CN103880931B - Many gold bunch-apoferritin complex and preparation method thereof - Google Patents
Many gold bunch-apoferritin complex and preparation method thereof Download PDFInfo
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- CN103880931B CN103880931B CN201410103174.0A CN201410103174A CN103880931B CN 103880931 B CN103880931 B CN 103880931B CN 201410103174 A CN201410103174 A CN 201410103174A CN 103880931 B CN103880931 B CN 103880931B
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- 229910052737 gold Inorganic materials 0.000 title claims abstract description 130
- 239000010931 gold Substances 0.000 title claims abstract description 129
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 128
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 238000010668 complexation reaction Methods 0.000 title description 2
- 102000000546 Apoferritins Human genes 0.000 claims abstract description 54
- 108010002084 Apoferritins Proteins 0.000 claims abstract description 54
- 238000000034 method Methods 0.000 claims abstract description 25
- SJUCACGNNJFHLB-UHFFFAOYSA-N O=C1N[ClH](=O)NC2=C1NC(=O)N2 Chemical compound O=C1N[ClH](=O)NC2=C1NC(=O)N2 SJUCACGNNJFHLB-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000000108 ultra-filtration Methods 0.000 claims description 14
- 238000004140 cleaning Methods 0.000 claims description 11
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims description 9
- 102000004316 Oxidoreductases Human genes 0.000 claims description 6
- 108090000854 Oxidoreductases Proteins 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- 102000002067 Protein Subunits Human genes 0.000 abstract description 9
- 108010001267 Protein Subunits Proteins 0.000 abstract description 9
- 239000000243 solution Substances 0.000 description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
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- 241000894006 Bacteria Species 0.000 description 1
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- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
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- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
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- 108090000765 processed proteins & peptides Proteins 0.000 description 1
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- 102000004169 proteins and genes Human genes 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/58—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing copper, silver or gold
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- Gastroenterology & Hepatology (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Medicinal Chemistry (AREA)
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Abstract
The invention discloses the many gold bunch of one-apoferritin complex, containing the full weight chain apoferritin being made up of 24 protein subunits and the gold nano cluster being grown on described full weight chain apoferritin in these many gold bunch-apoferritin complex。The preparation method that the invention also discloses a kind of many gold bunch-apoferritin complex, the method includes contacting the full weight chain apoferritin being made up of 24 protein subunits with chlorauric acid solution。Many advantages that gold bunch-apoferritin complex can have good biocompatibility simultaneously, near-infrared fluorescent characteristic is strong, purity is high and the load factor of gold nano cluster is high provided by the invention, and the preparation method of many gold bunch-apoferritin complex provided by the invention is simple to operate。
Description
Technical field
The present invention relates to fluorescent nano material field, in particular it relates to the preparation method of a kind of many gold bunch-apoferritin complex and many gold bunch-apoferritin complex。
Background technology
Noble metal nano cluster (NMNCs) generally refers to former molecular had fluorescence, water miscible molecular level aggregation by the several of the noble metals such as Au, Ag or Pt to tens。The distinctive quantum size effect of NMNCs makes its optical property have the characteristic changed with size, and this characteristic makes the fluorescence emission spectrum of NMNCs scalable within the scope of visible ray to near-infrared region。The metals such as Au, Ag or Pt have chemical inertness and blocking group, and the toxic and side effects of organism is little so that the NMNCs containing this type of noble metal has good biocompatibility。The particle diameter of NMNCs is typically in below 2nm, and boundary is between atom and nano-particle。Additionally, NMNCs also has that light stability polymerizing power strong, anti-is strong, stoke shift is relatively big and the feature such as flicker free on all relevant time scale, substantially improve its performance as biomarker, be obviously improved the sensitivity of existing analysis method。
