CN104560009A - Preparation method of herceptin modified and quantum dot/ferroferric oxide loaded magnetic fluorescent silica nanoparticles - Google Patents
Preparation method of herceptin modified and quantum dot/ferroferric oxide loaded magnetic fluorescent silica nanoparticles Download PDFInfo
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- CN104560009A CN104560009A CN201310485311.7A CN201310485311A CN104560009A CN 104560009 A CN104560009 A CN 104560009A CN 201310485311 A CN201310485311 A CN 201310485311A CN 104560009 A CN104560009 A CN 104560009A
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Abstract
The invention relates to a preparation method of herceptin modified and quantum dot/ferroferric oxide loaded magnetic fluorescent silica nanoparticles. The prepared composite nanoparticles can be used for separating and detecting circulating tumor cells. The preparation method of the herceptin modified and quantum dot/ferroferric oxide loaded magnetic fluorescent silica nanoparticles comprises steps as follows: a POSS (polyhedral oligomeric silsesquiaxana) conjugated quantum dot is prepared firstly, the POSS conjugated quantum dot is coated with silica spheres, Br-IO is conjugated to the outer layer of QD-SiNP, finally, herceptin is conjugated to IO-QD-SiNP, and the herceptin modified and quantum dot/ferroferric oxide loaded magnetic fluorescent silica nanoparticles are obtained. The composite nanoparticles prepared with the method have fluorescence, magnetism and target functions and are expected to be applied to in-vivo and in-vitro bioimaging.
Description
Technical field
The present invention relates to the preparation method of the quantum dot-silicon-dioxide/ferriferrous oxide nano grain of the polyhedral oligomeric silsesquioxane conjugation that a kind of Trastuzumab is modified.
Background technology
Cancer is the principal element of world population death, and the characteristic being easy to shift due to it causes it to be difficult to cure, and this means that cancer cells can be separated from initial tumour cell and arrives blood vessel, and then arrive the different tissues of human body.Cancer cells is separated from initial tumor tissues, and then in blood vessel around, circulation is exactly the circulating tumor cell that we know.The early diagnosis that these cells of separation and detection will help us to carry out cancer, the monitoring of this diagnostic result for patient and result for the treatment of is all vital.
Have several method at present for separation and the counting of circulating tumor cell, such as immunomagnetic beads cell sorting and fluorescence-activated cell sorting are two kinds of the most frequently used business methods.But these methods meet with the return rate of very low tumour cell, and some in the middle of them need to carry out cell dyeing work complicated further with fluorescence antibody before testing.Needs are spent the more cell cultures time by such a treating processes, also can reduce the precision of detection simultaneously.The micro-fluidic technologies that can be used for cellular segregation at present shows fine prospect.Compared with being separated, longer duration of contact and stronger interaction between the antibody modified by cell and microfluidic surface, can obtain higher restoring degree in batches.But this technology is needed complicated building-up process and modified especially by antibody, the cell of simultaneously catching needs to be separated for detection usually.Recently, Viovy seminar reports a kind of new microfluid, and it has tied the advantage of microfluid and magnetic field cellular segregation.This is a series of biological functional ultra paramagnetic particles, the self-assembly carried out in microchannel with magnetic field model.The magnetic glass powder of this new application of installation replaces some polymkeric substance, and this more easily synthesizes and uses antibody modification, also therefore show extraordinary industrial prospect.
Use nanotechnology as hinge by fluorescence, magnetic and selectively targeted three kinds of performances combine screening and the detection that better can realize cancer cells.Owing to having Z 250 in nanoparticle, these nanoparticles are separated from blood easily via magnetic field, therefore may be used for cellular segregation; Due to the fluorescence of quantum dot in nanoparticle, screening process can dynamically be observed, and can directly realize whole observation process; Because nanoparticle surface has specific antibody to modify, by the initiative recognition effect of antibody, the cancer cells of particular types is easier to be separated from various kinds of cell.The simplification of cell detection, the selectively targeted of visual and cancer cells of cell screening will improve the survival rate of cell and the precision of detection.Therefore this collection fluorescence, magnetic and target function are applicable in body and external biological imaging applications in the Multifunctional composite nanometer grain of one very much.
