CN110006863A - Clinical medicine analysis method based on human albumin's functionalized nano silica - Google Patents
Clinical medicine analysis method based on human albumin's functionalized nano silica Download PDFInfo
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Abstract
The invention belongs to biomedicine technical fields, disclose a kind of clinical medicine analysis method based on human albumin's functionalized nano silica, prepare silica, by nano-silica particle surface modification APTS and surface amidation；Carry out the fluorescence lifetime analysis based on human albumin's silica；Carry out the characterization of molecules discriminance analysis based on human albumin's nano silica；It carries out based on human albumin's nano silica detection analysis；Since silica is because having the characteristics that ordered mesopore structure, large specific surface area, good biocompatibility and surface are easy to modify, great application prospect is shown in fields such as biological medicines, the space arrangement of the specific shape of identification molecule of serum protein molecule abundance, size and its chemical functional base is analyzed, to the Selective long-range DEPT of haemocyanin, molecularly imprinted polymer after analysis substantially has specificity height, and preparation is simple, stability is good, reusable.
Description
Technical field
The invention belongs to biomedicine technical fields, more particularly to one kind to be based on human albumin's functionalized nano titanium dioxide
The clinical medicine analysis method of silicon.
Background technique
Currently, the prior art commonly used in the trade is such that nano silica, it is in tridimensional network, surface exists
The hydroxyl of a large amount of unsaturation residual bond and different conditions, this enables nano-silica surface high, unstable in thermodynamics
State has very high chemical activity.Wherein since mesoporous silicon oxide is because having ordered mesopore structure, large specific surface area, life
Object compatibility is good and surface is easy to the features such as modifying, and shows great application prospect in fields such as biological medicines.Current two
Silica identifies that the insufficient space to the specific shape of molecule, size and its chemical functional base is arranged to serum protein molecule
Column are analyzed, poor to the selectivity of haemocyanin；Silica analyzes deficiency to serum protein molecule, although identification is special
One property is very high, but preparation is complicated, stability is poor, is easy the binding site that loses activity.
In conclusion problem of the existing technology is:
(1) at present silica to serum protein molecule identify it is insufficient to the specific shape of molecule, size and its
The space arrangement of chemical functional base is analyzed, poor to the selectivity of haemocyanin.
(2) silica is analyzed serum protein molecule insufficient, although the specificity of identification is very high, preparation is multiple
It is miscellaneous, stability is poor, is easy the binding site that loses activity.
Summary of the invention
In view of the problems of the existing technology, the present invention provides based on human albumin's functionalized nano silica
Clinical medicine analysis method.
The invention is realized in this way a kind of clinical medicine analysis based on human albumin's functionalized nano silica
Method, comprising the following steps:
Step 1, using TEOS as predecessor, with NH3As catalyst, ethyl alcohol prepares silica as solvent；
Step 2, by nano-silica particle surface modification APTS and surface amidation；
Step 3 carries out the fluorescence lifetime analysis based on human albumin's silica；
Step 4 carries out the characterization of molecules discriminance analysis based on human albumin's nano silica；
Step 5 is carried out based on human albumin's nano silica detection analysis.
Further, in step 3, the fluorescence lifetime analysis based on human albumin's nano silica are as follows:
Decay time histogram is acquired between fluorescent emission and excitation using microchannel pulse-height analyzer, is then turned
Become data-signal to go to handle this process to computer, instantaneous emission spectrum is to pass through use to the measurement of the receptance function of time
FITC-APTS-silica particle suspension, which is dispersed in the haemocyanin target analysis of various concentration, to be obtained.
Further, in step 4, the characterization of molecules discriminance analysis based on human albumin's nano silica are as follows:
(1) it is analyzed in ethanol/acetonitrile solution between template molecule and propylene phthalein amine molecule by ultraviolet visible spectrometry
Active force；
(2) the haemocyanin solution that various concentration is added dropwise observes ultraviolet-visible light spectral curve.