For over ten years, the synthetic method of NMNCs has been done very big improvement and development by researchers both domestic and external。The synthetic method of current NMNCs has multiple classification, can be divided into template and monolayer Protection Code two kinds according to the difference of blocking group。Template is to synthesize with certain material for substrate or model have special stereochemical structure or have the method for NMNCs of specific function, is one of current most common method；Monolayer protection nanocluster (MPCs) refers to the nanocluster with specific function formed in finishing or one layer of molecule of assembling。
In recent years, research for NMNCs is day by day ripe, NMNCs is greatly improved in size, chemistry and light stability, the center of gravity of research also begins to shift to various application by simply synthesizing, as being applied to biosensor, cell marking, the detection of cysteine, the detection of protein, colibacillary detection, bioprobe etc.。
Owing to the noble metal nano cluster being template with biomaterial has that biological safety is good, stability strong, can carry out the advantages such as bio-modification transformation by genetic engineering and is subject to the extensive concern of researcheres。And relate to the bioactive substance of various cell functions in organism with the biomaterial in the noble metal nano cluster that biomaterial is template, and there is good biocompatibility and controlled biological degradability, small peptide and aminoacid after degraded can also be absorbed by the body。Accordingly, with respect to other nano material systems, biomaterial has more wide application prospect。
But, the growth carrying out noble metal cluster in existing biomaterial also has complicated operation, near-infrared fluorescent characteristic is weak, biocompatibility is bad, can not accurately control the defect such as growth of multiple nano-noble metal clusters in biomaterial, remains an insoluble problem。
Summary of the invention
It is an object of the invention to overcome the defect of prior art, it is provided that a kind of preparation method is easy, near-infrared fluorescent penetration capacity is strong and can accurately control many gold bunch of the growth of multiple nano-noble metal clusters-apoferritin complex in biomaterial。
To achieve these goals, the invention provides the many gold bunch of one-apoferritin complex, containing the full weight chain apoferritin being made up of 24 protein subunits and the gold nano cluster being grown on described full weight chain apoferritin in these many gold bunch-apoferritin complex。
The preparation method that present invention also offers a kind of many gold bunch-apoferritin complex, the method includes contacting the full weight chain apoferritin being made up of 24 protein subunits with chlorauric acid solution。
Present invention also offers a kind of many gold bunch-apoferritin complex prepared according to the preparation method of the present invention。
Many advantages that gold bunch-apoferritin complex can have good biocompatibility simultaneously, near-infrared fluorescent characteristic is strong, purity is high and the load factor of gold nano cluster is high provided by the invention, and the preparation method of many gold bunch-apoferritin complex provided by the invention is simple to operate。
Other features and advantages of the present invention will be described in detail in detailed description of the invention part subsequently。
Accompanying drawing explanation
Accompanying drawing is used to provide a further understanding of the present invention, and constitutes the part of description, is used for explaining the present invention, but is not intended that limitation of the present invention together with detailed description below。In the accompanying drawings:
Fig. 1 represents the shape appearance figure being observed many gold bunch-apoferritin complex obtained in the embodiment of the present invention 1 by Ice mapping。
Fig. 2 represents by the high-resolution imaging of gold nano cluster in many gold bunch obtained in the transmission electron microscope observing embodiment of the present invention 1-apoferritin complex。
Fig. 3 represents in many gold bunch-apoferritin complex obtained in the embodiment of the present invention 1 the X-radial energy spectrogram of gold atom in gold nano cluster。
Fig. 4 represents the fluorescence excitation of many gold bunch-apoferritin complex obtained in the embodiment of the present invention 1 and launches collection of illustrative plates。
Detailed description of the invention
Hereinafter the specific embodiment of the present invention is described in detail。It should be appreciated that detailed description of the invention described herein is merely to illustrate and explains the present invention, it is not limited to the present invention。
In the present invention, when not making contrary explanation, the scope of the term " solution " used is not limited to the particle diameter of the dispersate disperse system (true solution) less than 1nm, but refers to homogeneous liquefied mixture, it is possible to include colloidal dispersion (colloid solution)。
In the present invention, when not making contrary explanation, the volumetric quantities of liquid is the numerical value under standard state。
