CN111443049A - Preparation method and application of colorimetric array sensor of zirconium-based metal-organic framework and single-stranded DNA modified gold nanoparticles - Google Patents
Preparation method and application of colorimetric array sensor of zirconium-based metal-organic framework and single-stranded DNA modified gold nanoparticles Download PDFInfo
- Publication number
- CN111443049A CN111443049A CN201910046219.8A CN201910046219A CN111443049A CN 111443049 A CN111443049 A CN 111443049A CN 201910046219 A CN201910046219 A CN 201910046219A CN 111443049 A CN111443049 A CN 111443049A
- Authority
- CN
- China
- Prior art keywords
- zirconium
- stranded dna
- gold nanoparticles
- organic framework
- based metal
- 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.)
- Granted
Links
- 108020004414 DNA Proteins 0.000 title claims abstract description 53
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 51
- 102000053602 DNA Human genes 0.000 title claims abstract description 45
- 108020004682 Single-Stranded DNA Proteins 0.000 title claims abstract description 45
- 239000013096 zirconium-based metal-organic framework Substances 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 150000002343 gold Chemical class 0.000 claims abstract description 33
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 31
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 31
- 210000000582 semen Anatomy 0.000 claims abstract description 25
- 230000003993 interaction Effects 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 16
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000010931 gold Substances 0.000 claims abstract description 15
- 229910052737 gold Inorganic materials 0.000 claims abstract description 15
- 238000004458 analytical method Methods 0.000 claims abstract description 12
- 239000002244 precipitate Substances 0.000 claims abstract description 8
- 208000007466 Male Infertility Diseases 0.000 claims abstract description 7
- 230000004044 response Effects 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 19
- QCWXUUIWCKQGHC-UHFFFAOYSA-N Zirconium Chemical compound [Zr] QCWXUUIWCKQGHC-UHFFFAOYSA-N 0.000 claims description 13
- 238000002835 absorbance Methods 0.000 claims description 13
- 239000002243 precursor Substances 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 230000008859 change Effects 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000011534 incubation Methods 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 6
- 239000001509 sodium citrate Substances 0.000 claims description 6
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 6
- 229940038773 trisodium citrate Drugs 0.000 claims description 6
- 239000013110 organic ligand Substances 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 239000004809 Teflon Substances 0.000 claims description 4
- 229920006362 Teflon® Polymers 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 4
- BSDOQSMQCZQLDV-UHFFFAOYSA-N butan-1-olate;zirconium(4+) Chemical compound [Zr+4].CCCC[O-].CCCC[O-].CCCC[O-].CCCC[O-] BSDOQSMQCZQLDV-UHFFFAOYSA-N 0.000 claims description 4
- VDNSHGNZYOIMOW-UHFFFAOYSA-N oxygen(2-) zirconium(4+) Chemical compound [O--].[O--].[O--].[O--].[Zr+4].[Zr+4] VDNSHGNZYOIMOW-UHFFFAOYSA-N 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- XPGAWFIWCWKDDL-UHFFFAOYSA-N propan-1-olate;zirconium(4+) Chemical compound [Zr+4].CCC[O-].CCC[O-].CCC[O-].CCC[O-] XPGAWFIWCWKDDL-UHFFFAOYSA-N 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- VPGLGRNSAYHXPY-UHFFFAOYSA-L zirconium(2+);dichloride Chemical compound Cl[Zr]Cl VPGLGRNSAYHXPY-UHFFFAOYSA-L 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 3
- 230000008836 DNA modification Effects 0.000 claims description 2
- 239000013207 UiO-66 Substances 0.000 claims description 2
- 238000003556 assay Methods 0.000 claims description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 230000002194 synthesizing effect Effects 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 230000027455 binding Effects 0.000 abstract description 4
- 230000002776 aggregation Effects 0.000 abstract description 3
- 238000004220 aggregation Methods 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract 1
- 239000012621 metal-organic framework Substances 0.000 description 22
- GBNDTYKAOXLLID-UHFFFAOYSA-N zirconium(4+) ion Chemical compound [Zr+4] GBNDTYKAOXLLID-UHFFFAOYSA-N 0.000 description 20
- 239000000523 sample Substances 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 125000000524 functional group Chemical group 0.000 description 4