Further, provided in an embodiment of the present invention to be based on human albumin's nano silica detection analysis in step 5
Are as follows:
(1) FITC-APTS-silica particle is added in fluorescence spectrum quartz colorimetric utensil；
(2) oneself is known that the haemocyanin solution of concentration is injected into cuvette to obtain steady-state fluorescence spectrum, is based on
The detection analysis of haemocyanin.
Further, the step 4 is based in the characterization of molecules discriminance analysis of human albumin's nano silica, into one
Step includes: to choose the characteristic parameter CP of several characterization of molecules interference signals based on human albumin's nano silica, packet
Include frequency F, time T, for observation point airspace angle Θ, polarization direction Γ and coding mode C, and using parameter as coordinate
Axis establishes multidimensional coordinate system, for each reference axis of coordinate system, respectively according to the resolution ratio of each interference characteristic parameter
The unit quantity for determining respective coordinates axis is defined as interference space based on the spatial model of establishment of coordinate system multidimensional characteristic parameter:
HSI=SPACE (CP1,CP2…CPN), any one vector in this interference spaceIt is indicated by coordinate:
Wherein CPiFor vectorThe coordinate of certain dimension in interference space is for characteristic parameter CPiSpecific value,
For the interference signal of single mode, a single vector is expressed as in interference space:
For the interference signal of multimode, i.e. the one of interference signal or several characteristic parameters shows as with multivalue spy
Property, the set expression of interference characteristic vector is used in interference space:
Another object of the present invention is to provide a kind of clinic based on human albumin's functionalized nano silica
Pharmaceutical Analysis system, comprising:
Nano silica prepares module, nano-silica surface processing module, the fluorescence lifetime based on haemocyanin
Analysis module, the characterization of molecules discriminance analysis module based on haemocyanin, the detection analysis module based on haemocyanin；
Nano silica preparation module is connect with nano-silica surface processing module, using TEOS as predecessor, with
NH3 prepares silica as solvent as catalyst, ethyl alcohol；
Fluorescence lifetime analysis module based on haemocyanin is connect with nano-silica surface processing module, for carrying out
Fluorescence lifetime analysis based on human albumin's nano silica；
Characterization of molecules discriminance analysis module based on haemocyanin is connect with nano-silica surface processing module, is used for
Carry out the characterization of molecules discriminance analysis based on human albumin's nano silica；
Detection analysis module based on haemocyanin is connect with nano-silica surface processing module, for being based on
Human albumin's nano silica detection analysis.
The positive effect of the present invention are as follows:
(1) the sufficient space row to the specific shape of serum protein molecule identification molecule, size and its chemical functional base
Column are analyzed, to the Selective long-range DEPT of haemocyanin.
(2) molecularly imprinted polymer after analyzing substantially has specificity height, and preparation is simple, stability is good,
It is reusable.
The characteristic parameter of the present invention using description disturbance signal establishes coordinate system as reference axis, passes through the interference space of building
Model can support the analysis, expression and concrete operation of wireless communication system interference signal using vector representation and operation, from
It and is that system forms Mathematics Proof to the judgement, analysis and management of interference signal.It, can on the basis of interference space model support
Interference management techniques are carried out as system in the method by mathematics, and guidance and help is provided；It is formed using mathematical space concept dry
Spatial model is disturbed, the state of interference signal in wireless communication system is analyzed and characterized, creative proposes various dimensions
Disturbance state spatial model.It can get accurately analysis characteristic information.
Detailed description of the invention
Fig. 1 is the clinical medicine analysis provided in an embodiment of the present invention based on human albumin's functionalized nano silica
Method flow diagram.
Fig. 2 is the clinical medicine analysis provided in an embodiment of the present invention based on human albumin's functionalized nano silica
System diagram.
In figure: 1, nano silica prepares module；2, nano-silica surface processing module；3, it is based on haemocyanin
Fluorescence lifetime analysis module；4, the characterization of molecules discriminance analysis module based on haemocyanin；5, based on the detection of haemocyanin
Analysis module.
