CN102552944B - Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof - Google Patents

Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof Download PDF

Info

Publication number
CN102552944B
CN102552944B CN 201210016074 CN201210016074A CN102552944B CN 102552944 B CN102552944 B CN 102552944B CN 201210016074 CN201210016074 CN 201210016074 CN 201210016074 A CN201210016074 A CN 201210016074A CN 102552944 B CN102552944 B CN 102552944B
Authority
CN
China
Prior art keywords
aptes
lmp1
peg
contrast agent
nasopharyngeal carcinoma
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN 201210016074
Other languages
Chinese (zh)
Other versions
CN102552944A (en
Inventor
王维
容鹏飞
刘晟
周科朝
曾文彬
邹蟠
张声旺
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Central South University
Original Assignee
Central South University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Central South University filed Critical Central South University
Priority to CN 201210016074 priority Critical patent/CN102552944B/en
Publication of CN102552944A publication Critical patent/CN102552944A/en
Application granted granted Critical
Publication of CN102552944B publication Critical patent/CN102552944B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Landscapes

  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The invention discloses a nasopharyngeal carcinoma targeted magnetic resonance contrast agent and a preparation method thereof. The method comprises the following steps: synthesizing superparamagnetic Fe3O4 with grain diameter of between 10 and 15nm by adopting a chemical coprecipitation method, coating with APTES ((3-aminopropyl)triethoxysilane) or connecting superparamagnetic Fe3O4 to 10-15nm Fe3O4 to carry out surface amination on Fe3O4 to obtain Fe3O4-APTES surface-modified micro-particles; and connecting Fe3O4-APTES and EB (Epstein-Barr) virus latent membrane protein 1 monoclonal antibody (LMP1, Clone CS.1-4) with polyethylene glycol (PEG) as a connecting arm to obtain Fe3O4-APTES-PEG-LMP1, Clone CS.1-4 colloidal solution with stable dispersion. Compared with a conventional contrast agent, the contrast agent has the advantages of low toxicity, high stability, good biocompatibility, high sensitivity and good specific targeting property to LMP1<+>-nasopharyngeal carcinoma, and can be used for nasopharyngeal carcinoma screening and specific magnetic resonance diagnosis.

Description

A kind of nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof
Technical field
The present invention relates to the medical biotechnology Material Field, especially a kind of LMP1 +-nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof.
Background technology
Tumor is one of principal disease of nowadays influence human health, a report of World Health Organization (WHO) shows, along with the variation of global human living environment, the incidence rate of global malignant tumor will rise to for 1,500 ten thousand people/years by 1,000 ten thousand present people/years, became the No.1 killer who threatens human health.In China (particularly South China), in recent years, the sickness rate of nasopharyngeal carcinoma is in rising trend, and the nasopharyngeal carcinoma case in the whole world 80% occurs in China.
The nasopharyngeal carcinoma histological type is mainly non-keratinization type undifferentiated carcinoma, is prone to cervical lymph node and metastasis; In initial journey diagnosis and treatment patient, III-IV phase patient has accounted for 70%-80%.A very long time all is the standard care means of nasopharyngeal carcinoma to radiotherapy in the past, 5 years survival rate I-II phases for the treatment of of nasopharyngeal carcinoma are about 60%, the III-IV phase, then survival rate was very low, 20%-40% only, and treating failed main cause is local recurrence and metastasis.In addition, serious side reaction is difficult to avoid sometimes behind the Advancd Npc, therefore, and early discovery nasopharyngeal carcinoma and to carry out Accurate Diagnosis most important to its prognosis.
