CN109705841A - A kind of transferrins is the gold nano cluster and its preparation method and application of template - Google Patents
A kind of transferrins is the gold nano cluster and its preparation method and application of template Download PDFInfo
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Abstract
The present invention provides the gold nano cluster and its preparation method and application that a kind of transferrins is template.The preparation method of the nanocluster: HAuCl is respectively configured4With the aqueous solution of Tf, 20-60:1 mixes them in molar ratio, then it is stirred at room temperature 2-5 minutes, after the pH of mixed solution is adjusted to 12 with NaOH solution, solution is placed in microwave reactor and is sealed, at least 50min is reacted at 70-90 DEG C and obtains the AuNCs@Tf of intense red fluorescent emission.Nanocluster of the present invention can be used to detect copper ion, and be applied to copper ion test paper, and the detection of copper ion is visualized.Nanocluster of the present invention can also be used to detect glutathione, and show that it can target lysosome to identify cancer cell by cell experiment.Nanocluster of the present invention can also be used for the encryption and decryption of finger print information.
Description
Technical field
The present invention relates to fluorescence detection material, specifically a kind of transferrins be template red fluorescence gold nano cluster and
Preparation method and application.
Background technique
Copper ion (Cu2+) it is the mankind and one of the microelement in other mammals, it is sent out in our health
Wave key effect.Cu2+Intake will cause intestinal disorder and liver or kidney damage and several nervus retrogression diseases extremely
Disease, such as stages alzheimer's disease, Parkinson's disease and Wilson's disease.In addition, industrial excess emissions will also result in environmental pollution.Therefore
A kind of method for finding sensitive detection copper ion, the detection polluted for food and environment copper ion have important value.
Glutathione (GSH) is that pass is played in many physiology and pathologic process containing sulfydryl and γ-amido bond tripeptides
Key effect.Each cell of body is substantially all containing glutathione, and glutathione is mainly by glutamic acid, cysteine and sweet ammonia
Acid composition.The GSH (cytoplasm (2-10mM) and blood plasma (2-20 μM)) of normal level, which has, maintains normal immune system, antioxygen
Change and comprehensive detoxication.However, excessive GSH with include include cancer, leucocyte loss, diabetes and human immune deficiency
Disease including virus is related.On the other hand, it was reported that GSH shortage will lead to aging, liver diseases and neurodegenerative disease.
It is interesting that the tumor tissues being previously reported have more reducing environments than normal tissue, and their GSH concentration ratio is just
Often tissue is several times high.Therefore, detection GSH level is most important.
Currently, there are many technologies for measuring GSH, including high performance liquid chromatography, Surface enhanced Raman scattering, fluorescence light
Spectrum, enzyme linked immunosorbent assay (ELISA), electrochemistry voltammetry, UV, visible light method and capillary electrophoresis.However, these technologies are usually high
It is expensive, it is complicated, time-consuming and impracticable for biosystem application.Recently, people increasingly pay close attention to for the glimmering of GSH detection
The exploitation of light probe, with easy to operate, reaction condition is mild, high sensitivity, responds fast advantage.It is disappointed to be,
Cysteine (Cys) in mammalian cell, the similar structures and property of homocysteine (Hcy) and glutathione (GSH)
Several challenges are brought for selectivity and the identification detection of each individual mercaptan.So far, people explore many glimmering
Light probe, especially fluorescent dye.However, most of organic dyestuff have narrow excitation spectrum, and photostability is poor, cell
Toxicity and environmental risk, which has limited their uses.In addition, also developed carbon quantum dot fluorescence probe (MnO2- CDs) it uses
In identification GSH.Unfortunately, the quantum dot reaction condition containing heavy metal is very harsh and has potential cytotoxicity.With this
A little materials are compared, and gold nano cluster has a big Stokes shift, good biocompatibility, and in vitro and in vivo at
Image space face shows huge potentiality.
