CN110082530A - Aqueogel based on quantum dot and gold nanorods and preparation method thereof, application - Google Patents
Aqueogel based on quantum dot and gold nanorods and preparation method thereof, application Download PDFInfo
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- CN110082530A CN110082530A CN201910281368.2A CN201910281368A CN110082530A CN 110082530 A CN110082530 A CN 110082530A CN 201910281368 A CN201910281368 A CN 201910281368A CN 110082530 A CN110082530 A CN 110082530A
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- quantum dot
- gold nanorods
- carcinomebryonic antigen
- aqueogel
- aptamers
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- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/588—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with semiconductor nanocrystal label, e.g. quantum dots
-
- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/70—Mechanisms involved in disease identification
- G01N2800/7023—(Hyper)proliferation
- G01N2800/7028—Cancer
Abstract
The present invention provides a kind of preparation methods of aqueogel based on quantum dot and gold nanorods, first by quantum dot in conjunction with carcinomebryonic antigen aptamers；Secondly by gold nanorods in conjunction with carcinomebryonic antigen aptamers；The gold nanorods of quantum dot, the modification of carcinomebryonic antigen aptamers that carcinomebryonic antigen aptamers are modified are mixed with hydrogel solution again, finally obtain the aqueogel based on quantum dot and gold nanorods.Preparing aqueogel can detecte carcinomebryonic antigen, and when the carcinomebryonic antigen for being 1 milligram every milliliter with 1-6 microlitres of concentration is incubated for altogether, optical quenching efficiency is 12-24%.
Description
Technical field
The present invention relates to biomedicine technical field, especially a kind of aqueogel based on quantum dot and gold nanorods
And preparation method thereof, application.
Background technique
Operation at present, chemotherapy and radiation are the main means for the treatment of of cancer.Early-stage cancer can be cured by operation excision,
However according to statistics, about ninety percent cancer patient is finally the death because of tumor recurrence or cancer metastasis.This is mainly due to
Before tumor resection, Partial tumors cell is had existed in blood or lymphatic vessel.Although in these years surgical technic obtains
Constantly improve, the postoperative inevitable cancer cell that there are remnants, therefore postoperative recurrence often occurs.However postoperative recurrence early stage in situ is not
As it can be seen that existing means such as B ultrasound, CT etc. can not be detected.At present clinically for monitor product that tumor post-operation recurs in situ compared with
It is few and expensive, very big financial burden is brought to patient.Therefore designing simple and effective detection system is effective prevention of postoperative
The key of recurrence.
Tumor markers are substances existing for a kind of reflection tumour, there is the property that tumour can be prompted with quantitative change, such as
Carcinomebryonic antigen.It can be used as the marker detection had postoperative recurrent tumor of tumor recurrence.
Hydrogel is a kind of macromolecule network system, and property is soft, is able to maintain certain shape.It is widely used in a variety of necks
Domain.Hydrogel has great application prospect in terms of detecting tumor markers as carrier.
Summary of the invention
Technical problem to be solved by the present invention lies in provide a kind of aqueogel based on quantum dot and gold nanorods
And preparation method thereof, application.
In order to solve the above technical problems, first technical solution of the invention is: one kind being based on quantum dot and gold nanorods
Aqueogel preparation method, the specific steps are as follows:
(1) by quantum dot in conjunction with carcinomebryonic antigen aptamers；
(2) by gold nanorods in conjunction with carcinomebryonic antigen aptamers；
(3) gold nanorods and hydrogel of the quantum dot, the modification of carcinomebryonic antigen aptamers modified carcinomebryonic antigen aptamers
Solution mixing, obtains the aqueogel based on quantum dot and gold nanorods.