Epstein-Barr virus (Epstein-Barr Virus, EBV) be the Etiological of nasopharyngeal carcinoma, EBV-LMP1 (Epstein-Barr Virus Latent membrane protein 1) gene has been listed in oncogene, is unique gene that can transform people and the rodent zooblast of In vitro culture and make cell acquisition " immortality " in the EBV immutalizing gene.Wherein the latent membrane protein 1 (Latent membrane protein 1) of Epstein-Barr virus coding positive rate in nasopharyngeal cancer patient surpasses 65%, is important tumorigenesis albumen, is the present extensively significant antigen on the nasopharyngeal carcinoma cell surface of approval.Be expected to become the target of clinical diagnosis and treatment, and multiplication capacity, invasion and attack and the transfer ability of the positive nasopharyngeal carcinoma of latent membrane protein all obviously be better than the latent membrane protein Nasopharyngeal Carcinoma Patients with Negative, both also have larger difference to the reaction of oncotherapy scheme.Therefore nasopharyngeal carcinoma is carried out further typing obvious clinical meaning and scientific value.The Fe of our development 3O 4-APTES-PEG-LMP1, Clone CS.1-4 mr contrast agent not only can be used for the diagnostic imaging of nasopharyngeal carcinoma, and can utilize MRI that nasopharyngeal carcinoma is carried out further typing.
Magnetic resonance (Magnetic Resonance Imaging, MRI) has accurate soft tissue resolution because of it, the radiation damage that causes without X ray, in the monitoring of the diagnosis of tumor and curative effect, playing the part of more and more important role, mr contrast agent (Contrast agent for Magnetic resonance imaging, MRICA) can significantly improve contrast between normal and the abnormal structure, particularly improve tumor tissues and normal surrounding tissue between signal to noise ratio, improve the accuracy rate of the diagnosis of infantile tumour.Often there is larger difficulty in the Conventional MRI imaging to the early stage Accurate Diagnosis of this tumor.
At present whole world hospital uses that maximum feminine gender/positive mr contrast agent mainly is non-specific contrast medium, is divided into according to magnetization property: positive contrast medium and Negative contrast media two large classes.Positive contrast medium is mainly and contains gadolinium and contain manganic chelates, its relaxation performance is low, because free gadolinium and manganese have stronger toxicity to organism, as: the toxic reaction (can cause renal fibrosis etc.) of gadolinium (Gd) etc., and it is a kind of blood pond property contrast medium, and its scope of application is narrower.Its effect mainly is to shorten longitudinal relaxation time (T1 relaxation time), and the T1 signal of target tissue is strengthened, and is high signal in magnetic resonance, therefore, cries again the T1 agent that compares.For example: [Gd (DTPA)] .sup.2-(diethylenetriaminepentaacetate-gadolinium (III)), [Gd (DOTA)] .sup.-(1,4,7,10-tetraazacyclododecane-N, N ', N "; N ' " tetraacetate-gadolinium (III)), [Gd (BOPTA)] .sup.2-(benzylo xypropioic-diethylenetriamine pentaacetate-gadolinium (III)) and MnDPDP (N, N '-dipyridoxylethy lene diamine-N, N '-diacetate-5,5 '-bis (phosphate)-manganese (II)).Negative contrast media is named again the T2 agent that compares, and mainly is to make local magnetic field inhomogeneous, and then reduces spin relaxation time (T2 relaxation time), so that target tissue T2 phase signals lowers, the relaxation performance is high.Negative contrast media clinically is mainly take the vertical magnetic of phenanthrene as representative, but luxuriant and rich with fragrance vertical magnetic also is a kind of non-specific mr contrast agent.Compare with the gadolinium contrast medium, its characteristics are as follows: 1) can intravasation peripheral clearance, and be a kind of vascular space contrast medium, therefore, its scope of application is more extensive; 2) the per unit metallic iron can produce the change of more signal intensity, and the relaxation performance is high, especially at T 2The WI sequence; 3) have in vivo biodegradability, can be in cell homergy approach and enter ferrikinetics in the body, participate in the vital movement of ferrum in the body; 4) luxuriant and rich with fragrance vertical magnetic compass is crossed the method dyeing such as prussian blue staining, can be observed directly by light microscopic or Electronic Speculum, therefore, more likely becomes the signal component of tumor magnetic resonance target contrast agent.Shortcoming is easily to be engulfed by reticuloendothelial system and immune phagocyte in vivo, thus the half-life is short, to the active targeting aggregation capability of tumor a little less than.And to change its biocompatibility, and prolong the circulation time in its blood, strengthen its Targeting Performance, main passing through Fe 3O 4Nanoparticle is carried out surface modification.Can become face finish material have multiple, such as polysaccharide, albumin, APTES etc.Sterically hindered in order to reduce it, at Fe 3O 4And insert the linking arm of a softness between the targeted molecular as a kind of chain rope structure, its be the free-end mobility of antigen large, can be for antigen and antibodies provide relatively long time of contact and more touch opportunity in the target area, the combination between contrast agent and the target organ is just tightr.The nasopharyngeal carcinoma targeted magnetic resonance contrast agent of synthetic a kind of cheap, good biocompatibility, high specificity solves the in the past deficiency of contrast medium, has potential industrialization prospect.