Fig. 3 is provided in an embodiment of the present invention based on human albumin's nano silica detection analysis figure.
Specific embodiment
In order to further understand the content, features and effects of the present invention, the following examples are hereby given, and cooperate attached drawing
Detailed description are as follows.
Structure of the invention is explained in detail with reference to the accompanying drawing.
As shown in Figure 1, the clinical medicine provided in an embodiment of the present invention based on human albumin's functionalized nano silica
Object analysis method, comprising the following steps:
S101: using TEOS as predecessor, with NH3As catalyst, ethyl alcohol prepares silica as solvent.
S102: by nano-silica particle surface modification APTS and surface amidation.
S103: the fluorescence lifetime analysis based on human albumin's silica is carried out.
S104: the characterization of molecules discriminance analysis based on human albumin's nano silica is carried out.
S105: it carries out based on human albumin's nano silica detection analysis.
In step S103, the fluorescence lifetime analysis provided in an embodiment of the present invention based on human albumin's nano silica
Are as follows: decay time histogram is acquired between fluorescent emission and excitation using microchannel pulse-height analyzer, is then transformed into
Data-signal goes to handle this process to computer, and instantaneous emission spectrum is by with FITC- to the measurement of the receptance function of time
APTS-silica particle suspension, which is dispersed in the haemocyanin target analysis of various concentration, to be obtained.
In step S104, the characterization of molecules identification provided in an embodiment of the present invention based on human albumin's nano silica
Analysis are as follows:
(1) it is analyzed in ethanol/acetonitrile solution between template molecule and propylene phthalein amine molecule by ultraviolet visible spectrometry
Active force；
(2) the haemocyanin solution that various concentration is added dropwise observes ultraviolet-visible light spectral curve.
In step S104, in the characterization of molecules discriminance analysis based on human albumin's nano silica, further comprise:
Choose the characteristic parameter CP of several characterization of molecules interference signals based on human albumin's nano silica, including frequency F,
Time T, for observation point airspace angle Θ, polarization direction Γ and coding mode C, and using parameter as reference axis establish it is more
Coordinate system is tieed up, for each reference axis of coordinate system, respectively according to the resolution ratio of each interference characteristic parameterIt determines and corresponds to
The unit quantity of reference axis is defined as interference space based on the spatial model of establishment of coordinate system multidimensional characteristic parameter: HSI=SPACE
(CP1,CP2…CPN), any one vector in this interference spaceIt is indicated by coordinate:
Wherein CPiFor vectorThe coordinate of certain dimension in interference space is for characteristic parameter CPiSpecific value,
For the interference signal of single mode, a single vector is expressed as in interference space:
For the interference signal of multimode, i.e. the one of interference signal or several characteristic parameters shows as with multivalue spy
Property, the set expression of interference characteristic vector is used in interference space:
It needs to be determined that interference signal and contrast signal are in wireless communication before several interference signal characteristic parameters of selection
Physical parameter on number field, frequency F, time T including wireless signal, for observation point airspace angle Θ, polarization direction Γ,
And coding mode C.
Characteristic parameter is used to establish space coordinates, the interference space of definition as reference axis are as follows:
HSI=SPACE (F, T, Θ, Γ, C)；
Any one vector in interference spaceIt is indicated by the coordinate in interference characteristic space:
F therein, t, θ, γ, c correspond to the coordinate value of each dimension respectively.
For the interference signal of single mode, a single vector is expressed as in interference space:
F therein, t, θ, γ, c correspond to the coordinate value of each dimension respectively.
Interference signal is multimode signal, i.e., the one of interference signal or several characteristic parameters show as with multivalue spy
Property, the set expression of interference characteristic vector is used in interference space:
It is a set of vectors in interference space, representative is the subspace area for meeting certain Operations of Interva Constraint condition
Domain.
It is provided in an embodiment of the present invention to be based on human albumin's nano silica detection analysis in step S105 are as follows:
(1) FITC-APTS-silica particle is added in fluorescence spectrum quartz colorimetric utensil.