Based on this, the present invention restored under microwave condition with transferrins and be coated with tetra chlorauric acid obtain hair intense red it is glimmering
The gold nano cluster of light, and using photoluminescent property be applied to detection copper ion, detect GSH, targeting lysosome identification cancer cell with
And the encryption and decryption of finger print information.
Summary of the invention
It is the preparation method of the red fluorescence gold nano cluster of template the purpose of the present invention is to provide a kind of transferrins,
The preparation method is simple, quick；Nanocluster obtained can detect copper ion, detect GSH, and targeting lysosome identifies cancer cell
And the encryption and decryption of finger print information.
A kind of transferrins provided by the invention is the preparation method of the red fluorescence gold nano cluster of template, including as follows
Step:
HAuCl is respectively configured4With the aqueous solution of Tf, 20-60:1 mixes them in molar ratio, is then stirred at room temperature
2-5 minutes, after the pH of mixed solution is adjusted to 12 with NaOH solution, solution is placed in microwave reactor and is sealed, 70-
At least 50min is reacted at 90 DEG C obtains the AuNCs@Tf of intense red fluorescent emission.It is spare that it is put into 4 DEG C of refrigerators later.
The HAuCl4Molar ratio with Tf is preferably 40:1.
Reaction condition is preferably 60min at 80 DEG C in the microwave reactor.
The red fluorescence gold nano cluster of above-mentioned preparation can be applied in detection copper ion and glutathione, can also be in target
It is applied in the encryption and decryption for identifying cancer cell and finger print information to lysosome.
Beneficial effects of the present invention compared with prior art: fluorescence gold nano cluster good biocompatibility of the invention, it is glimmering
Light intensity, reaction condition green non-pollution；Compared with the research of early period, transferrins ratio used in the present invention is low, the reaction time
It is short, and by production copper ion test paper, the detection of copper ion is visualized；Due to non-luminous AuNCs@Tf-Cu2+In GSH
Under the action of AuNCs@Tf release to making fluorescence restore, and content of the GSH in tumour cell is much higher than normal cell.It utilizes
This property, the AuNCs@Tf-Cu that will be quenched2+It is separately added into normal cell and cancer cell, the only fluorescence of cancer cell restores, and
Normal cell does not change, and distinguishes normal cell and cancer cell with this.In addition, mixed culture normal cell and cancer cell, only
Having has fluorescence to also turn out that gold nano cluster of the invention has identity to cancer cell in cancer cell, mention for the early diagnosis of cancer
A kind of simple and easy method is supplied.
Detailed description of the invention
Fig. 1 is influence of the different preparation conditions to AuNCs@Tf；Wherein: influence of the molar ratio to AuNCs@Tf A, is reacted,
B, influence of the reaction time to AuNCs@Tf, the influence of C, reaction temperature to AuNCs@Tf, the shadow of D, reaction pH to AuNCs@Tf
It rings.
Fig. 2 is the characterization of AuNCs@Tf；Wherein: A, the ultraviolet and fluorogram of AuNCs@Tf, B, AuNCs@Tf it is infrared
Analysis of spectra, the transmission electron microscope analysis figure (the granularmetric analysis figure of illustration AuNCs@Tf) of C, AuNCs@Tf, the X of D, AuNCs@Tf
X ray diffraction analysis x spectrogram, E, X-ray photoelectron spectroscopic analysis spectrogram, the X-ray photoelectron spectroscopic analysis spectrogram of F, Au 4f.
Fig. 3 is selectivity and sensitivity of the AuNCs@Tf to copper ion；Wherein A, AuNCs@Tf and each metal ion are mutual
Effect relative fluorescence compares figure, B, various concentration copper ion be added to the fluorescence emission spectrum of AuNCs@Tf, C, copper ion are dense
The linear relationship chart with fluorescence intensity is spent, the copper ion action diagram of AuNCs@Tf test paper and various concentration under D, ultraviolet lamp.