In the present invention, when not making contrary explanation, described contact or mixing can carry out when stirring。The speed of stirring can be conventional selection。
In the present invention, described " full weight chain apoferritin " is the nonferrous ferritin being made up of 24 heavy chains (H-chain), and its serial number is: RCSBProteinDataBank RCSBPDB:3AJO。
The invention provides the many gold bunch of one-apoferritin complex, containing the full weight chain apoferritin being made up of 24 protein subunits and the gold nano cluster being grown on described full weight chain apoferritin in these many gold bunch-apoferritin complex。
In the present invention, described full weight chain apoferritin can pass through conventional use of various methods in this area and obtain, such as can extract from escherichia coli and obtain, in the present invention, it is preferred to use document (MasakiUchida etc., TargetingofCancerCellswithFerrimagneticFerritinCageNanop articles, JournalofAmericanChemicalSociety, 2006, Vol28,16626-16633) method that provides obtains full weight chain apoferritin。
In many gold bunch-apoferritin complex of the present invention, it is preferable that the protein subunit of described full weight chain apoferritin has the ferrous oxidase site containing histidine。
In many gold bunch-apoferritin complex of the present invention, it is preferable that described gold nano cluster is positioned on the ferrous oxidase site of full weight chain apoferritin, or described gold nano cluster is positioned at around the ferrous oxidase site of full weight chain apoferritin。
By above-mentioned many advantages that gold bunch-apoferritin complex has good biocompatibility, near-infrared fluorescent characteristic is strong provided by the invention。
The preparation method that present invention also offers a kind of many gold bunch-apoferritin complex, the method includes contacting the full weight chain apoferritin being made up of 24 protein subunits with chlorauric acid solution。
In the present invention, described full weight chain apoferritin preferably provides in the form of a solution, and solvent is preferably water, and more preferably the concentration of full weight chain apoferritin aqueous solution is 1-20mg/mL。
The inventive point of the present invention essentially consists in and is contacted and/or mix many gold bunch-apoferritin complex of obtaining and preparation method thereof by above-mentioned full weight chain apoferritin with chlorauric acid solution, present invention only requires and full weight chain apoferritin is contacted with chlorauric acid solution and/or mixes the many gold bunch-apoferritin complex that can obtain there is good biocompatibility, near-infrared fluorescent characteristic is strong, purity is high and the load factor of gold nano cluster is high, as can be seen here, the preparation method operation of many gold bunch-apoferritin complex provided by the invention is very simple。
In the preparation method of many gold bunch-apoferritin complex of the present invention, the full weight chain apoferritin of relatively every weight portion, it is preferable that the consumption of described gold chloride is 0.5-4 weight portion。
In the preparation method of many gold bunch-apoferritin complex of the present invention, the full weight chain apoferritin of relatively every weight portion, it is preferred that the consumption of gold chloride is 1-2 weight portion。
In the preparation method of many gold bunch-apoferritin complex of the present invention, it is preferable that the pH value of described contact is 11-13；It is preferred that pH value is 11.5-12.5。
In the preparation method of many gold bunch-apoferritin complex of the present invention, it is preferable that the temperature of described contact is 4-45 DEG C；It is preferred that temperature is 15-37 DEG C。
In the preparation method of many gold bunch-apoferritin complex of the present invention, it is preferable that the time of described contact is 12-48 hour；It is preferred that the time is 20-35 hour。
In the preparation method of many gold bunch-apoferritin complex of the present invention, it is preferable that the concentration of described chlorauric acid solution is 10-50 mM/l；It is preferred that the concentration of described chlorauric acid solution is 22-38 mM/l。
In the preparation method of many gold bunch-apoferritin complex of the present invention, it is preferable that the method also includes the mixture after by contact and is sequentially carried out centrifugal, ultrafiltration and cleaning, obtains many gold bunch-apoferritin complex。
In method of the present invention, to the method for described centrifugal, ultrafiltration and cleaning and condition, there is no particular limitation, it is possible to adopts conventional use of various methods in this area。It is for instance possible to use centrifuge is centrifuged under the rotating speed of 5000-15000 rev/min, then centrifugal rear obtained supernatant is sequentially carried out ultrafiltration and cleaning。In the present invention, the method for described ultrafiltration has multiple, present invention preferably employs and by super filter tube and/or ultrafilter membrane, centrifugal rear obtained supernatant is carried out ultrafiltration。The present invention preferably takes appropriate deionized water and filtrate obtained after ultrafiltration is carried out。