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- GPNNOCMCNFXRAO-UHFFFAOYSA-N 2-aminoterephthalic acid Chemical compound NC1=CC(C(O)=O)=CC=C1C(O)=O GPNNOCMCNFXRAO-UHFFFAOYSA-N 0.000 description 2
- 239000005711 Benzoic acid Substances 0.000 description 2
- 229910007932 ZrCl4 Inorganic materials 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 235000010233 benzoic acid Nutrition 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000009881 electrostatic interaction Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000527 sonication Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- DUNKXUFBGCUVQW-UHFFFAOYSA-J zirconium tetrachloride Chemical compound Cl[Zr](Cl)(Cl)Cl DUNKXUFBGCUVQW-UHFFFAOYSA-J 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 206010021929 Infertility male Diseases 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000009612 semen analysis Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 230000019100 sperm motility Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a preparation method and application of a colorimetric array sensor of zirconium-based metal-organic framework and single-stranded DNA modified gold nanoparticles. 6 different single-stranded DNA sequences are designed to be modified on the surface of the gold nanoparticle to form a unique sensor recognition element. The novel material zirconium-based metal organic framework can combine and precipitate single-stranded DNA sequences to modify gold nanoparticles through different intermolecular forces, and after certain protein or semen samples are added, the binding force of the protein or semen samples can be accelerated or slowed down, so that the aggregation behavior of the protein or semen samples is influenced, and a detectable signal is generated. Thus, we can consider these recognition elements to have different interactions with different target binding, and we can clearly detect different targets by analyzing the colorimetric response of each recognition element in combination. The invention can be simply and efficiently used for colorimetric array analysis of various proteins and male semen and provides a detection basis for analysis of male infertility.
Description
Technical Field
The invention belongs to the field of analytical chemistry, and particularly relates to a preparation method and application of a colorimetric sensor of zirconium ion metal-organic framework and single-stranded DNA modified gold nanoparticles
Background
Male sterility is a social disease that afflicts male siblings, and the rapid and accurate identification of sperm motility is the key to diagnosis and formulation of targeted treatment strategies. In addition to known factors, there are still 30-45% of patients whose causes of abnormalities in the parameters of the semen have not been identified, and the cases of these abnormalities are collectively called idiopathic male infertility. The threshold for the normal range has not been determined unambiguously due to the lack of available data on the most direct sample of semen variations. Furthermore, because of the complexity and diversity of the sample, many limitations are imposed on the detection. The detection of proteomes in semen plays an important role in all levels. However, many detection systems are not capable of performing multiple assays simultaneously due to the limitations of specific binding interactions between the protein and the receptor. Given the growing demand for medical diagnostics, a variety of protein analysis methods that are fast, simple, and sensitive, rather than "key-locked" specific identification strategies, are urgently needed.
The method allows for the generation of unique fingerprints for each analyte in a manner similar to the mammalian olfactory system, presenting chemical diversity, and these unique responses can be analyzed by linear discriminant analysis (L DA), providing a multifunctional system that can not only detect individual but also mixed targets6The coordination interaction between the clusters and the DNA phosphate backbone successfully binds to the oligonucleotide. And the MOFs structure contains aromatic electron-rich ligands which can be combined with single-stranded DNA through pi-pi stacking and H-bonds. These superior properties suggest that MOFs may have great potential for application to array sensing for semen analysis.
Disclosure of Invention
The invention aims to provide a colorimetric array sensor based on interaction of a zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles, which is used for detection and analysis of various proteins and male semen.