Fig. 4 is AuNCs@Tf-Cu2+To the selectivity and sensitivity of GSH；Wherein A, various concentration GSH be added to
AuNCs@Tf-Cu2+Fluorescence emission spectrum, the linear relationship chart of B, glutathione concentrations and fluorescence intensity, C, AuNCs@Tf-
Cu2+The figure compared with biomolecule and ionic interaction relative fluorescence, D, homocysteine (Hcy, 15 μM) and cysteine
GSH (150 μM) is in AuNCs@Tf-Cu in the presence of (Cys, 15 μM)2+In fluorescence intensity.
Fig. 5 various concentration AuNCs@Tf is on human cervical carcinoma cell (HeLa) active influence.
Fig. 6 confocal laser scanning microscope AuNCs@Tf and lysosome common location figure.
Fig. 7 AuNCs@Tf-Cu2+For GSH sensing and cancer cell identification in lysosome；Wherein A, use fluorescence microscope
AuNCs@Tf-Cu2+With human cervical carcinoma cell (HeLa), human liver cancer cell (HepG2), human colon cancer cell (HCT 116) and small
Mouse fibroblast cell (3T3) acts on different time figure, B, AuNCs@Tf-Cu2+It is strong from the relative fluorescence of different cell incubation times
Degree figure (data come from Fig. 7 A), C, AuNCs@Tf-Cu2+Into the relative rate constant of different cells, the people palace of D, number ratio 1:1
Neck cancer cell (HeLa) and l cell (3T3) are mixed, and AuNCs@Tf-Cu is added2+Fluorescence microscope is used after 1h
The figure of observation.
Fig. 8 AuNCs@Tf and different cytosis figures；A, with fluorescence microscope AuNCs@Tf and human cervical carcinoma cell
(HeLa), human liver cancer cell (HepG2), when human colon cancer cell (HCT 116) is different with l cell (3T3) effect
Between scheme, the relative intensity of fluorescence figure (data come from Fig. 8 A) of B, AuNCs@Tf and different cell incubation times, C, AuNCs@Tf into
Enter the relative rate constant of different cells.
AuNCs@Tf on Fig. 9 filter paper is sequentially added into Cu2+With the photo and fluorescent image of GSH；Wherein A, Chinese character, B, refer to
Line.
Specific embodiment
The following are materials used in embodiment:
(Holo-Transferrin human, molecular weight are 7.7 × 10 to transferrins4) it is Sigma reagent Co., Ltd
Production；
Tetra chlorauric acid (HAuCl4·4H2O, molecular weight 411.9) it is that fuzz chemical reagent factory in Shanghai produces；
Sodium hydroxide (NaOH, molecular weight 40.0) is Beijing Chemical Plant's production；
Glutathione (C10H17N3O6S, molecular weight 307.33) work of making a living biological production；
Various biomolecule, small molecule and ion (cysteine Cys, homocysteine Hcy, phenylalanine PHE, rely
Propylhomoserin Lys, tyrosine Tyr, glucose glucose, ascorbic acid VC, potassium ion K+, sodium ion Na+, calcium ion Ca2+, sulphur from
Sub- S2-, iodide ion I-, oxalate denominationby C2O4 2-, cyanide ion CN-)
Trishydroxymethylaminomethane (C4H11NO3, molecular weight 121.14) and it is that the life of fine chemistry industry research institute is recovered in Tianjin
It produces.
Embodiment 1
Transferrins, which is restored and is coated with, prepares red fluorescence gold nano cluster.
