 12 hours to volatilize part of water in the emulsion, soaking the emulsion in normal hexane for 20 minutes in the collecting tank to remove silicone oil, and then putting the microspheres into a muffle furnace to calcine and solidify the microspheres at the temperature of 800 ℃.
(3) Copying and removing the template microspheres to obtain a magnetic mixed hydrogel photonic crystal bar code:
and (3) placing the silica microspheres at the bottom of a centrifuge tube, adding pure water, and drying in an oven at 60 ℃ to obtain the regularly arranged silica template at the bottom of the centrifuge tube. Mixing 10% of polyethylene glycol diacrylate (PEGDA) and 10% of Acrylic Acid (AA) in a volume ratio of 1:1 mix and add 10% magnetic nanoparticles (4 mg/ml) and 1% photoinitiator. And adding the mixed solution into a silicon dioxide template, irradiating the polymerized hydrogel by using ultraviolet light after 30 minutes, and finally removing silicon dioxide by using hydrofluoric acid to obtain the bar code.
(4) Preparing a super-hydrophobic surface:
uniformly dispersing a silicon dioxide alcohol solution with the particle size of 4 mu m on the surface of the glass slide, self-assembling silicon dioxide into a silicon dioxide photonic crystal film, and then evaporating and plating 1% trimethoxy silane dichloromethane solution for 2 hours to prepare the super-hydrophobic photonic crystal film;
(5) And (3) biological macromolecule detection:
and (2) putting the prepared mixed hydrogel photonic crystal bar code into 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to be dissolved in morpholine ethanesulfonic acid (MES) buffer solution for 1 hour of carboxyl activation, adding an amino modified nucleic acid probe for probe coupling, washing off redundant probes, integrating the bar code on a super-hydrophobic surface, dropwise adding a nucleic acid sample to be detected into a bar code area, and naturally evaporating, concentrating and hybridizing the nucleic acid sample. The magnetic bar code is fixed by a magnet, washed three times by Phosphate Buffer Solution (PBS), and finally the detection result is judged by the fluorescence signal of the fluorescence marker. The detection result is shown in figure 4, and the graph shows that the fluorescence intensity is linearly related to the logarithm of the concentration of the nucleic acid sample, and the magnitude order of the detection limit is 10 -14 Compared with the traditional detection method, the method greatly improves the detection sensitivity.
Example 2
An ultra-trace multi-element biomacromolecule detection method comprises the following steps:
(1) Preparation of monodisperse polystyrene emulsion:
the discrete phase for preparing the single emulsion was Polystyrene (PS)/benzene/dichloroethane, the mass to volume ratio was 0.5g, 10ml, the external phase was an aqueous solution of polyvinyl alcohol (PVA), the mass to volume ratio was 5%. The oil phase of PS is cut into oil-in-water single emulsion structures with consistent diameters by the water phase of PVA through a stable liquid flow rate (inward flow rate is 0.5 mL/h) and a shearing force (outward flow rate is 5 mL/h), and then the emulsion enters a collecting pool filled with the water phase of PVA through a collecting pipe. As a surfactant, the existence of PVA can effectively prevent the mutual fusion phenomenon of the emulsions in the collection process;
(2) Curing the polystyrene emulsion:
transferring the prepared emulsion drop to a flask of a rotary evaporator, increasing the temperature by 5 ℃ every 5 minutes from room temperature to 60 ℃, and controlling the rotating speed to be 40rpm; after the temperature reaches 60 ℃, the temperature rising speed is reduced to 2 ℃ every 30 minutes, and the temperature rising is stopped when the temperature reaches 70 ℃; after the temperature exceeds 64 ℃, the rotating speed is increased to 80rpm; after stopping the temperature rise, 1 hour, the curing was checked. Repeatedly cleaning the cured PS microspheres with ultrapure water, and drying in a 60 ℃ oven;
(3) Copying and removing the template microspheres to obtain a magnetic mixed hydrogel photonic crystal bar code:
mixing 10% of polyethylene glycol diacrylate (PEGDA) and 10% of Acrylic Acid (AA) in a volume ratio of 1:1 mix and add 10% magnetic nanoparticles (4 mg/ml) and 1% photoinitiator. And adding the mixed solution into a PS template, irradiating the polymerized hydrogel by using ultraviolet light after 30 minutes, and finally removing polystyrene by using acetone to obtain the bar code.
(4) Preparing a super-hydrophobic surface:
uniformly dispersing a silicon dioxide alcohol solution with the particle size of 4 mu m on the surface of the glass slide, self-assembling silicon dioxide into a silicon dioxide photonic crystal film, and then evaporating and plating a trimethoxy silane dichloromethane solution for 2 hours to prepare a super-hydrophobic photonic crystal film;
(5) And (3) biological macromolecule detection:
and (3) putting the prepared mixed hydrogel photonic crystal bar code into an Epichlorohydrin (ECH) and sodium hydroxide (NaOH) solution for activating an epoxy group for 1 hour, adding an antibody for antibody coupling, washing off redundant antibody, integrating the bar code on a super-hydrophobic surface, dripping a protein sample to be detected into a bar code area, and naturally evaporating, concentrating and hybridizing the protein sample. The magnetic bar code is fixed by a magnet, washed three times by Phosphate Buffer Solution (PBS), and finally the detection result is judged by the fluorescence signal of the fluorescence marker.