The invention mainly combines the design of 6 different single-stranded DNA sequences to be modified on the surface of gold nanoparticles to form a unique sensor identification element. The zirconium ion metal organic framework and the single-stranded DNA modified gold nanoparticles are rich in various functional groups, and different intermolecular forces such as hydrogen bond attraction, electrostatic interaction and other chemical bonds for interacting with target proteins exist among the functional groups. The zirconium ion metal organic framework can precipitate single-stranded DNA modified gold nanoparticles, and after some proteins are added, the binding force of the proteins can be accelerated or slowed down, the aggregation behavior of the proteins is influenced, and therefore a detectable signal is generated. Thus, we can consider these recognition elements to have different interactions with different target binding, and we can clearly detect different targets by analyzing the colorimetric response of each recognition element in combination. The invention can be simply and efficiently used for colorimetric array analysis of various proteins and male semen and provides a detection basis for analysis of male infertility. The signal detection of the colorimetric array sensor does not need expensive instruments and equipment, the cost of the preparation process is low, the operation is simple in the using process, and the colorimetric array sensor is suitable for instant detection.
The technical scheme of the invention comprises the following steps:
a colorimetric array sensor based on interaction of a zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles is characterized by comprising a zirconium ion metal organic framework and the single-stranded DNA modified gold nanoparticles.
Wherein, the zirconium ion metal organic framework has a formula conforming to one of UiO-66(Zr) -X substances, and X can be one of-NH 2, -NO2, - (OH)2, -Br and-COOH.
Wherein the size of the zirconium ion metal organic framework is 70-500 nm.
Wherein the size of the gold nanoparticles is 10-200 nm.
Wherein the single-stranded DNA comprises a random combination of thiol-modified PolyA, Poly T, Poly C, and A, T, C, G.
Wherein the length of the single-stranded DNA is 5-50 bp.
The colorimetric array sensor with the interaction of the zirconium-based metal organic framework and the single-stranded DNA modified gold nanoparticles comprises the following steps:
(1) the preparation method of the zirconium ion metal organic framework comprises the steps of dissolving a zirconium metal precursor and an organic ligand in 10m L DMF, transferring the zirconium metal precursor and the organic ligand into a Teflon lining, carrying out ultrasonic treatment, and carrying out high-temperature standing reaction, wherein the zirconium metal precursor of the zirconium metal precursor comprises at least one of zirconium tetraoxide, zirconium dichloride, zirconium n-butoxide and zirconium n-propoxide, the concentration of the zirconium metal precursor is 5-25mg/m L, the zirconium metal precursor of the organic ligand comprises at least one of zirconium tetraoxide, zirconium dichloride, zirconium n-butoxide and zirconium n-propoxide, the concentration of the zirconium metal precursor is 5-20mg/m L, the reaction temperature is 100-200 ℃, and the reaction time is 10-24 hours.
(2) The preparation of the gold nanoparticles comprises the steps of mixing 50m L chloroauric acid and 5m L trisodium citrate, heating, reacting and synthesizing the gold nanoparticles, wherein the concentration of the chloroauric acid is 1-5mM, the concentration of the trisodium citrate is 20-50mM, and the reaction time is 10-60 minutes.
(3) The gold nanoparticles are subjected to single-stranded DNA modification: thiolated DNA strands are activated by reaction with tris (2-carboxyethyl) phosphine and incubated with the prepared colloidal solution of AuNPs. Thereafter, 0.1M NaCl was gradually added to the mixture to "age" and incubation was continued. The single-stranded DNA comprises a random combination of thiol-modified PolyA, Poly T, Poly C, and A, T, C, G; wherein, the length of the single-stranded DNA is 5-50 bp; wherein the concentration of the tri (2-carboxyethyl) phosphine is 1-50 mM; wherein the incubation time is 5-24 hours and the incubation temperature is 25-40 ℃.
(4) And (4) detecting the protein sample or the semen.
The using method comprises the following steps:
(1) zirconium ion metal organic frameworks and single stranded DNA modified gold nanoparticles were diluted with Tris-HCl (PH 7-7.8) to the final appropriate concentration.