The transferrins (Tf) of 5mg is weighed, lmL bis- is added wherein and steams water, is made into the Transferrin solution of 5mg/mL.Claim
Take 206.0mg tetra chlorauric acid (HAuCl4·4H2O), the HAuCl of 0.01M concentration is prepared in 50mL brown volumetric flask4Solution is simultaneously
It is spare to place 4 DEG C of refrigerator cold-storages.Prepare the NaOH solution 10mL of 1M.Take the 0.01M HAuCl of 1mL4Solution steams water dilution with two
Four times.Solution after taking 1mL to dilute, is put into microwave tube, the Transferrin solution that 1mL is added is stirred at room temperature 5 minutes.With
After the pH of mixed solution is adjusted to 12 by the 1M NaOH solution of 40 μ L, solution is placed in microwave reactor and is sealed.It will reaction
Condition setting is 80 DEG C, 60min, 150W, 250psi, high mixing speed, and pre- mixing time is 2min.4 are put it into after reaction
DEG C refrigerator is spare, and (Fig. 1 is to change HAuCl when preparing material4It is optimal anti-with determination with the molar ratio of Tf, temperature, time, pH
Answer condition).
Embodiment 2
(1) gold nano cluster AuNCs@Tf is ultraviolet and Fluorescent Characterization.
It is observed in Fig. 2A illustration, brown is presented in the material of synthesis under fluorescent light, has strongly red in the UV lamp
Color fluorescence.It takes AuNCs@Tf solution made from 200 μ L 800 μ L bis- are added to steam water to be diluted, measures ultraviolet absorption curve and glimmering
Light emitting curve.UV absorption has apparent absorption at 280nm as the result is shown, and fluorescence spectrum shows that best emission peak exists
640nm。
(2) infrared analysis (FTIR) spectral characterization of gold nano cluster AuNCs@Tf.
In order to determine that transferrins is incorporated in the surface AuNCs@Tf, by obtained AuNCs@Tf liquid by freeze-drying
Device is dried to obtain solid.Take transferrins and potassium bromide respectively is 1:100 mixed grinding tabletting in mass ratio, measures its infrared light
Spectrum；Then take obtained AuNCs@Tf and potassium bromide is 1:100 mixed grinding tabletting in mass ratio, measures its infrared spectroscopy.
In fig. 2b, their spectrum almost cannot be distinguished, and show that transferrins is already fixed on the surface of AuNCs.
The stretching vibration of the C=O and N-H of transferrins are respectively 1654cm-1And 1540cm-1, and the peak displacement of AuNCs@Tf, this
Kind transformation is mainly the variation of the transferrins skeleton structure as caused by the interaction of Tf and Au.AuNCs@Tf is in 881cm-1
And 798cm-1Place shows new peak, this peak may be attributed to the unsaturated carbon-carbon double bond of similar quinone.This may be due to turning
Tyrosine residue (enol structure) in ferritin is by HAuCl4It is oxidized to quinone.
(3) the transmission electron microscope analysis spectrogram and granularmetric analysis spectral characterization of gold nano cluster AuNCs@Tf.
In order to confirm gold nano cluster morphology and size size, the ultrasonic half an hour drop of AuNCs@Tf solution is made on copper mesh
Sample is observed wait evaporate with transmission electron microscope.Furthermore the AuNCs@Tf liquid of ultrasound is put into Malvern ParticleSizer and is surveyed
Measure partial size.
Fig. 2 C is the transmission electron microscope analysis spectrogram of prepared AuNCs@Tf, as can be seen from Figure obtained AuNCs@Tf
It is uniformly dispersed and in spherical, partial size is 4.75 ± 0.5nm.Illustration is the granularmetric analysis spectrogram of prepared AuNCs@Tf, as a result
6.25 ± 0.7nm of hydraulic diameter, it is consistent with the result of lens Electronic Speculum.
(4) X-ray diffraction analysis (XRD) spectral characterization of gold nano cluster AuNCs@Tf.
In order to observe the crystal form state of obtained AuNCs@Tf, by obtained AuNCs@Tf fluid sample by freezing
Drier is dried to obtain solid, and grinds sample preparation measurement.
In figure 2d, the characteristic diffraction peak of AuNCs@Tf is respectively 31.5 °, 45.2 °, 68.7 ° and 75.2 °, is respectively corresponded
In lattice plane (111), (200), (220) and (311).This shows that AuNCs@Tf is in face-centered cubic (fcc) structure.