(2) oneself is known that the haemocyanin solution of concentration is injected into cuvette to obtain steady-state fluorescence spectrum, is based on
The detection analysis of haemocyanin.
As shown in Fig. 2, the clinical medicine provided in an embodiment of the present invention based on human albumin's functionalized nano silica
Object analysis system, comprising:
Nano silica prepares module 1, nano-silica surface processing module 2, the fluorescence longevity based on haemocyanin
Order analysis module 3, the characterization of molecules discriminance analysis module 4 based on haemocyanin, the detection analysis module 5 based on haemocyanin.
Nano silica preparation module 1 is connect with nano-silica surface processing module 2, using TEOS as predecessor,
With NH3 as catalyst, ethyl alcohol prepares silica as solvent.
Fluorescence lifetime analysis module 3 based on haemocyanin is connect with nano-silica surface processing module 2, for into
Fluorescence lifetime analysis of the row based on human albumin's nano silica.
Characterization of molecules discriminance analysis module 4 based on haemocyanin is connect with nano-silica surface processing module 2, is used
In carrying out the characterization of molecules discriminance analysis based on human albumin's nano silica.
Detection analysis module 5 based on haemocyanin is connect with nano-silica surface processing module 2, for carrying out base
In human albumin's nano silica detection analysis.
Application principle of the invention is further described combined with specific embodiments below；
Embodiment 1: preparation nano silica
Using TEOS as predecessor, with NH3As catalyst, ethyl alcohol prepares silica as solvent.First by TEOS and
500ml is added in three cervical vertebra round-bottomed flasks of grinding port plug in the mixing of suitable ethyl alcohol, with magnetic stirring apparatus, leniently, with solid
Determine revolving speed (500rpm) stirring 3min, make to be sufficiently mixed with ethyl alcohol, then suitable deionized water and ammonium hydroxide are mixed, carries out mild
Ground is stirred with fixed rotating speed (500rpm), and ammonium hydroxide and deionized water are uniformly mixed, then the two is mixed and is rapidly added
Into above-mentioned reaction solution, rapidly, consumingly, with 700rpm stirring three minutes after, then gentle agitation (500rpm) fixed rotating speed 15
To 18h, silica dioxide gel solution can be prepared by, the addition of ammonium hydroxide must be rapid, and the volume of reaction vessel cannot be too small, in order to avoid
Dispersion unevenly cause silicon dioxide granule to agglomerate into particle excessive.By nano-silica particle surface modification APTS (ammonia
Propyl) and surface amidation.The nano silica gel particles that Centrifugal dispersion is good are taken, it is dispersed in again with dry toluene
The three neck ground round-bottomed flasks with reflux unit in, APTS is promptly added dropwise in this solution, magnetic agitation is heated to back
Thermotonus is flowed to for 24 hours by nano silica modified amino acid；By the above-mentioned obtained surface modification dioxy of aminopropyl
The reaction solution of SiClx nanoparticle is centrifuged in supercentrifuge with centrifuge tube, super in a high power numerical control ultrasonic cleaner
Sound dispersion, then respectively with dry toluene centrifugation, ultrasonic disperse three times, wash away the APTS not reacted completely, nothing finally
In single neck ground round-bottomed flask that water-toluene distributes it to, anhydrous K is added2CO3After bath ultrasound, acryloyl chloride is promptly added,
Grinding port plug is covered, reacts 12h amidation at room temperature.
Embodiment 2: the fluorescence lifetime analysis based on human albumin's nano silica
Decay time histogram is acquired between fluorescent emission and excitation with microchannel pulse-height analyzer, is then changed
Go to handle this process to computer at data-signal, instantaneous emission spectrum is by with FITC- to the measurement of the receptance function of time
APTS-silica particle suspension, which is dispersed in the haemocyanin target analysis of various concentration, to be obtained.