(2) In a 96-well microplate, a certain amount of different single-stranded DNA-modified gold nanoparticles were separately transferred to each well, and the absorption intensity of the solution was recorded at 520nm by a Safire microplate reader (A)0)
(3) Adding different analytes with certain concentration, multiple proteins or different semen samples, and incubating for 30-120 min at 25-40 deg.C.
(4) Adding a certain amount of zirconium ion metal organic framework into the mixture solution, and reacting for 2-12 hours at the temperature of 25-40 ℃ to combine and precipitate the residual single-stranded DNA modified gold nanoparticles or proteins.
(5) The supernatant without precipitation was carefully transferred to a new 96-well microplate and the absorbance intensity (A), the relative absorbance intensity change (A) was measured0-A) is used as the absorbance response change value.
(6) And (4) carrying out linear discriminant analysis on the original data matrix by using corresponding software.
The colorimetric array sensor based on the interaction of the zirconium-based metal organic framework and the single-stranded DNA modified gold nanoparticles can be used for analyzing and detecting a plurality of proteins at low concentration.
The colorimetric array sensor with the interaction of the zirconium-based metal organic framework and the single-stranded DNA modified gold nanoparticles can be used for analyzing and detecting semen of different types of male infertility.
The principle of the color change of the invention is as follows:
the principle is shown in figure 1, zirconium ion metal organic framework and single-stranded DNA modified gold nanoparticles are rich in various functional groups, and different intermolecular forces such as hydrogen bond attraction, electrostatic interaction and other chemical bonds for interacting with target proteins exist among the functional groups. The zirconium ion metal organic framework can precipitate single-stranded DNA modified gold nanoparticles. After a sample to be detected is added, such as a male semen sample, some proteins, enzymes, and other contents contained in semen can be adsorbed to the surface of the single-stranded DNA modified gold nanoparticles, then the combination of the single-stranded DNA modified gold nanoparticles and the zirconium ion metal organic framework is prevented, and after the zirconium ion metal organic framework is introduced, only the remaining single-stranded DNA modified gold nanoparticles can be precipitated by the zirconium ion metal organic framework, so that the aggregation efficiency and the color of the supernatant are changed. Differential interactions between different contents and single stranded DNA modified gold nanoparticles or zirconium ion metal organic frameworks will produce different colorimetric modes. In a sensing study, we were able to easily distinguish between different semen samples by recording the value of the absorbance signal of the supernatant before and after addition of protein.
The colorimetric array sensor based on the interaction of the zirconium-based metal organic framework and the single-stranded DNA modified gold nanoparticles has at least the following beneficial effects:
(1) the invention uses the interaction of the zirconium ion metal organic framework and the single-stranded DNA modified gold nanoparticles for the first time, and is applied to the detection of biological samples.
(2) The colorimetric array sensor prepared by the invention not only can realize the detection of various proteins at the same time, but also can analyze different semen samples.
(3) The colorimetric array sensor prepared by the invention has the advantages of low manufacturing cost, simple process, easy operation and stable reaction system, and is not influenced by samples.
(4) The invention is also expected to be used for the diagnosis of other diseases, the analysis and the prognosis diagnosis of tumor cells, provides a new idea for the clinical detection of diseases and has wide application prospect.
Drawings
FIG. 1 is a schematic diagram of the present invention
FIG. 2 is an L DA profile of the detection of various proteins
FIG. 3 is a L DA analysis of different semen samples
Detailed Description
Example 1: detection of multiple proteins by colorimetric array sensor with interaction of zirconium ion metal organic framework and single-stranded DNA modified gold nanoparticles
The method comprises the following steps: preparation of zirconium ion metal organic framework: reacting ZrCl4(120mg), aminoterephthalic acid (110mg) and benzoic acid (1.9g) were dissolved in 10m L DMF and transferred to a Teflon liner after sonication for 3 minutes, placed at 120 ℃ for standing reaction for 24 hours the resulting product was obtained by centrifugation and washed repeatedly three times with fresh DMF and methanol (8,000rpm, 5 minutes). Finally, to remove residual organic solvent, the nanoparticles were collected after drying overnight at 65 ℃.