(5) X-ray photoelectron spectroscopic analysis (XPS) spectral characterization of gold nano cluster AuNCs@Tf.
In order to confirm functional group and the element on the surface AuNCs@Tf, obtained fluid sample is done by freeze-dryer
It is dry to obtain solid, it is characterized on X-ray photoelectron spectroscopic analysis instrument.
As shown in Figure 2 E, there are five types of elements A u, S, O, C and N on AuNCs@Tf.The XPS of Au 4f spectrum as shown in Figure 2 F
Spectrum is confirmed the existence of two different Au 4f7/2It is bimodal, one in 84.4eV, another is respectively belonging to Au (0) in 87.8eV
With Au (I).It is 1.51 for AuNCs@Tf, Au (1)/Au (0) ratio, it means that Au (I) (principal mode of Au-S key)
It is 60.2% of all Au atoms in AuNCs@Tf.It has been recognized that the aggregation of Au (I)-thiolate complex is Jenner
The main reason for rice cluster (AuNCs) shines.
Embodiment 3
The fluorescence of the interaction of various ions and the gold nano cluster AuNCs@Tf of synthesis is studied.
Prepare each solion 2mL (Mg of 0.1M2+,Pb2+,K+,Cd2+,Cr3+,Bi3+,Zn2+,Na+,Al3+,Ca2+,Ag+,
Mn2+, Ba2+,Cu2+,Hg2+,Co2+,Ni2+) in the EP pipe of 2mL, AuNCs@Tf solution made from 1mL is taken, pH is added thereto
=4.5 Tris-HCl solution 9ml is diluted.Parameter (the λ of Fluorescence Spectrometer is setex=410nm, λem=570nm-
740nm), it takes 1mL to be placed in fluorescence cuvette and scans sample, record data；10 μ L Mg are added into fluorescence cup2+Solution, into
After row stirring, timing 2min scans sample, records data, and other ions repeat above-mentioned experiment.Fig. 3 A has recorded experimental result,
As a result Cu is proved2+It can make the fluorescent quenching of AuNCs@Tf.
Embodiment 4
The fluorescence of the interaction of copper ion and the gold nano cluster AuNCs@Tf of synthesis is studied.
Prepare the Cu of various concentration2+Solution.AuNCs@Tf solution made from 1mL is taken, is added pH=4.5's thereto
Tris-HCl solution 9ml is diluted.Parameter (the λ of Fluorescence Spectrometer is setex=410nm, λem=570nm-740nm), it takes
1mL, which is placed in fluorescence cuvette, scans sample, records data；10 μ L Cu are added into fluorescence cup2+Solution, after being stirred, meter
When 2min, scan sample, record data, by the Cu of various concentration2+Solution repeats above-mentioned experiment.Fig. 3 B, 3C the result shows that, one
(0-2300nM), Cu are determined in concentration range2+Concentration and relative intensity of fluorescence at good linear relationship, and detection is limited to
232nM。
Embodiment 5
Cu2+The production of fluorescent test paper.
AuNCs@Tf is impregnated and filter paper and is dried up, then by filter paper be dipped into various concentration copper ion (10 μM, 20 μM, 40 μ
M, 100 μM, 200 μM, 400 μM) in drying, excited in 365nm ultraviolet lamp, as shown in Figure 3D, available various concentration
Cu2+After AuNCs@Tf effect, AuNCs@Tf-Cu2+The variation of fluorescence, by Cu2+Measurement visualization.
Embodiment 6
Glutathione and AuNCs@Tf-Cu2+The fluorescence of the interaction of system is studied.