Preferably, the quantum dot of carboxyl modified is configured to concentration with distilled water in the step (1) is 1-1.5 milligrams every
The solution of milliliter, is added the adaptation liquid solution that 10-20 microlitres of concentration is 50 every liter of micromoles, and magnetic agitation 4-12 hours,
10000-12000 revs/min of centrifugation collection in 5-10 minutes precipitating, precipitating distilled water are resuspended quantitative to 0.5~1 milligram of every milli
It rises, obtains the quantum dot of carcinomebryonic antigen aptamers modification.
Preferably, gold nanorods are configured to concentration with distilled water in the step (2) is 0.5~1 milligram every milliliter
Solution is added the adaptation liquid solution that 10-20 microlitres of concentration is 50 every liter of micromoles, stirs 12-24 hours, 10000-12000 turns/
The minute centrifugation precipitating of collection in 15-30 minutes, precipitating are resuspended quantitatively with distilled water to 0.5~1 milligram every milliliter, obtain carcinomebryonic antigen
The gold nanorods of aptamers modification.
Preferably, quantum dot and carcinomebryonic antigen the aptamers modification modified carcinomebryonic antigen aptamers in the step (3)
Gold nanorods be added in hydrogel solution be uniformly mixed, the hydrogel system based on quantum dot and gold nanorods can be obtained
Agent.
Preferably, the quantum dot and carcinomebryonic antigen aptamers modified carcinomebryonic antigen aptamers in the step (3) are modified
Gold nanorods according to volume ratio 20-50:1 ratio, be added to mass fraction be 20% hydrogel solution in be uniformly mixed.
Second technical solution of the invention is the aqueogel that preparation method obtains, internal load quantum dot and Jenner
Rice stick, when being incubated for altogether with the carcinomebryonic antigen of different volumes, optical quenching efficiency is 12-24%.Third technical solution of the invention
It is the application of aqueogel, the detection preparation of preparation detection tumor markers carcinomebryonic antigen.
The aqueogel that above-mentioned preparation method obtains is for step when detecting carcinomebryonic antigen are as follows:
(1) 1 milliliter of aqueogel is pipetted, is added in glass dish；
(2) the carcinomebryonic antigen standard items that 1-6 microlitres of concentration is 1 milligram every milliliter are added；
(3) it is incubated at room temperature 5 minutes, then excites the nanometer formulation with 450 nanometer lasers, record fluorescence with Fluorescence Spectrometer
Strength Changes.
The aqueogel of above method preparation, can detecte the presence of carcinomebryonic antigen.
The nanometer formulation of above method preparation is used to prepare the detection preparation of detection tumor markers carcinomebryonic antigen.
The nanometer formulation of above method preparation, monitoring probe action principle therein are as follows: in the presence of carcinomebryonic antigen, quantum
The fluorescence intensity of point reduces.
When the aqueogel of above method preparation and the carcinomebryonic antigen of different volumes are incubated for altogether, optical quenching efficiency is 12-
24%.
The beneficial effects of the present invention are:
The preparation method of the above-mentioned aqueogel based on quantum dot and gold nanorods, the aqueogel prepared can be with
Carcinomebryonic antigen is detected, when being incubated for altogether with the carcinomebryonic antigen of different volumes, optical quenching efficiency is 12-24%.
Detailed description of the invention
Fig. 1: the UV absorption figure of the quantum dot of carcinomebryonic antigen aptamers modification；
Fig. 2: the UV absorption figure of the gold nanorods of carcinomebryonic antigen aptamers modification；
Fig. 3: testing result of the aqueogel to carcinomebryonic antigen.
Specific embodiment
Technical solution of the present invention is further described combined with specific embodiments below.
Embodiment 1
(1) preparation of the quantum dot of carcinomebryonic antigen aptamers modification: the quantum dot of carboxyl modified is configured to distilled water
It is suitable that the amido modified carcinomebryonic antigen that 20 microlitres of concentration are 50 every liter of micromoles is added in the solution that concentration is 1.5 milligrams every milliliter
Ligand solution, magnetic agitation 12 hours, 12000 revs/min of centrifugation collection in 10 minutes precipitatings, precipitating was resuspended quantitatively extremely with distilled water
1 milligram every milliliter, obtain the quantum dot of carcinomebryonic antigen aptamers modification.