Step one, gold nanoparticle preparation, namely, when a chloroauric acid solution (50m L, 1mM) is heated to boiling, a trisodium citrate solution (5m L, 38.8mM) is rapidly added under vigorous stirring, the mixed solution is continuously heated and fully stirred for 30 minutes to ensure that the reaction is finished, and after cooling, the dark red solution is stored at 4 ℃ for use.
Step three: modification of single-stranded DNA by gold nanoparticles
6 different DNA strands (NP 1: SH- (CH)2)6-TTTTT；NP2：SH-(CH2)6-TTTTTTTTTTTTTTT； NP3：SH-(CH2)6-AAAAAAAAAAAAAAA；NP4：SH-(CH2)6-CCCCC；NP5：SH-(CH2)6-C CCCCCCCCCCCCCC；NP6：SH-(CH2)6-GAGAGAGAGAGAGAG) was activated with Tris (2-carboxyethyl) phosphine (TC EP) (10mM) for 1 hour, then incubated with the prepared AuNPs colloidal solution for 16 hours (final DNA concentration of 3 μ M.) after which 0.1M NaCl was gradually added to the mixture to "age" and incubation was continued for 24 hours.
Step four: detection of the protein: ssDNA-AuNPs (NP1-NP6) and Zr-MOF were diluted with Tris-HCl (pH 7-7.8) to the final appropriate concentration. In a 96-well microplate, a certain amount of different ssDNA-AuNPs were transferred to each well, respectively, and the absorption intensity of the solution was recorded at 520nm by means of a Safire microplate reader (A)0) Adding different protein dilutions with certain concentrations, and incubating at 25-40 deg.C for 30-120 min. (4) Adding a certain amount of Zr-MOF into the mixture solution, and reacting for 2-12 hours in an environment of 25-40 ℃ to combine and precipitate the residual ssDNA-AuNP or protein. The supernatant without precipitation was carefully transferred to a new 96-well microplate and the absorbance intensity (A), the relative absorbance intensity change (A) was measured0-A) is used as the absorbance response change value. Soft with SPSS v25.0Results are shown in fig. 2, L DA can identify different proteins by very subtle differences, which demonstrates the effectiveness of our method.
Example 2: detection of various semen samples by colorimetric array sensor with interaction of zirconium ion metal organic framework and single-stranded DNA modified gold nanoparticles
The method comprises the following steps: preparation of zirconium ion metal organic framework: reacting ZrCl4(120mg), aminoterephthalic acid (110mg) and benzoic acid (1.9g) were dissolved in 10m L DMF and transferred to a Teflon liner after sonication for 3 minutes, placed at 120 ℃ for 24 hours of standing reaction the resulting product was obtained by centrifugation and washed three times with fresh DMF and methanol (8,000rpm, 5 minutes) repeatedly, finally the nanoparticles were collected after drying at 65 ℃ overnight in order to remove residual organic solvent.
Step one, gold nanoparticle preparation, namely, when a chloroauric acid solution (50m L, 1mM) is heated to boiling, a trisodium citrate solution (5m L, 38.8mM) is rapidly added under vigorous stirring, the mixed solution is continuously heated and fully stirred for 30 minutes to ensure that the reaction is finished, and after cooling, the dark red solution is stored at 4 ℃ for use.