Prepare the glutathione solution of various concentration.AuNCs@Tf solution made from 1mL is taken, pH=4.5 is added thereto
Tris-HCl solution 9ml be diluted, the Cu of 100 μ L 0.04M is added thereto2+Solution is stirred.Fluorescence light is set
Parameter (the λ of spectrometerex=410nm, λem=570nm-740nm), it takes above-mentioned solution 1mL to be placed in fluorescence cuvette and scans sample,
Record data；10 μ L glutathione solutions are added into fluorescence cup, after being stirred, timing 2min scans sample, records number
According to；The glutathione solution of various concentration is repeated into above-mentioned experiment.Fig. 4 A, 4B the result shows that, (the 0- within the scope of a certain concentration
150 μM), the concentration and relative intensity of fluorescence of glutathione are at good linear relationship, and detection is limited to 2.86 μM.
Embodiment 7
AuNCs@Tf-Cu2+Selectivity of the system to GSH.
Prepare the various biomolecule, small molecule and solion 2mL (cysteine Cys, homocysteine of 0.01M
Hcy, phenylalanine PHE, lysine Lys, tyrosine Tyr, glucose glucose, ascorbic acid VC, potassium ion K+, sodium ion
Na+, calcium ion Ca2+, sulphion S2-, iodide ion I-, oxalate denominationby C2O4 2-, cyanide ion CN-).It takes made from 1mL
AuNCs@Tf solution, the Tris-HCl solution 9ml that pH=4.5 is added thereto are diluted, and 100 μ L 0.04M are added thereto
Cu2+Solution is stirred.Parameter (the λ of Fluorescence Spectrometer is setex=410nm, λem=570nm-740nm), it takes above-mentioned molten
Liquid 1mL, which is placed in fluorescence cuvette, scans sample, records data；10 μ L cysteine solutions are added into fluorescence cup, are stirred
After mixing, timing 2min scans sample, records data；Continue to repeat to titrate, until fluorescence no longer changes, saves data.Other lifes
Object small molecule repeats above-mentioned experiment.Fig. 4 C has recorded experimental result, as a result proves AuNCs@Tf-Cu2+There is choosing to glutathione
Selecting property.
Further, since intracellular GSH content is 10 times of Cys, we have detected AuNCs@Tf-Cu2+In other biological sulphur
To the influence of GSH detection in the presence of alcohol.It takes the above-mentioned solution 1mL being quenched to be placed in fluorescence cuvette and scans sample, record data；
10 μ L 15.3mM glutathione solutions and 10 μ L 1.53mM homocysteine solution are added into fluorescence cup, are stirred
Afterwards, timing 2min scans sample, records data；The above-mentioned solution 1mL being quenched is taken to be placed in fluorescence cuvette, into fluorescence cup
10 μ L 15.3mM glutathione solutions and 10 μ L 1.53mM cysteine solutions are added, after being stirred, timing 2min, scanning
Sample records data.As shown in Figure 4 D, the presence of Hcy (0.1 equivalent) or Cys (0.1 equivalent) will not interfere the detection of GSH.This
Imply AuNCs@Tf-Cu2+To the selective enumeration method of GSH.
Embodiment 8
The research of cytotoxicity experiment.
Human cervical carcinoma cell (HeLa) (10%FBS/DMEM culture medium, 5%CO of logarithmic growth phase will be in2, 37 DEG C)
By every hole 5 × 103It is a to be inoculated in 96 well culture plates, after cell is adherent, change 200 μ L@containing AuNCs Tf (concentration is descending are as follows:
0 μM, 50 μM, 100 μM, 200 μM, 400 μM, 600 μM) culture medium prepare each experimental group, culture for 24 hours after, AuNCs@will be contained
The culture solution of Tf removes, and every hole is separately added into 200 μ L fresh cultures, and then the MTT solution of 20 μ l 5mg/ml is added in every hole,
Continue to cultivate 4h, then remove old culture solution in hole, then 150 μ LDMSO are added in every hole, shake 10min, enterprising in microplate reader
Row MTT detection.
Fig. 5 is various concentration AuNCs@Tf on human cervical carcinoma cell (HeLa) active influence.As seen from the figure, even if cell
In the case where the AuNCs@Tf of high concentration is incubated for, survival rate shows that material has relatively low toxicity still up to 80% or more.