(2) preparation of the gold nanorods of carcinomebryonic antigen aptamers modification: it is 1 that gold nanorods, which are configured to concentration, with distilled water
The adaptation liquid solution that 20 microlitres of concentration are 50 every liter of micromoles is added in every milliliter of solution of milligram, stirs 24 hours, 12000 turns/
The minute centrifugation precipitating of collection in 30 minutes, precipitating are resuspended quantitatively to 1 milligram every milliliter with distilled water, obtain carcinomebryonic antigen aptamers and repair
The gold nanorods of decorations.
(3) preparation of the aqueogel based on quantum dot and gold nanorods: 1 milliliter of carcinomebryonic antigen aptamers is modified
Quantum dot and the gold nanorods of 50 microlitres of carcinomebryonic antigen aptamers modification are added in the hydrogel solution that mass fraction is 20%
It is uniformly mixed.
Embodiment 2
(1) preparation of the quantum dot of carcinomebryonic antigen aptamers modification: the quantum dot of carboxyl modified is configured to distilled water
It is suitable that the amido modified carcinomebryonic antigen that 15 microlitres of concentration are 50 every liter of micromoles is added in the solution that concentration is 1.2 milligrams every milliliter
Ligand solution, magnetic agitation 8 hours, 11000 revs/min of centrifugation collection in 8 minutes precipitatings, precipitating was resuspended quantitatively extremely with distilled water
0.8 milligram every milliliter, obtain the quantum dot of carcinomebryonic antigen aptamers modification.Pipette the amount of 1 milliliter of carcinomebryonic antigen aptamers modification
It is sub-, be added in glass dish, with Fluorescence Spectrometer excitation wavelength be its fluorescence intensity of 450 nanometer detections, as a result such as Fig. 1 institute
Show.
(2) preparation of the gold nanorods of carcinomebryonic antigen aptamers modification: it is 1 that gold nanorods, which are configured to concentration, with distilled water
The adaptation liquid solution that 15 microlitres of concentration are 50 every liter of micromoles is added in every milliliter of solution of milligram, stirs 18 hours, 11000 turns/
The minute centrifugation precipitating of collection in 20 minutes, precipitating are resuspended quantitatively with distilled water to 0.8 milligram every milliliter, obtain carcinomebryonic antigen aptamers
The gold nanorods of modification.The gold nanorods for pipetting the modification of 1 milliliter of carcinomebryonic antigen aptamers, are added in glass dish, use ultraviolet light
Spectrometer detects its UV absorption, as a result as shown in Figure 2.
(3) preparation of the aqueogel based on quantum dot and gold nanorods: 1 milliliter of carcinomebryonic antigen aptamers is modified
Quantum dot and the gold nanorods of 35 microlitres of carcinomebryonic antigen aptamers modification are added in the hydrogel solution that mass fraction is 20%
It is uniformly mixed.
Embodiment 3
(1) preparation of the quantum dot of carcinomebryonic antigen aptamers modification: the quantum dot of carboxyl modified is configured to distilled water
The amido modified carcinomebryonic antigen adaptation that 10 microlitres of concentration are 50 every liter of micromoles is added in the solution that concentration is 1 milligram every milliliter
Liquid solution, magnetic agitation 4 hours, 10000 revs/min of centrifugation collection in 5 minutes precipitatings, precipitating distilled water was resuspended quantitative to 0.5
Every milliliter of milligram obtains the quantum dot of carcinomebryonic antigen aptamers modification.
(2) preparation of the gold nanorods of carcinomebryonic antigen aptamers modification: gold nanorods, which are configured to concentration, with distilled water is
0.5 milligram every milliliter of solution, be added 10 microlitres of concentration be 50 every liter of micromoles adaptation liquid solution, stir 12 hours, 10000
Rev/min centrifugation precipitating of collections in 15 minutes, precipitating distilled water are resuspended quantitative to 0.5 milligram every milliliter, obtain carcinomebryonic antigen and fit
Ligand modified gold nanorods.