Step three: modification of single-stranded DNA by gold nanoparticles
6 different DNA strands (NP 1: SH- (CH)2)6-TTTTT；NP2：SH-(CH2)6-TTTTTTTTTTTTTTT； NP3：SH-(CH2)6-AAAAAAAAAAAAAAA；NP4：SH-(CH2)6-CCCCC；NP5：SH-(CH2)6- CCCCCCCCCCCCCCC；NP6：SH-(CH2)6-GAGAGAGAGAGAGAG) was activated by TCEP (10mM) for 1 hour, then incubated with the prepared AuNPs colloidal solution for 16 hours (final DNA concentration of 3 μ M), after which 0.1M NaCl was gradually added to the mixture to "age" and incubation was continued for 24 hours, then the solution was centrifuged at 13000rpm and washed for 30 minutes to remove excess DNA in the supernatant, finally, the red oily precipitate was dispersed in 1M L Tris-HCl buffer solution (20mM Tris-HCl buffer, containing 100mM naci, pH 7-7.8).
Step four: and (3) detecting a semen sample: ssDNA-AuNPs (NP1-NP6) and Zr-MOF were diluted with Tris-HCl (pH 7-7.8) to the final appropriate concentration. In a 96-well microplate, a certain amount of different ssDNA-AuNPs were transferred to each well, respectively, and the absorption intensity of the solution was recorded at 520nm by means of a Safire microplate reader (A)0) Semen samples of different cases with certain concentrations are added and incubated for 30-120 minutes at 25-40 ℃. (4) Adding a certain amount of Zr-MOF into the mixture solution, and reacting for 2-12 hours in an environment of 25-40 ℃ to combine and precipitate the residual ssDNA-AuNP or protein. The supernatant without precipitation was carefully transferred to a new 96-well microplate and the absorbance intensity (A), the relative absorbance intensity change (A) was measured0-A) is used as the absorbance response change value. The raw data matrix was subjected to linear discriminant analysis using SPSS v25.0 software. As shown in figure 3, the method can easily and effectively make finger print patterns for each type of semen sample according to different semen components, and is expected to be used in medical diagnosis of male infertility.
Claims (5)
1. Preparation and application of colorimetric array sensor of interaction of zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles are characterized by comprising the following steps: zirconium-based metal organic framework, single-stranded DNA modified gold nanoparticles. The zirconium-based metal organic framework comprises one of substances with a molecular formula conforming to UiO-66(Zr) -X, wherein X can be one of-NH 2, -NO2, - (OH)2, -Br and-COOH; (ii) a The size of the zirconium-based metal organic framework is 70-500 nm; the size of the gold nano-particles is 10-200 nm.
2. A preparation method of zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles is characterized by comprising the following steps:
(1) the preparation method of the zirconium-based metal organic framework comprises the steps of dissolving a zirconium metal precursor and an organic ligand in 10m L DMF, transferring the zirconium metal precursor and the organic ligand into a Teflon lining, carrying out ultrasonic treatment, and then carrying out high-temperature standing reaction, wherein the zirconium metal precursor comprises at least one of zirconium tetraoxide, zirconium dichloride, zirconium n-butoxide and zirconium n-propoxide, the concentration of the zirconium metal precursor is 5-25mg/m L, the zirconium metal precursor comprises at least one of zirconium tetraoxide, zirconium dichloride, zirconium n-butoxide and zirconium n-propoxide, the concentration of the zirconium metal precursor is 5-20mg/m L, the reaction temperature is 100-200 ℃, and the reaction time is 10-24 hours.
(2) The preparation method of the gold nanoparticles comprises the steps of mixing 50m L chloroauric acid and 5m L trisodium citrate, heating, reacting and synthesizing the gold nanoparticles, wherein the concentration of the chloroauric acid is 1-5mM, the concentration of the trisodium citrate is 20-50mM, and the reaction time is 10-60 minutes.
(3) The gold nanoparticles are subjected to single-stranded DNA modification: thiolated DNA strands are activated by reaction with tris (2-carboxyethyl) phosphine and incubated with the prepared colloidal solution of AuNPs. Thereafter, 0.1M NaCl was gradually added to the mixture to "age" and incubation was continued. The single-stranded DNA comprises a random combination of thiol-modified Poly A, Poly T, Poly C and A, T, C, G; the length of the single-stranded DNA is 5-50 bp; the concentration of the tri (2-carboxyethyl) phosphine is 1-50 mM; the incubation time is 5-24 hours, and the incubation temperature is 25-40 ℃.