Embodiment 9
The distribution of AuNCs@Tf in the cell.
In order to further determine the position of AuNCs@Tf in cell, green lysosome probe is used.By HeLa cell with 1.0
× 105The density of a cell is inoculated into 35mm culture dish, after culture 20 hours, AuNCs@Tf (1mg/mL) is added and is cultivated
In ware.After 1 hour, 3 times are washed to remove free AuNCs@Tf with PBS.Then green lysosome probe is added in cell
And continue after being incubated for 30 minutes, 3 times are washed to remove free lysosome probe.Finally, using laser confocal microscope
Observe cell.AuNCs@Tf is excited at 405nm, and transmitting is collected at 600-700nm.Green lysosome probe exists
488nm excitation, and collect and emit in 500-600nm.
As shown in Figure 6A, it can be observed that AuNCs@Tf issues red fluorescence, LysoTracker issues green fluorescence, and
And composograph is observed that fluorescent orange, shows AuNCs@Tf and lysosome positioning.In general, when common location coefficient is close or
When greater than 0.5 (r >=0.5), common location index is good.Therefore, it can be said that one can consider that AuNCs@Tf has entered lysosome
(Fig. 6 B).
Embodiment 10
The identification of tumour cell.
By human cervical carcinoma cell (HeLa), human liver cancer cell (HepG2), human colon cancer cell (HCT116) and mouse are at fibre
Cell (3T3) is tieed up with 1.0 × 105The density of a cell is inoculated into 35mm culture dish (preceding Three Represents tumour cell, Hou Zhedai
Table normal cell), after culture 20 hours, by AuNCs@Tf-Cu2+(Off) system is added in culture dish, with glimmering under different time
Light microscope is taken pictures.In an experiment, select AuNCs@Tf (On) as control.
In order to further illustrate AuNCS@Tf-Cu2+The ability of tumor cell, by HeLa cell and 3T3 cell with number
Measure the volume ratio mixing than 1:1.After cell is adherent, by 2mg/mL AuNCs@Tf-Cu2+Be added culture dish in 1 it is small after use fluorescence
Microscope is taken pictures.
As shown in Figure 7 A, 116 cell of HeLa, HepG2 and HCT and AuNCs@Tf-Cu2+It is incubated with, is started without glimmering
Light, with the extension of time, red fluorescent is gradually appeared and brightened.Additionally, it has been found that three kinds of cancer cells in figure 7b
In, GSH be to the enhancing of the fluorescence intensity of HepG2 cell it is most bright, compared with 116 cell of HeLa and HCT, false first-rate is normal
Number enhances 2 times and 5 times (Fig. 7 C, tables 1), implies AuNCs@Tf-Cu2+System is to the sensitivity of HepG2 cell and selectivity
It is optimal.And normal cell (3T3) and AuNCs@Tf-Cu2+When being incubated with, do not observed incubation time reaches 2 hours
To fluorescence (Fig. 7 A).As control, bright fluorescence (Fig. 8 A) is also observed after AuNCs@Tf and 3T3 are cell incubation 2 hours.
This phenomenon is had also discovered in three kinds of cancer cells, and the rate of the AuNCs@Tf of HepG2 cellular uptake is in three kinds of cancer cells
In most fast (Fig. 8 B, 8C).This can explain the AuNCs@Tf-Cu that HepG2 cell absorbs2+Rate cause GSH to system quickly
Quick response.
In order to further illustrate AuNCs@Tf-Cu2+To the selectivity of GSH, we are by HeLa cell and 3T3 cell with quantity
Ratio than 1:1 is mixed into culture dish.After cell is adherent, by AuNCs@Tf-Cu2+It is added in culture dish.After 1 hour, observation
Cell.As illustrated in fig. 7d, there is red fluorescence in the cancer cell marked in circle, and be not observed in normal cell glimmering
Light.This it is interesting the result shows that, gold nanoclusters material can target lysosome in situ restore cancer cell fluorescence, identification cancer
It has broad prospects in terms of cell, can be applied to the diagnosis and prevention of cancer.