(3) preparation of the aqueogel based on quantum dot and gold nanorods: 1 milliliter of carcinomebryonic antigen aptamers is modified
Quantum dot and the gold nanorods of 20 microlitres of carcinomebryonic antigen aptamers modification are added in the hydrogel solution that mass fraction is 20%
It is uniformly mixed.
Embodiment 4
The hydrogel nanometer formulation for pipetting the preparation of 1 milliliter of embodiment 2, is added in glass dish.1-6 microlitres of concentration, which is added, is
1 milligram every milliliter of carcinomebryonic antigen standard items.Incubation at room temperature 5 minutes, then excites the nanometer formulation with 450 nanometer lasers, uses
Fluorescence Spectrometer records fluorescence intensity change.As a result as shown in Figure 3.
Claims (7)
1. the preparation method of the aqueogel based on quantum dot and gold nanorods, it is characterised in that: specific step is as follows:
(1) by quantum dot in conjunction with carcinomebryonic antigen aptamers；
(2) by gold nanorods in conjunction with carcinomebryonic antigen aptamers；
(3) gold nanorods and hydrogel solution of the quantum dot, the modification of carcinomebryonic antigen aptamers modified carcinomebryonic antigen aptamers
Mixing, obtains the aqueogel based on quantum dot and gold nanorods.
2. the preparation method of the aqueogel according to claim 1 based on quantum dot and gold nanorods, feature exist
In: the quantum dot of carboxyl modified is configured to the solution that concentration is 1-1.5 milligrams every milliliter with distilled water in the step (1),
It is added the adaptation liquid solution that 10-20 microlitres of concentration is 50 every liter of micromoles, magnetic agitation 4-12 hours, 10000-12000 revs/min
Zhongli's heart collection in 5-10 minutes precipitating, precipitating are resuspended quantitatively with distilled water to 0.5~1 milligram every milliliter, and it is suitable to obtain carcinomebryonic antigen
Ligand modified quantum dot.
3. the preparation method of the aqueogel according to claim 1 based on quantum dot and gold nanorods, feature exist
In: gold nanorods are configured to the solution that concentration is 0.5~1 milligram every milliliter with distilled water in the step (2), 10- is added
20 microlitres of concentration are the adaptation liquid solution of 50 every liter of micromoles, are stirred 12-24 hours, 10000-12000 revs/min of centrifugation 15-
The precipitating of collection in 30 minutes, precipitating are resuspended quantitatively with distilled water to 0.5~1 milligram every milliliter, obtain the modification of carcinomebryonic antigen aptamers
Gold nanorods.
4. the preparation method of the aqueogel according to claim 1 based on quantum dot and gold nanorods, feature exist
In: the gold nanorods in the step (3) by quantum dot and carcinomebryonic antigen the aptamers modification that carcinomebryonic antigen aptamers are modified add
Enter into hydrogel solution and be uniformly mixed, the aqueogel based on quantum dot and gold nanorods can be obtained.
5. the preparation method of the aqueogel according to claim 1 based on quantum dot and gold nanorods, feature exist
In: the gold nanorods that the quantum dot that carcinomebryonic antigen aptamers are modified is modified with carcinomebryonic antigen aptamers are pressed in the step (3)
According to the ratio of volume ratio 20-50:1, it is added in the hydrogel solution that mass fraction is 20% and is uniformly mixed.
6. the aqueogel that preparation method according to any one of claim 1 to 5 obtains, it is characterised in that: internal
Quantum dot and gold nanorods are loaded, when being incubated for altogether with the carcinomebryonic antigen of different volumes, optical quenching efficiency is 12-24%.
7. the application of aqueogel according to claim 6, for detecting tumor markers carcinomebryonic antigen.
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