Embodiment 3: the characterization of molecules discriminance analysis based on human albumin's nano silica
It is analyzed in ethanol/acetonitrile solution between template molecule and propylene phthalein amine molecule by ultraviolet visible spectrometry first
Active force, the haemocyanin solution that various concentration is then added dropwise observe ultraviolet-visible light spectral curve.
Embodiment 4: it is based on human albumin's nano silica detection analysis
FITC-APTS-silica particle is added in fluorescence spectrum quartz colorimetric utensil, oneself of then handle knows the serum of concentration
Protein solution is injected into cuvette to obtain steady-state fluorescence spectrum, carries out the detection analysis based on haemocyanin.Such as Fig. 3.
In the above-described embodiments, can come wholly or partly by software, hardware, firmware or any combination thereof real
It is existing.When using entirely or partly realizing in the form of a computer program product, the computer program product include one or
Multiple computer instructions.When loading on computers or executing the computer program instructions, entirely or partly generate according to
Process described in the embodiment of the present invention or function.The computer can be general purpose computer, special purpose computer, computer network
Network or other programmable devices.The computer instruction may be stored in a computer readable storage medium, or from one
Computer readable storage medium is transmitted to another computer readable storage medium, for example, the computer instruction can be from one
A web-site, computer, server or data center pass through wired (such as coaxial cable, optical fiber, Digital Subscriber Line (DSL)
Or wireless (such as infrared, wireless, microwave etc.) mode is carried out to another web-site, computer, server or data center
Transmission).The computer-readable storage medium can be any usable medium or include one that computer can access
The data storage devices such as a or multiple usable mediums integrated server, data center.The usable medium can be magnetic Jie
Matter, (for example, floppy disk, hard disk, tape), optical medium (for example, DVD) or semiconductor medium (such as solid state hard disk Solid
State Disk (SSD)) etc..
The above is only the preferred embodiments of the present invention, and is not intended to limit the present invention in any form,
Any simple modification made to the above embodiment according to the technical essence of the invention, equivalent variations and modification, belong to
In the range of technical solution of the present invention.
Claims (10)
1. a kind of clinical medicine analysis method based on human albumin's functionalized nano silica, which is characterized in that described
The clinical medicine analysis method based on human albumin's functionalized nano silica the following steps are included:
Step 1 prepares silica: using TEOS as predecessor, with NH3As catalyst, ethyl alcohol prepares titanium dioxide as solvent
Silicon；
Nano-silica particle is surface modified APTS and surface amidation by step 2；
Step 3 carries out the fluorescence lifetime analysis based on human albumin's silica；
Step 4 carries out the characterization of molecules discriminance analysis based on human albumin's nano silica；
Step 5 is carried out based on human albumin's nano silica detection analysis.
2. the clinical medicine analysis method as described in claim 1 based on human albumin's functionalized nano silica,
It is characterized in that, in the step 3, the fluorescence lifetime analysis based on human albumin's nano silica are as follows:
Decay time histogram is acquired between fluorescent emission and excitation using microchannel pulse-height analyzer, is then transformed into
Data-signal goes to handle this process to computer, and instantaneous emission spectrum is by with FITC- to the measurement of the receptance function of time
APTS-silica particle suspension, which is dispersed in the haemocyanin target analysis of various concentration, to be obtained.
3. the clinical medicine analysis method as described in claim 1 based on human albumin's functionalized nano silica,
It is characterized in that, in the step 4, the characterization of molecules discriminance analysis based on human albumin's nano silica are as follows:
(1) effect in ethanol/acetonitrile solution between template molecule and propylene phthalein amine molecule is analyzed by ultraviolet visible spectrometry
Power；
(2) the haemocyanin solution that various concentration is added dropwise observes ultraviolet-visible light spectral curve.
4. the clinical medicine analysis method as described in claim 1 based on human albumin's functionalized nano silica,
It is characterized in that, it is provided in an embodiment of the present invention to be based on human albumin's nano silica detection analysis in the step 5 are as follows:
(1) FITC-APTS-silica particle is added in fluorescence spectrum quartz colorimetric utensil；
(2) oneself is known that the haemocyanin solution of concentration is injected into cuvette to obtain steady-state fluorescence spectrum, carried out based on serum
The detection analysis of albumen.