Summary of the invention
The object of the invention is to the preparation method of the quantum dot-silicon-dioxide/ferriferrous oxide nano grain proposing the polyhedral oligomeric silsesquioxane conjugation that a kind of Trastuzumab of preparation is modified, preparation method provided by the invention is: the quantum dot first preparing polyhedral oligomeric silsesquioxane (POSS) conjugation, make it have higher quantum yield, use the quantum dot of the coated POSS conjugation of mesoporous silica spheres subsequently, for quantum dot provides one to stablize and firm structure, also can provide an activating surface for further finishing simultaneously, then by the skin of Br-IO conjugation to QD-SiNP, silica dioxide nano particle is made to have magnetic, finally by the silica dioxide nano particle of iron oxide layer Trastuzumab functionalization, using Trastuzumab as target part, can in order to the cancer cells of active targeting HER2 overexpression.The composite nano-granule synthesized has fluorescence, magnetic and target function, has very high optical strength, stable photostabilization and relatively low toxicity, is applicable in body and external biological imaging applications.
The technical scheme that the present invention solves the quantum dot-silicon-dioxide/ferriferrous oxide nano grain technical problem of the polyhedral oligomeric silsesquioxane conjugation that a kind of Trastuzumab of described preparation is modified is, design one can by ferriferrous oxide nano grain conjugation to the outer field method of mesoporous silicon oxide coated quantum dots nanoparticle, concrete technology is: modified by the quantum dot POSS synthesized, join in the sodium hydroxide mixing solutions of 75ml water and 4ml2M subsequently, be heated to 75 DEG C, then 3ml tetraethoxy and 3ml ethyl acetate are joined in above-mentioned reaction soln and stir 3 hours, above-mentioned for 10mg gained silica dioxide nano particle and 4ml triethoxyl silane are dissolved in 10ml dehydrated alcohol, back flow reaction at 70 DEG C, finally by 1000N Fe
3o
4be dissolved in the ethanolic soln containing 20 μ L triethylamines with the silica dioxide nano particle being loaded with quantum dot, at 75 DEG C, nitrogen protection reacts 24 hours, by reacted product ethanol centrifuge washing, the quantum dot-silicon-dioxide/ferriferrous oxide nano grain of polyhedral oligomeric silsesquioxane conjugation can be obtained.
Present invention employs a kind of multifunctional nano system that can be used for cellular segregation and detection, it combines the advantage of the cellular segregation of immune magnetic and active fluoro, both can be used for direct batch treatment, can be used for again microfluid newly developed.This composite nano-granule has very high optical strength, stable photostabilization and relatively low toxicity, is therefore applicable to very much in body and external biological imaging applications.
Embodiment
The present invention is described further below in conjunction with embodiment.Specific embodiment does not limit claim of the present invention.
Quantum dot-silicon-dioxide/ferriferrous oxide nano grain preparation method the technical scheme of the polyhedral oligomeric silsesquioxane conjugation that a kind of Trastuzumab of preparation of the present invention is modified is, multistep processes is adopted to prepare composite nano-granule, concrete technology is: the quantum dot first preparing POSS conjugation by oil-in-water method, the quantum dot nano ball of the coated POSS conjugation of silica spheres is prepared subsequently by tetraethoxy and ethyl acetate, then by the Br-IO conjugation prepared with the bromo acid skin to QD-SiNP, finally by Trastuzumab conjugation on IO-QD-SiNP, quantum dot-silicon-dioxide/Z 250 the composite nano-granule of the polyhedral oligomeric silsesquioxane conjugation that Trastuzumab is modified can be obtained.
Quantum dot-silicon-dioxide/Z 250 the composite nano-granule of the polyhedral oligomeric silsesquioxane conjugation that the Trastuzumab obtained of the present invention is modified.In the scope of applicant's retrieval, there are no the bibliographical information of this magnetic fluorescence target composite nano-granule and preparation method thereof.Quantum dot-silicon-dioxide/Z 250 the composite nano-granule of the polyhedral oligomeric silsesquioxane conjugation of modifying due to Trastuzumab of the present invention again has fluorescence concurrently, magnetic and target function, is expected to the separation and detection for circulating tumor cell.
The present invention does not address part and is applicable to prior art.