Summary of the invention
The purpose of this invention is to provide a kind of LMP1 +-nasopharyngeal carcinoma specificity Negative contrast media and preparation method thereof, so that contrast medium compared with prior art has hypotoxicity, form is controlled, high stability, good biocompatibility, hypersensitivity, good specific target tropism, this contrast medium can be used for human body or non-human body LMP1 +-nasopharyngeal carcinoma strengthens contrast medium.
Purpose of the present invention can realize in the following manner
A kind of nasopharyngeal carcinoma targeted magnetic resonance contrast agent is to coat or be connected on the SPIO with 3-aminopropyl triethoxysilane (APTES), makes Fe 3O 4Surface amination obtains Fe 3O 4-APTES surface modified particulate, again with Polyethylene Glycol (PEG) as Fe 3O 4-APTES surface modified particulate and Epstein-Barr virus latent membrane protein 1 monoclonal antibody (are abbreviated as: LMP1, Clone CS.1-4; Be purchased from DAKO company, the time buying: the linking arm 2008.05.05) couples together the two the Fe of the stably dispersing that obtains 3O 4-APTES-PEG-LMP1, Clone CS.1-4 colloid solution.
Described super suitable Fe 3O 4The synthetic particle diameter of method that is the employing chemical coprecipitation is the 10-15nm granule.
Described Fe 3O 4-APTES-PEG-LMP1, Clone CS.1-4 colloidal particles particle diameter is 20-30nm.
Described nasopharyngeal carcinoma targeted magnetic resonance contrast agent is with super suitable Fe 3O 4With 3-aminopropyl triethoxysilane hybrid reaction, by centrifugal, ultrafiltration, and dialysis purification nanoparticle; Obtain Fe 3O 4Then-APTES surface modified particulate is connected with PEG; Again with LMP1, Clone CS.1-4 and with the Fe of PEG linking arm 3O 4-APTES surface modified particulate mixes, and stirs in the ice bath and spends the night; Effect by means of external magnetic field is carried out Magnetic Isolation to product, again through washing, disperses the mr contrast agent that obtains.
Fe 3O 4Reacting mol ratio with APTES is 0.1: 1-20: 1; Both reacted 30 minutes-1 hour.
Fe 3O 4The reaction mol ratio of-APTES surface modified particulate and PEG is 0.1: 1-20: 1, and both reacted 30 minutes-1 hour.
Fe with the PEG linking arm 3O 4-APTES surface modified particulate and LMP1, Clone CS.1-4 is with Fe 3O 4The mol ratio 1 of/monoclonal antibody: 1-100: 1 hybrid reaction.
Fe 3O 4Be preferably 1 with the mol ratio of APTES: 1-10: 1, Fe 3O 4Be preferably 1 with the PEG mol ratio: 1-10: 1, with the Fe of PEG linking arm 3O 4-APTES surface modified particulate and LMP1, Clone CS.1-4 is with Fe 3O 4The mol ratio of/monoclonal antibody is preferably 10: 1-100: 1.