(4) And (4) detecting the protein sample or the semen.
3. A method for using a zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles is characterized by comprising the following steps:
(1) the zirconium-based metal organic framework and single-stranded DNA modified gold nanoparticles were diluted with Tris-HCl (PH 7-7.8) to the final appropriate concentration.
(2) In a 96-well microplate, a certain amount of different single-stranded DNA-modified gold nanoparticles were separately transferred to each well, and the absorption intensity of the solution was recorded at 520nm by a Safire microplate reader (A)0)
(3) Adding different analytes with certain concentration, multiple proteins or different semen samples, and incubating for 30-120 min at 25-40 deg.C.
(4) Adding a certain amount of zirconium-based metal organic framework into the mixture solution, and reacting for 2-12 hours at the temperature of 25-40 ℃ to combine and precipitate the residual single-stranded DNA modified gold nanoparticles or proteins.
(5) The supernatant without precipitation was carefully transferred to a new 96-well microplate and the absorbance intensity (A), the relative absorbance intensity change (A) was measured0-A) is used as the absorbance response change value.
(6) And (4) carrying out linear discriminant analysis on the original data matrix by using corresponding software.
4. The use of claim 3, wherein the colorimetric array sensor of interaction between the zirconium-based metal organic framework and the single-stranded DNA-modified gold nanoparticles can simultaneously perform low-concentration assay detection on a plurality of proteins.
5. The use according to claim 3, wherein the colorimetric array sensor based on interaction of the zirconium-based metal organic framework and the single-stranded DNA modified gold nanoparticles can simultaneously analyze and detect semen from different types of male infertility.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910046219.8A CN111443049B (en) | 2019-01-17 | 2019-01-17 | Preparation method and application of zirconium-based metal organic framework colorimetric array sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910046219.8A CN111443049B (en) | 2019-01-17 | 2019-01-17 | Preparation method and application of zirconium-based metal organic framework colorimetric array sensor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111443049A true CN111443049A (en) | 2020-07-24 |
| CN111443049B CN111443049B (en) | 2022-01-11 |
Family
ID=71626691
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910046219.8A Active CN111443049B (en) | 2019-01-17 | 2019-01-17 | Preparation method and application of zirconium-based metal organic framework colorimetric array sensor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111443049B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114563455A (en) * | 2022-03-04 | 2022-05-31 | 章毅 | DNA-gated metal-organic frameworks and methods of making and using the same |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006078660A2 (en) * | 2005-01-21 | 2006-07-27 | The Board Of Trustees Of The University Of Illinois | Nucleic acid enzyme ligation sensor |
| WO2008017507A2 (en) * | 2006-08-11 | 2008-02-14 | Roche Diagnostics Gmbh | Nanoparticle nucleic acid bonding compound conjugates forming i-motifs |
| CN106483176A (en) * | 2016-09-30 | 2017-03-08 | 重庆医科大学 | A kind of electrochemical sensor preparation method for FGFR3 1138G ＞ A genetic polymorphism detection |
| CN107543852A (en) * | 2017-08-29 | 2018-01-05 | 济南大学 | A kind of Electrochemiluminescsensor sensor based on functional metal organic framework materials |
| CN108387542A (en) * | 2018-05-24 | 2018-08-10 | 清华大学 | Fluorine ion colorimetric sensor and its application in a kind of water based on smart mobile phone |
| CN108760730A (en) * | 2018-05-14 | 2018-11-06 | 济南大学 | A kind of method of paper substrate double mode detection magnesium ion |
-
2019
- 2019-01-17 CN CN201910046219.8A patent/CN111443049B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006078660A2 (en) * | 2005-01-21 | 2006-07-27 | The Board Of Trustees Of The University Of Illinois | Nucleic acid enzyme ligation sensor |