5. the clinical medicine analysis method as claimed in claim 3 based on human albumin's functionalized nano silica,
It is characterized in that, the step 4 further comprises based in the characterization of molecules discriminance analysis of human albumin's nano silica:
Choose the characteristic parameter CP of several characterization of molecules interference signals based on human albumin's nano silica, including frequency F,
Time T, for observation point airspace angle Θ, polarization direction Γ and coding mode C, and using parameter as reference axis establish it is more
Coordinate system is tieed up, for each reference axis of coordinate system, respectively according to the resolution ratio of each interference characteristic parameterIt determines and corresponds to
The unit quantity of reference axis is defined as interference space based on the spatial model of establishment of coordinate system multidimensional characteristic parameter: HSI=SPACE
(CP1,CP2…CPN), any one vector in this interference spaceIt is indicated by coordinate:
Wherein CPiFor vectorThe coordinate of certain dimension in interference space is for characteristic parameter CPiSpecific value, for
The interference signal of single mode is expressed as a single vector in interference space:
For the interference signal of multimode, i.e. the one of interference signal or several characteristic parameters shows as with multivalue characteristic,
The set expression of interference characteristic vector is used in interference space:
6. the clinical medicine analysis method as claimed in claim 5 based on human albumin's functionalized nano silica,
It is characterized in that, it needs to be determined that interference signal and contrast signal are in wireless communication before several interference signal characteristic parameters of selection
Physical parameter on number field, frequency F, time T including wireless signal, for observation point airspace angle Θ, polarization direction Γ,
And coding mode C；
Characteristic parameter is used to establish space coordinates, the interference space of definition as reference axis are as follows:
HSI=SPACE (F, T, Θ, Γ, C)；
Any one vector in interference spaceIt is indicated by the coordinate in interference characteristic space:
F therein, t, θ, γ, c correspond to the coordinate value of each dimension respectively.
7. a kind of clinical medicine based on human albumin's functionalized nano silica analyzes program, operation with and terminal,
It is characterized in that, the clinical medicine analysis program based on human albumin's functionalized nano silica realizes claim 1
In the clinical medicine analysis method of human albumin's functionalized nano silica described in~6 any one.
8. a kind of terminal, which is characterized in that the terminal, which is carried, to be realized described in claim 1~6 any one based on human serum
The processor of the clinical medicine analysis method of protein function nano silica.
9. a kind of computer readable storage medium, including instruction, when run on a computer, so that computer is executed as weighed
Benefit requires the clinical medicine analysis method method described in 1-6 any one based on human albumin's functionalized nano silica.
10. the clinical medicine analysis side based on human albumin's functionalized nano silica described in a kind of implementation claim 1
The clinical medicine analysis system based on human albumin's functionalized nano silica of method, which is characterized in that it is described based on
The clinical medicine analysis system of human albumin's functionalized nano silica includes:
Nano silica prepares module, nano-silica surface processing module, the fluorescence lifetime analysis based on haemocyanin
Module, the characterization of molecules discriminance analysis module based on haemocyanin, the detection analysis module based on haemocyanin；
Nano silica preparation module is connect with nano-silica surface processing module, using TEOS as predecessor, is worked as with NH3
Make catalyst, ethyl alcohol prepares silica as solvent；
Fluorescence lifetime analysis module based on haemocyanin is connect with nano-silica surface processing module, for being based on
The fluorescence lifetime analysis of human albumin's nano silica；
Characterization of molecules discriminance analysis module based on haemocyanin is connect with nano-silica surface processing module, for carrying out
Characterization of molecules discriminance analysis based on human albumin's nano silica；
Detection analysis module based on haemocyanin is connect with nano-silica surface processing module, for carrying out based on people's blood
Albumin nano silica detection analysis.
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