A preferred embodiment of the invention, the preparation method of described many gold bunch-apoferritin complex includes contacting the full weight chain apoferritin being made up of 24 protein subunits with chlorauric acid solution, the full weight chain apoferritin of relatively every weight portion, the consumption of gold chloride is 0.5-4 weight portion, it is 11-13 that the condition of described contact includes pH value, temperature is 4-45 DEG C, and the time is 12-48 hour, and the concentration of described chlorauric acid solution is 10-50 mM/l。Then many gold bunch-apoferritin complex obtained after being contacted with chlorauric acid solution by the full weight chain apoferritin being made up of 24 protein subunits is sequentially carried out centrifugal, ultrafiltration and cleaning。
In the present invention, in case of no particular description, the solvent in described " solution " is water。Such as, described chlorauric acid solution is aqueous solution of chloraurate。
Hereinafter, the present invention is further described by embodiment。Wherein, in case of no particular description, reagent used is commercially available product。
In following example of the present invention, No. CAS of the gold chloride used is 16961-25-4, purchased from sigma company；
The deionized water used is State Nanometer Science Center's bio-safety and the homemade deionized water in effect experiment room；
The centrifuge trade mark used is XIR, purchased from Thermo company；
The 50KD super filter tube article No. UFC801096 used, purchased from MIlipore company；
The Ice mapping FEITitanKrios used, purchased from FEI Co.；
The high-resolution-ration transmission electric-lens FEITecnaiF20U-TWIN used, purchased from FEI Co.；
The model FEITecnaiF20U-TWIN of the X-ray energy spectrum measuring instrument used, is purchased from FEI Co.；
The model of the spectrofluorophotometer used is Hitachi F-4600, is purchased from HITACHI company。
Preparation example 1
This prepares example for obtaining full weight chain apoferritin of the present invention。
Adopt document (MasakiUchida etc., TargetingofCancerCellswithFerrimagneticFerritinCageNanop articles, JournalofAmericanChemicalSociety, 2006, Vol28,16626-16633) method that provides obtains full weight chain apoferritin, institute the difference is that, prepare in example at this and expression strain of full weight chain apoferritin is cultured to OD=0.6-0.8 in 37 DEG C, add IPTG abduction delivering 4 hours at 37 DEG C of final concentration of 0.5mM。Every liter of bacterium obtains the albumen of purification at more than 50mg, and purity is 97.2%。
The concentration of full weight chain apoferritin solution is determined by the absorbance at 280nm place, and its concentration is 10mg/mL, extracts 100mg altogether, and purity is 97.2%。
Preparation example 2
This prepares example for obtaining chlorauric acid solution of the present invention。
Take the gold chloride of 5 parts of 1g/ parts, it is dissolved in the deionized water of 60mL, 45mL, 50mL, 75mL and 120mL respectively, then adjust pH value to 7 with the sodium hydroxide solution that concentration is 0.1 mol/L, respectively obtain the chlorauric acid solution of 30 mM/ls, 45 mM/ls, 38 mM/ls, 22 mM/ls and 12 mM/ls, standby。
Embodiment 1
The present embodiment is used for many gold bunch-apoferritin complex A of the present invention and preparation method thereof are described。
Take the chlorauric acid solution 3.025mL of 30 mM/ls that prepare example 2 gained, join in the 5mL full weight chain apoferritin aqueous solution preparing example 1 gained 5mg/mL。Adjusting PH to 12 with the sodium hydroxide solution that concentration is 0.1 mol/L, in the constant incubator of 37 DEG C, oscillation incubation is after 24 hours, obtained solution proceeds to the centrifugation 10min with 7500 revs/min in centrifuge, takes supernatant。Then obtained supernatant is carried out ultrafiltration by 50KD super filter tube, solution obtained after cleaning ultrafiltration by appropriate amount of deionized water again, repeat this cleaning step 3 times, with remove solution is likely to containing gold chloride, obtain many gold bunch-apoferritin complex A that purity is 97.5%。
By Ice mapping it can be seen that there is multiple gold nano cluster the inside of the above-mentioned many gold bunch-apoferritin complex A prepared, as shown in Figure 1。
By high-resolution-ration transmission electric-lens it can be seen that the lattice structure of the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex A prepared, as shown in Figure 2。
By X-ray energy spectrum measuring instrument it can be seen that the X-ray energy spectrum of gold element in the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex A prepared, as shown in Figure 3。
Embodiment 2