| WO2008017507A2 (en) * | 2006-08-11 | 2008-02-14 | Roche Diagnostics Gmbh | Nanoparticle nucleic acid bonding compound conjugates forming i-motifs |
| CN106483176A (en) * | 2016-09-30 | 2017-03-08 | 重庆医科大学 | A kind of electrochemical sensor preparation method for FGFR3 1138G ＞ A genetic polymorphism detection |
| CN107543852A (en) * | 2017-08-29 | 2018-01-05 | 济南大学 | A kind of Electrochemiluminescsensor sensor based on functional metal organic framework materials |
| CN108760730A (en) * | 2018-05-14 | 2018-11-06 | 济南大学 | A kind of method of paper substrate double mode detection magnesium ion |
| CN108387542A (en) * | 2018-05-24 | 2018-08-10 | 清华大学 | Fluorine ion colorimetric sensor and its application in a kind of water based on smart mobile phone |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114563455A (en) * | 2022-03-04 | 2022-05-31 | 章毅 | DNA-gated metal-organic frameworks and methods of making and using the same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111443049B (en) | 2022-01-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2021200925B2 (en) | Assays for single molecule detection and use thereof | |
| Zheng et al. | Aptamer‐Functionalized Barcode Particles for the Capture and Detection of Multiple Types of Circulating Tumor Cells | |
| Ji et al. | Point‐of‐care identification of bacteria using protein‐encapsulated gold nanoclusters | |
| EP2577300B1 (en) | Scanning multifunctional particles | |
| CN102272597B (en) | Analyte detection assay | |
| Jin et al. | Glass‐bead‐based parallel detection of DNA using composite Raman labels | |
| CN109477095A (en) | Array and its application for Single Molecule Detection | |
| CN109072297A (en) | The method for nucleic acid sequencing and system of detection are formed using the polynucleotide ternary complex of enhancing | |
| CN104792753B (en) | Suppress to click on the biological method for sensing that chemical reaction detects smaller ligand target protein based on combining | |
| Du et al. | Two 3d-4f metal-organic frameworks as fluorescent sensor array for the discrimination of phosphates based on different response patterns | |
| Fang et al. | Review of FRET biosensing and its application in biomolecular detection | |
| Fang et al. | Tuning surface states to achieve the modulated fluorescence of carbon dots for probing the activity of alkaline phosphatase and immunoassay of α-fetoprotein | |
| CN108342459A (en) | A kind of quantitative PCR detecting method based on gold nano grain | |
| Ali et al. | Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats | |
| CN102453761A (en) | Kit for detecting hereditary hearing loss by co-labeling magnetic beads and luminophors | |
| Liu et al. | Recognition engineering-mediated multichannel sensor array for gut microbiota sensing | |
| Zhang et al. | Resettable and enzyme-free molecular logic devices for the intelligent amplification detection of multiple miRNAs via catalyzed hairpin assembly | |
| Zhu et al. | L-tyrosine methyl ester-stabilized carbon dots as fluorescent probes for the assays of biothiols | |
| GB2519467A (en) | Pathogenic microorganism nucleic acid non-amplification detection and classification method | |
| Zhang et al. | Screening prostate cancer cell‐derived exosomal microRNA expression with photothermal‐driven digital PCR | |
| CN111443049B (en) | Preparation method and application of zirconium-based metal organic framework colorimetric array sensor | |
| CN108753925A (en) | A kind of colorimetric detection method and kit of single nucleotide polymorphism | |
| Qiu et al. | Cost-efficient and ultrasensitive hydrogel-based visual point-of-care sensor integrated with surface patterning and strongly emissive carbon dots directly from Prunus mume Carbonisatus | |
| Lin et al. | Gold nanoparticles stabilized with four kinds of amino acid-derived carbon dots for colorimetric and visual discrimination of proteins and microorganisms | |
| CN114410790A (en) | Biosensing detection system for detecting ctDNA and detection method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |