CN105036073B - Preparation method of gold nucleus-quantum dot satellite structure assembly - Google Patents
Preparation method of gold nucleus-quantum dot satellite structure assembly Download PDFInfo
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Abstract
The invention discloses a preparation method of a gold nucleus-quantum dot satellite structure assembly, belonging to the technical field of analytical chemistry. A high-yield assembly method of a gold nucleus-quantum dot satellite structure comprises the following steps of synthesis of gold nano particles with the particle size of 35nm, preparation of quantum dots with the particle size of 7nm, DNA-1 modification on the gold nano particles, complementary sequence DNA-2 modification on the quantum dots, assembling of the gold nucleus-quantum dot satellite structure, and purification of the gold nucleus-quantum dot satellite structure. By adopting the preparation method, the problems that quantum dot assembling is difficult and the yield is low are solved, and a way is paved for developing a fluorescence sensor based on quantum dot assembling.
Description
Technical field
The present invention relates to a kind of high yield assemble method of gold core-quantum dot satellite structure, belong to technique of analytical chemistry neck
Domain.
Background technology
Quantum dot (quantum dots, qds) is a kind of nano-particle elementary composition by ii-vi race or iii-v race.Amount
The particle diameter of son point is typically in the range of between 1～10nm, and because electronics and hole are by quantum confinement, continuous band structure becomes and has
The discrete energy level structure of molecular characterization, can launch fluorescence after being excited.Compare with conventional fluorescent dyestuff, qds have width excite, narrow
Transmitting and high fluorescence quantum yield.Qds has gradually replaced dyestuff as a kind of new biological labled material, is widely used in
Cell imaging, immunofluorescence technique, living imaging, nucleic acid recognizing aspect, are realizing high sensitivity, high-throughout biology point simultaneously
Analysis and context of detection show substantial promise.
Golden nanometer particle has good biocompatibility, and specific surface area is big.Using its quenching effect to quantum dot, can
To build the sensor with fluorescence as signal.But the assembling of current quantum dot is difficult a lot, leads to assembling yield very low, hinder
Its application in terms of sensing.
Modified by dna hybridization and the golden nanometer particle of big particle diameter and quantum dot are assembled into nuclear satellite structure, have well
Detection Application in Sensing prospect.
Content of the invention
It is an object of the invention to provide a kind of high yield assemble method of gold core-quantum dot satellite structure.
Technical scheme, a kind of preparation method of gold core-quantum dot satellite structure assembly, step is:
The synthesis of the golden nanometer particle of (1) 35 nm particle diameter: the golden nanometer particle of 35 nm particle diameters adopts seed mediated growth method
Synthesis, the gold seeds of 13nm adopts trisodium citrate to reduce the method synthesis of gold chloride；
The conical flask of reaction uses the new chloroazotic acid prepared to soak 24h in advance, and with ultrapure water to cleaning, 37 DEG C of drying are treated
With.The gold chloride of 2ml 10mm, 38ml ultra-pure water add in clean conical flask, are heated to seething with excitement, add 2ml 38.8mm's
Trisodium citrate, boils the gold seeds that 20min obtains 13nm particle diameter.
The synthesis of the golden nanometer particle of 35nm particle diameter: add the gold chloride of 2ml 10mm, 0.1ml 10mm in conical flask
Silver nitrate, 42.5ml ultra-pure water, the gold seeds of 2ml 13nm particle diameter, add the ascorbic acid of 7.5ml 5.3mm under stirring
Solution is pumped into the speed of 30ml/h, is sufficiently stirred for 20s, set aside for use, obtains the golden nanometer particle of diameter 35nm particle diameter.
(2) CdSe quantum dots of the carboxyl water-soluble ZnS parcel of the preparation of the quantum dot of 7nm particle diameter: 7nm
(gdse zns) buys in Wuhan Jia Yuan quantum dot company limited.
(3) on golden nanometer particle dna-1 modification: the golden nanometer particle of 35nm particle diameter adopt 13nm particle diameter gold seeds life
Long synthesis.
Ten times of the golden nanometer particle centrifugal concentrating of 35 nm particle diameters uses the phosphate buffer of 10mm ph7.2 resuspended afterwards, plus
The dna-1 entering sulfhydrylation makes final concentration of 5 μm.Dna-1 is 300:1 with the mol ratio of golden nanometer particle.After incubated at room 2h
Plus nacl to 50mm, afterwards every 1h plus nacl, to final concentration 500mm.It is centrifuged 3 removals after solution age 24h not tying
The dna-1 closing, the phosphate buffer of 10mm ph7.2 is resuspended stand-by；
Dna-1:5 '-caatagccct tggatttttt ttttt-sh-3 '；
(4) on quantum dot complementary seriess dna-2 modification: 7nm carboxyl water-soluble ZnS parcel cadmium selenide quantum
Point (gdse zns) is bought in Wuhan Jia Yuan quantum dot company limited.Quantum dot is diluted to the borate buffer solution of 10mm ph8
100nm, adds amidized complementary seriess dna-2 to make final concentration of 0.5 μm, and addition edc(1- (3- dimethylamino-propyl)-
3- ethyl-carbodiimide hydrochloride) make final concentration reach 0.5 mm, plus nhs(n- hydroxysuccinimide) make final concentration reach 0.5mm,
It is put in reaction 2h on agitator.Ultrafiltration removes unconjugated dna-2, resuspended stand-by with the phosphate buffer of 10mm ph7.2.
Dna-2:5 '-atccaagggc tattgttttt ttttt-nh2-3’；
(5) assembling of golden core-quantum dot satellite structure: the golden nanometer particle that 50 μ l aptamers dna-1 are modified is mutual with 200 μ l
The quantum dot mixing that complementary series dna-2 modifies, 60 DEG C of incubation 2h, obtain golden core-quantum dot satellite structure assembly.
(6) purification of golden core-quantum dot satellite structure assembly: by assembly gradient centrifugation purification, 1000rpm is centrifuged
3 min, abandon precipitation.Supernatant 3500rpm is centrifuged 10 min, collects precipitation, be resuspended in the phosphate buffer of 10mm ph7.2
In.
Beneficial effects of the present invention: the invention provides a kind of assembling side of the golden core-quantum dot satellite structure of high yield
Method, solves the problems, such as quantum dot assembling difficulty, low yield, has paved road for exploitation based on the fluorescent optical sensor of quantum dot assembling
Road.
Brief description
Fig. 1 is the transmission electron microscope photo of the golden core-quantum dot satellite structure assembly being formed by nucleic acid hybridization.
Specific embodiment
The high yield assembling of embodiment 1 gold medal core-quantum dot satellite structure
The synthesis of the golden nanometer particle of (1) 35 nm particle diameter
The golden nanometer particle of 35 nm particle diameters adopts seed mediated growth method to synthesize.
The gold seeds of 13 nm particle diameters adopts trisodium citrate to reduce the method synthesis of gold chloride: the conical flask of reaction carries
The front new chloroazotic acid prepared soaks 24h, and extremely clean with ultrapure water, 37 DEG C of drying are stand-by.The gold chloride of 2ml 10mm, 38ml
Ultra-pure water adds in clean conical flask, is heated to seething with excitement, adds the trisodium citrate of 2ml 38.8mm, boil 20min and obtain
The gold seeds of 13nm particle diameter.
The synthesis of the golden nanometer particle of 35nm particle diameter: add the gold chloride of 2ml 10mm, 0.1ml 10mm in conical flask
Silver nitrate, 42.5ml ultra-pure water, the gold seeds of 2ml 13 nm particle diameter, add the ascorbic acid of 7.5ml 5.3mm under stirring
Solution is pumped into the speed of 30ml/h, is sufficiently stirred for 20s, set aside for use, obtains the golden nanometer particle of diameter 35nm particle diameter.
The preparation of the quantum dot of (2) 7 nm particle diameters
The CdSe quantum dots (gdse@zns) of the carboxyl water-soluble ZnS parcel of 7 nm are bought in Wuhan Ka source quantum
Point company limited.
(3) on golden nanometer particle dna-1 modification
The golden nanometer particle of 35 nm particle diameters adopts the gold seeds growth synthesis of 13 nm particle diameters.The Jenner of 35 nm particle diameters
Ten times of rice corpuscles centrifugal concentrating uses the phosphate buffer of 10mm ph7.2 resuspended afterwards, adds the dna-1 of sulfhydrylation to make dense eventually
Spend for 5 μm.Dna-1 is 300:1 with the mol ratio of golden nanometer particle.Nacl to 50mm is added, afterwards every 1h after incubated at room 2h
Plus a nacl, to final concentration 500mm.After solution age 24h, centrifugation removes unconjugated dna-1, the phosphorus of 10mm ph7.2 for 3 times
Phthalate buffer is resuspended stand-by
Dna-1:5 '-caatagccct tggatttttt ttttt-sh-3 '；
(4) on quantum dot complementary seriess dna-2 modification
The CdSe quantum dots (gdse@zns) of the carboxyl water-soluble ZnS parcel of 7 nm are bought in Wuhan Ka source quantum
Point company limited.The quantum dot borate buffer solution of 10mm ph8 is diluted to 100 nm, adds amidized complementary seriess
Dna-2 makes final concentration of 0.5 μm, adds edc(1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride) make end dense
Degree reaches 0.5 mm, plus nhs(n- hydroxysuccinimide) make final concentration reach 0.5 mm, it is put in reaction 2h on agitator.Ultrafiltration is gone
Except unconjugated dna-2, resuspended stand-by with the phosphate buffer of 10mm ph7.2.
Dna-2:5 '-atccaagggc tattgttttt ttttt-nh2-3’
(5) assembling of golden core-quantum dot satellite structure
The quantum dot that the golden nanometer particle that 50 μ l aptamers dna-1 are modified is modified with 200 μ l complementary seriess dna-2 mixes,
60 DEG C of incubation 2h, obtain golden core-quantum dot satellite structure assembly.
(6) purification of golden core-quantum dot satellite structure assembly
By assembly gradient centrifugation purification, 1000rpm is centrifuged 3 min, abandons precipitation.Supernatant 3500 rpm is centrifuged 10
Min, collects precipitation, is resuspended in the phosphate buffer of 10mm ph7.2.
Claims (1)
1. a kind of preparation method of gold core-quantum dot satellite structure assembly is it is characterised in that step is:
(1) on golden nanometer particle dna-1 modification: the golden nanometer particle of 35nm particle diameter adopt 13nm particle diameter gold seeds growth close
Become；
Use the phosphate buffer of 10mm ph7.2 resuspended after the golden nanometer particle centrifugal concentrating of 35 nm particle diameters, add sulfhydrylation
Dna-1 make final concentration of 5 μm of the dna-1 of sulfhydrylation；Dna-1 is 300:1 with the mol ratio of golden nanometer particle, incubated at room
Nacl to 50mm is added, afterwards every 1h plus nacl, to the final concentration of 500mm of nacl after 2h；It is centrifuged 3 after solution age 24h
The unconjugated dna-1 of secondary removal, the phosphate buffer of 10mm ph7.2 is resuspended stand-by；
Dna-1:5 '-caatagccct tggatttttt ttttt-sh-3 '；
(2) on quantum dot complementary seriess dna-2 modification: 7nm carboxyl water-soluble ZnS parcel CdSe quantum dots use
The borate buffer solution of 10mm ph8 is diluted to 100nm, adds amidized complementary seriess dna-2 to make dna-2 final concentration of
0.5 μm, 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride is added to be that edc makes edc final concentration reach 0.5mm, plus n-
Hydroxysuccinimide is that nhs makes nhs final concentration reach 0.5mm, is put in reaction 2h on agitator；Ultrafiltration removes unconjugated dna-
2, resuspended stand-by with the phosphate buffer of 10mm ph7.2；
Dna-2:5 '-atccaagggc tattgttttt ttttt-nh2-3’ ；
(3) assembling of golden core-quantum dot satellite structure: golden nanometer particle and 200 μ l complementation sequence that 50 μ l aptamers dna-1 are modified
The quantum dot mixing that row dna-2 modifies, 60 DEG C of incubation 2h, obtain golden core-quantum dot satellite structure assembly；
(4) purification of golden core-quantum dot satellite structure assembly: by assembly gradient centrifugation purification, 1000rpm is centrifuged
3min, abandons precipitation；Supernatant 3500rpm is centrifuged 10 min, collects precipitation, be resuspended in the phosphate buffer of 10mm ph7.2
In.
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| CN105567807B (en) * | 2015-12-16 | 2019-06-25 | 苏州大学 | The detection method of microRNA molecule in a kind of novel living cells |
| CN105834452B (en) * | 2016-05-30 | 2017-07-25 | 江南大学 | A kind of preparation method of chiral adjustable ghost satellite shape structure nano assembly |
| CN111974985B (en) * | 2020-09-16 | 2022-03-01 | 南京大学 | Nano particle cluster assembling method using micro magnetic beads as growth template and DNA frame as guide carrier |
| CN112643043A (en) * | 2020-12-02 | 2021-04-13 | 杭州苏铂科技有限公司 | Preparation method of aptamer modified gold nano-star |
| CN113376130B (en) * | 2021-05-14 | 2024-06-14 | 南京师范大学 | Fluorescent open probe for detecting ampicillin residue, and preparation method and application thereof |
| CN113433111A (en) * | 2021-06-25 | 2021-09-24 | 辽宁大学 | Liposome-based core-shell material for Raman spectrum detection and preparation method and application thereof |
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| CN101993467A (en) * | 2009-08-24 | 2011-03-30 | 香港科技大学 | Method of manipulating the surface density of functional molecules on nanoparticles |
| CN102556959A (en) * | 2011-12-30 | 2012-07-11 | 中国科学院苏州纳米技术与纳米仿生研究所 | Preparation method of metal nanoparticle dimer |
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