Claims (3)
1. A method for detecting an ultra-trace multielement biological macromolecule is characterized in that a mixed hydrogel photonic crystal bar code containing magnetic nanoparticles is integrated on a super-hydrophobic surface for detection, and comprises the following steps:
(1) Preparing a mixed hydrogel photonic crystal bar code containing magnetic nanoparticles: based on a capillary assembly microfluidic chip, preparing single emulsion with controllable particle size by controlling the flow of the discrete phase and the continuous phase, and cleaning and collecting the single emulsion; then, self-assembling the single emulsion into an opal structure template by using a chemical vapor deposition method, and then pouring the hydrogel mixed with the magnetic nanoparticles into the opal structure template; finally, polymerizing hydrogel under ultraviolet illumination, and removing the template by adopting a remover to obtain the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles; the particle size of the single emulsion is 200-300 mu m; the particle size of the magnetic nano-particles is 5-50nm, and the remover is hydrofluoric acid, sodium hydroxide or acetone;
(2) Preparing a super-hydrophobic surface: uniformly dispersing micron particles on the surface of a clean glass slide, preparing a particle film on the surface of the glass slide through self-assembly, and performing super-hydrophobic treatment on the particle film to obtain a super-hydrophobic surface; the super-hydrophobic treatment is to adopt a trimethoxy silane dichloromethane solution with the volume ratio of 1 to 5% to prepare a super-hydrophobic substrate by evaporation for 2 to 5 hours; the micrometer particles are one or two of silicon dioxide particles and polystyrene particles;
(3) Ultrasensitive detection of biological macromolecules: carrying out group activation on the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles prepared in the step (1), modifying a probe or an antibody on the mixed hydrogel photonic crystal bar code containing the magnetic nanoparticles after group coupling, then integrating the modified probe or antibody on the super-hydrophobic surface prepared in the step (2), and carrying out nucleic acid detection by dropwise adding a target biomacromolecule sample to be detected;
(4) And (4) performing magnetic control washing, fixing the mixed hydrogel magnetic photonic crystal bar code after the detection reaction in the step (3) by a magnet, and washing redundant probes or antibodies and fluorescent markers thereof by using a phosphate buffer solution so as to facilitate the accuracy of subsequent detection.
2. The method for detecting the ultra-trace multi-element biological macromolecules of claim 1, wherein the discrete phase is one or more than two materials selected from a silica nanoparticle aqueous solution, polystyrene, benzene and dichloroethane; the continuous phase is selected from methyl silicone oil and polyvinyl alcohol.
3. The method for detecting the ultra-trace multi-element biomacromolecule as claimed in claim 1, wherein the hydrogel is selected from polyethylene glycol diacrylate, polyisopropylacrylamide, acrylamide and acrylic acid.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106040114A (en) * | 2016-05-24 | 2016-10-26 | 华中科技大学 | Hydrogel photonic crystal microspheres, and preparation method and application thereof |
| CN106566528A (en) * | 2016-11-07 | 2017-04-19 | 合肥亨纳生物科技有限公司 | Up-conversion fluorescence microsphere capable of being coded and preparation method thereof |
| CN108373518A (en) * | 2018-05-14 | 2018-08-07 | 东南大学 | A kind of mixing photonic crystal and the preparation method and application thereof |
| CN108546682A (en) * | 2018-05-14 | 2018-09-18 | 东南大学 | A kind of mixing photon crystal composite material and application based on modified with folic acid |
| CN108641025A (en) * | 2018-05-14 | 2018-10-12 | 东南大学 | A method of preparing the mixing photon crystal composite material based on modified with folic acid |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011034678A1 (en) * | 2009-09-21 | 2011-03-24 | Arizona Board Of Regents, A Body Corporate Of The State Of Arizona | Use of superhydrophobic surfaces for liquid agglutination assays |
-
2019
- 2019-04-23 CN CN201910327126.2A patent/CN110055307B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106040114A (en) * | 2016-05-24 | 2016-10-26 | 华中科技大学 | Hydrogel photonic crystal microspheres, and preparation method and application thereof |
| CN106566528A (en) * | 2016-11-07 | 2017-04-19 | 合肥亨纳生物科技有限公司 | Up-conversion fluorescence microsphere capable of being coded and preparation method thereof |
| CN108373518A (en) * | 2018-05-14 | 2018-08-07 | 东南大学 | A kind of mixing photonic crystal and the preparation method and application thereof |
| CN108546682A (en) * | 2018-05-14 | 2018-09-18 | 东南大学 | A kind of mixing photon crystal composite material and application based on modified with folic acid |
| CN108641025A (en) * | 2018-05-14 | 2018-10-12 | 东南大学 | A method of preparing the mixing photon crystal composite material based on modified with folic acid |
Non-Patent Citations (4)
| Title |
|---|
| Colour-barcoded magnetic microparticles for multiplexed bioassays;Howon Lee1 等;《NATURE MATERIALS》;20100930;第9卷;第745-749页 * |
| Emerging barcode particles for multiplex bioassays;Yueshuang Xu等;《SCIENCE CHINA Materials》;20180910;第62卷(第3期);第289–324页 * |
| 光子晶体传感器研究进展;侯觉等;《中国科学:化学》;20161020(第10期);摘要 * |
| 胶体光子晶体膜的超浸润性研究进展;崔丽影等;《化学学报》;20171015(第10期);摘要，第969页左栏第2段，第974页右栏第1段至第975页左栏第1-2段 * |
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