Table 1
Embodiment 11
The research that AuNCs Tf is encrypted and decrypted as information.
AuNCs@Tf storing liquid is filled up in pen the writing of Chinese characters on filter paper by us, is dried in air.Then it uses
The ultraviolet light irradiation of 365nm.Then we are by Cu2+Solution drop on Chinese character and with ultraviolet light irradiation it.Finally, GSH solution is dripped
On Chinese character and with ultraviolet light irradiation.
We dip AuNCs Tf and by fingerprint printing and dyeing on filter paper in finger, dry in air.Then 365nm is used
Ultraviolet light irradiation fingerprint.Then we are by Cu2+Solution drop on fingerprint and with ultraviolet light irradiation it.Finally, GSH solution drop is existed
On fingerprint and with ultraviolet light irradiation.
Based on Cu2+With GSH to the stimuli responsive of fluorescence signal, we devise a kind of novel information encryption and decryption side
Case.We can be understood with AuNCs@Tf writing of Chinese characters and printing fingerprint (Fig. 9) when the UV light time for being exposed to 365nm on filter paper
See Chinese character and finger print information in ground.By Cu2+After being loaded on the filter paper for being impregnated with AuNCs@Tf, the fluorescence of Chinese character and fingerprint is quenched
It goes out, we have encrypted information in this way.Then, GSH is loaded into dipping AuNCs@Tf-Cu by us2+Chinese character and fingerprint on.Quenching
Fluorescence restore, we can reacquire information.This phenomenon shows AuNCs@Tf in information encryption and decryption processes
There is very big application prospect.
Claims (7)
1. the preparation method that a kind of transferrins is the red fluorescence gold nano cluster of template, which is characterized in that including walking as follows
It is rapid:
HAuCl is respectively configured4With the aqueous solution of Tf, 20-60:1 mixes them in molar ratio, is then stirred at room temperature 2-5 points
Solution after the pH of mixed solution is adjusted to 12 with NaOH solution, is placed in microwave reactor and is sealed, at 70-90 DEG C by clock
Reaction at least 50min obtains the AuNCs@Tf of intense red fluorescent emission.
2. a kind of transferrins as described in claim 1 is the preparation method of the red fluorescence gold nano cluster of template, special
Sign is, the HAuCl4Molar ratio with Tf is 40:1.
3. a kind of transferrins as described in claim 1 is the preparation method of the red fluorescence gold nano cluster of template, special
Sign is that reaction condition is 60min at 80 DEG C in the microwave reactor.
4. the red fluorescence gold nano cluster that method as claimed in claim 1,2 or 3 is prepared.
5. application of the red fluorescence gold nano cluster as claimed in claim 4 in detection copper ion.
6. red fluorescence gold nano cluster as claimed in claim 4 identifies cancer cell in preparation detection GSH and targeting lysosome
Application in reagent.
7. application of the red fluorescence gold nano cluster as claimed in claim 4 in the encryption and decryption of finger print information.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811580659.3A CN109705841B (en) | 2018-12-24 | 2018-12-24 | Gold nanocluster with transferrin as template and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811580659.3A CN109705841B (en) | 2018-12-24 | 2018-12-24 | Gold nanocluster with transferrin as template and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109705841A true CN109705841A (en) | 2019-05-03 |
| CN109705841B CN109705841B (en) | 2021-07-02 |
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| CN113640266B (en) * | 2021-08-11 | 2024-02-27 | 郑州大学 | Detection method for storing and releasing iron from ferritin in cells |
| CN114907840A (en) * | 2022-03-10 | 2022-08-16 | 安徽大学 | Fluorescent gold nanocluster for monitoring intracellular GSH concentration and preparation method thereof |
| CN114907840B (en) * | 2022-03-10 | 2023-05-26 | 安徽大学 | Fluorescent gold nanocluster for monitoring intracellular GSH concentration and preparation method thereof |
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