The present embodiment is used for many gold bunch-apoferritin complex B of the present invention and preparation method thereof are described。
Take the chlorauric acid solution 2.758mL of 22 mM/ls that prepare example 2 gained, join in the 5mL full weight chain apoferritin aqueous solution preparing example 1 gained 5mg/mL。Adjusting PH to 11.5 with sodium hydroxide, in the water bath of 30 DEG C of constant temperature, oscillation incubation is after 20 hours, obtained solution proceeds to the centrifugation 10min with 10000 revs/min in centrifuge, takes supernatant。Then obtained supernatant is carried out ultrafiltration by 50KD super filter tube, solution obtained after cleaning ultrafiltration by appropriate amount of deionized water again, repeat this cleaning step 3 times, with remove solution is likely to containing gold chloride, obtain many gold bunch-apoferritin complex B that purity is 97.3%。
By Ice mapping it can be seen that there is multiple gold nano cluster the inside of the above-mentioned many gold bunch-apoferritin complex B prepared, with how gold bunch-complex A is similar for apoferritin。
By high-resolution-ration transmission electric-lens it can be seen that the lattice structure of the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex B prepared, with many gold bunch-complex A is similar for apoferritin。
By X-ray energy spectrum measuring instrument it can be seen that the X-ray energy spectrum of gold element in the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex B prepared, with many gold bunch-complex A is similar for apoferritin。
Embodiment 3
The present embodiment is used for many gold bunch-apoferritin complex C of the present invention and preparation method thereof are described。
Take the chlorauric acid solution 3.194mL of 38 mM/ls that prepare example 2 gained, join in the 5mL full weight chain apoferritin aqueous solution preparing example 1 gained 5mg/mL。Adjusting PH to 12.5 with sodium hydroxide, in the water bath of 30 DEG C of constant temperature, oscillation incubation is after 35 hours, obtained solution proceeds to the centrifugation 10min with 8000 revs/min in centrifuge, takes supernatant。Then obtained supernatant is carried out ultrafiltration by 50KD super filter tube, solution obtained after cleaning ultrafiltration by appropriate amount of deionized water again, repeat this cleaning step 3 times, with remove solution is likely to containing gold chloride, obtain many gold bunch-apoferritin complex C that purity is 97.6%。
By Ice mapping it can be seen that there is multiple gold nano cluster the inside of the above-mentioned many gold bunch-apoferritin complex C prepared, with how gold bunch-complex A is similar for apoferritin。
By high-resolution-ration transmission electric-lens it can be seen that the lattice structure of the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex C prepared, with many gold bunch-complex A is similar for apoferritin。
By X-ray energy spectrum measuring instrument it can be seen that the X-ray energy spectrum of gold element in the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex C prepared, with many gold bunch-complex A is similar for apoferritin。
Embodiment 4
The present embodiment is used for many gold bunch-apoferritin complex D of the present invention and preparation method thereof are described。
The present embodiment adopts and prepares many gold bunch-apoferritin complex D with the identical method of embodiment 1, institute the difference is that:
The chlorauric acid solution 0.674mL of take preparation example 2 gained 45 mM/ls joins haptoreaction in full weight chain apoferritin aqueous solution。
The purity of the many gold bunch obtained-apoferritin complex D is 95.0%。
By Ice mapping it can be seen that there is multiple gold nano cluster the inside of the above-mentioned many gold bunch-apoferritin complex D prepared, but it can be seen that there is a small amount of red impurity in the solution of the many gold bunch-apoferritin complex D of yellow。
By high-resolution-ration transmission electric-lens it can be seen that the lattice structure of the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex D prepared, with many gold bunch-complex A is similar for apoferritin。
By X-ray energy spectrum measuring instrument it can be seen that the X-ray energy spectrum of gold element in the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex D prepared, with many gold bunch-complex A is similar for apoferritin。
Embodiment 5
The present embodiment is used for many gold bunch-apoferritin complex E of the present invention and preparation method thereof are described。
The present embodiment adopts and prepares many gold bunch-apoferritin complex E with the identical method of embodiment 1, institute the difference is that:
The chlorauric acid solution 20.227mL of take preparation example 2 gained 12 mM/ls joins haptoreaction in full weight chain apoferritin aqueous solution。
The purity of the many gold bunch obtained-apoferritin complex E is 96.5%。
By Ice mapping it can be seen that there is multiple gold nano cluster the inside of the above-mentioned many gold bunch-apoferritin complex E prepared, but it can be seen that there is a small amount of red impurity in the solution of the many gold bunch-apoferritin complex E of yellow。