Embodiment 1:
Be dissolved in chloroform by the quantum dot of 50 μ L, its concentration is 50 μMs, pours in above-mentioned solution subsequently by the 0.55mM CTAB aqueous solution of 5ml, then by gained solution vigorous stirring 30 minutes, forms oil-in-water microemulsion.Subsequently the aqueous solution is heated to 60 DEG C, to evaporate removing chloroform, then 3ml tetraethoxy and 3ml ethyl acetate is joined in above-mentioned reaction soln and stir 3 hours.The silica dioxide nano particle being loaded with quantum dot with ethanol repeated washing, to remove the complete reactant of unreacted, can obtain the nanometer ball of silicon dioxide-coated quantum dots.Embodiment 2:
Finishing is carried out to silicon-dioxide, 10mg silica dioxide nano particle and 4ml triethoxyl silane is dissolved in 10ml dehydrated alcohol, back flow reaction 3 hours at 70 DEG C.Finishing is carried out to Z 250, by 1.3g BMPA and 15mg Fe
3o
4nanoparticle is dissolved in 10ml normal hexane, reacts and flows down lasting 72 hours in nitrogen protection.In order to by ferriferrous oxide nano grain conjugation on the silica dioxide nano particle being loaded with quantum dot, by 1000N Fe
3o
4be dissolved in the ethanolic soln having 20 μ L triethylamines with the silica dioxide nano particle being loaded with quantum dot, reactant is heated to 75 DEG C of back flow reaction 24 hours under nitrogen protection, the reactant ethanol centrifuge washing obtained removes unnecessary Fe
3o
4, the nanoparticle of the silicon dioxide-coated quantum dots of Z 250 conjugation can be obtained.
Embodiment 3:
Trastuzumab is dissolved in PBS solution and is diluted to 1mg/ml, the at room temperature appropriateness concussion of antibody and nanoparticle to be spent the night mixing, after further washing by centrifugal and PBS solution, the quantum dot-silicon-dioxide/Z 250 composite nano-granule of the polyhedral oligomeric silsesquioxane conjugation that Trastuzumab is modified can be obtained.
Claims (4)
1. the preparation method of the polyhedral oligomeric silsesquioxane conjugation quantum dot-silicon-dioxide/ferriferrous oxide nano grain of a Trastuzumab modification, it is characterized in that method provided by the invention is: the quantum dot first preparing polyhedral oligomeric silsesquioxane (POSS) conjugation, use the quantum dot (QD-SiNP) of the coated POSS conjugation of silica spheres subsequently, then by Z 250 (Br-IO) conjugation of modifying with the bromine skin (IO-QD-SiNP) to QD-SiNP, finally by Trastuzumab conjugation on IO-QD-SiNP, finally obtain polyhedral oligomeric silsesquioxane conjugation quantum dot-silicon-dioxide/ferriferrous oxide nano grain that Trastuzumab is modified, a kind of target composite nano-granule with fluorescence and magnetic.
2. the preparation method of the polyhedral oligomeric silsesquioxane conjugation quantum dot-silicon-dioxide/ferriferrous oxide nano grain of a kind of Trastuzumab modification according to claim 1, is characterized in that described quantum dot is that polyhedral oligomeric silsesquioxane is modified.
3. the preparation method of the polyhedral oligomeric silsesquioxane conjugation quantum dot-silicon-dioxide/ferriferrous oxide nano grain of a kind of Trastuzumab modification according to claim 1, is characterized in that described Z 250 is that Br modifies.
4. the preparation method of the polyhedral oligomeric silsesquioxane conjugation quantum dot-silicon-dioxide/ferriferrous oxide nano grain of a kind of Trastuzumab modification according to claim 1, is characterized in that described nanoparticle is that Trastuzumab is modified.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106421813A (en) * | 2016-10-21 | 2017-02-22 | 新乡医学院 | Drug-loaded nanoparticle with double targeting function and preparation method and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106421813A (en) * | 2016-10-21 | 2017-02-22 | 新乡医学院 | Drug-loaded nanoparticle with double targeting function and preparation method and application thereof |
| CN106421813B (en) * | 2016-10-21 | 2019-07-26 | 新乡医学院 | Medicine-carried nano particles and its preparation method and application with dual-target function |
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Application publication date: 20150429 |