By high-resolution-ration transmission electric-lens it can be seen that the lattice structure of the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex E prepared, with many gold bunch-complex A is similar for apoferritin。
By X-ray energy spectrum measuring instrument it can be seen that the X-ray energy spectrum of gold element in the gold nano cluster of the inside of the above-mentioned many gold bunch-apoferritin complex E prepared, with many gold bunch-complex A is similar for apoferritin。
Test case 1
This test case adopts many gold bunch-apoferritin complex A(Au-HFt of being prepared by above-described embodiment 1 of fluorescent spectrophotometer assay) near-infrared fluorescent penetration capacity, its fluorescence excitation and launch collection of illustrative plates as shown in Figure 4。
The near-infrared fluorescent penetration capacity of many gold bunch-apoferritin complex A that the method that as can be seen from Figure 4 employing present example 1 provides prepares is strong, and fluorescent characteristic is good, and its fluorescence excitation peak is at about 720nm, and emission peak is at about 810nm。
Test case 2-5
The mode identical with test case 1 is adopted to measure the many gold bunch-apoferritin complex B prepared by above-described embodiment 2-5, many gold bunch-apoferritin complex C, many gold bunch-apoferritin complex D, many gold bunch-apoferritin complex E, record the near-infrared fluorescent penetration capacity of above-mentioned many gold bunch-apoferritin complex, its fluorescence excitation and transmitting collection of illustrative plates and how gold bunch-complex A is similar for apoferritin。
By detecting above and analysis is appreciated that, many gold bunch-purity height of apoferritin complex provided by the invention, near-infrared fluorescent penetration capacity, gold nano cluster load factor height, and the method preparing many gold bunch-apoferritin complex provided by the invention is easy and simple to handle。
The preferred embodiment of the present invention described in detail above; but, the present invention is not limited to the detail in above-mentioned embodiment, in the technology concept of the present invention; technical scheme can being carried out multiple simple variant, these simple variant belong to protection scope of the present invention。
It is further to note that each the concrete technical characteristic described in above-mentioned detailed description of the invention, in reconcilable situation, it is possible to be combined by any suitable mode, various possible compound modes are no longer illustrated by the present invention separately。
Additionally, can also carry out combination in any between the various different embodiment of the present invention, as long as it is without prejudice to the thought of the present invention, it should be considered as content disclosed in this invention equally。
Claims (8)
1. gold bunch-apoferritin complex more than a kind, it is characterized in that, containing the people source full weight chain apoferritin being made up of 24 people source apoferritin heavy chain subunit and the gold nano cluster being grown on the full weight chain apoferritin of described people source in these many gold bunch-apoferritin complex, wherein, the heavy chain subunit of described people source full weight chain apoferritin has the ferrous oxidase site containing histidine, described gold nano cluster is positioned on the ferrous oxidase site of people source full weight chain apoferritin, or described gold nano cluster is positioned at around the ferrous oxidase site of people source full weight chain apoferritin。
2. the preparation method of gold bunch-apoferritin complex more than a kind, it is characterized in that, the method includes contacting the people source full weight chain apoferritin being made up of 24 people source apoferritin heavy chain subunit with chlorauric acid solution, the people source full weight chain apoferritin of relatively every weight portion, the consumption of described gold chloride is 1-2 weight portion。
3. method according to claim 2, wherein, it is 11-13 that the condition of described contact includes pH value, and temperature is 4-45 DEG C, and the time is 12-48 hour。
4. method according to claim 3, wherein, it is 11.5-12.5 that the condition of described contact includes pH value, and temperature is 15-37 DEG C, and the time is 20-35 hour。
5. the method according to any one in claim 2-4, wherein, the concentration of described chlorauric acid solution is 10-50 mM/l。
6. method according to claim 5, wherein, the concentration of described chlorauric acid solution is 22-38 mM/l。
7. method according to claim 5, wherein, the method also includes the mixture after by contact and is sequentially carried out centrifugal, ultrafiltration and cleaning, obtains many gold bunch-apoferritin complex。
8. many gold bunch-apoferritin complex that in claim 2-7, method described in any one obtains。
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