A kind of preparation method of nasopharyngeal carcinoma targeted magnetic resonance contrast agent is with superparamagnetism Fe 3O 4With 3-aminopropyl triethoxysilane hybrid reaction, by centrifugal, ultrafiltration, and dialysis purification nanoparticle; Obtain Fe 3O 4Then-APTES surface modified particulate is connected with PEG; Again with LMP1, Clone CS.1-4 and with the Fe of PEG linking arm 3O 4-APTES surface modified particulate mixes, and stirs in the ice bath and spends the night; Effect by means of external magnetic field is carried out Magnetic Isolation to product, again through washing, disperses to obtain mr contrast agent.
Described super suitable Fe 3O 4The synthetic particle diameter of method that is the employing chemical coprecipitation is the 10-15nm granule.
Fe 3O 4Reacting mol ratio with APTES is 0.1: 1-20: 1; Both reacted 30 minutes-1 hour; Fe 3O 4The reaction mol ratio of-APTES surface modified particulate and PEG is 0.1: 1-20: 1; Both reacted 30 minutes-1 hour; Fe with the PEG linking arm 3O 4-APTES surface modified particulate and LMP1, Clone CS.1-4 is with Fe 3O 4The mol ratio 1 of/monoclonal antibody: 1-100: 1 hybrid reaction.
A kind of LMP1 +The negative mr contrast agent of-nasopharyngeal carcinoma specificity is to use the MOLECULE DESIGN platform of the computer simulation SARS drug design software buildings such as Discovery Studio and Autodock to design, and is prepared from by above method.
Advantage of the present invention:
The LMP1 of the present invention's preparation +The negative mr contrast agent of-nasopharyngeal carcinoma specificity, the Fe that produces 3O 4-LMP1 is the mr contrast agent of good biocompatibility, has superparamagnetism.Cell in vitro is learned experiment confirm, and this contrast medium has no significant effect blood cell shape, and growth has no significant effect to nephrocyte, the rat acute zoopery confirms that this contrast medium safety is good in the body, without 1 routine animal dead, can get rid of from kidney, not long-term aggregated problem in the body.Solve routine and contained the toxicity problems such as gadolinium contrast medium kidney, solved conventional Fe 3O 4Biocompatibility is poor, easily is gathered in the problem of the reticuloendothelial systems such as receptor liver spleen.
The LMP1 of the present invention's preparation +The negative mr contrast agent of-nasopharyngeal carcinoma specificity, cell in vitro is learned experiment confirm, and the magnetic cell sorting instrument shows that its sensitivity is 85.9%, and simultaneously, it has high special targeting LMP-1 +The ability of nasopharyngeal carcinoma cell (CNE1-LMP1 cell strain), specificity is 75.51%, and nude mice lotus tumor model M R imaging experiment confirms in the body, and it is to LMP-1 +The sensitivity of nasopharyngeal carcinoma tumor imaging and specificity are 85.7% and 71.4%, and have high relaxation performance, and this contrast medium mainly is being gathered in LMP-1 +Nasopharynx carcinoma kitchen range district.Solved the low relaxation performance of Conventional MRI contrast medium, to nasopharyngeal carcinoma hyposensitivity and low specificity, the problem that the MRI rate of correct diagnosis is low.
The Fe of the present invention's preparation 3O 4-APTES-PEG-LMP1, Clone CS.1-4 mr contrast agent is through APTES, PE6 and LMP1, after Clone CS.1-4 surface modification and the modification, have good dispersive property, solved particle agglomeration easily to the problem of animal dead, promote its clinical value.
Description of drawings
Fig. 1 is technical process figure of the present invention and test experience step sketch map;
Fig. 2 Fe 3O 4-APTES-PEG-LMP1, Clone CS.1-4 and LMP-1 +Nasopharyngeal carcinoma is carried out specific binding optical microscopy map (* 1000);
2-1 is that LMP1 target contrast agent and CNE1-LMP1 cell are incubated imaging under the rear oily mirror altogether;
2-2 is that LMP1 target contrast agent and CNE1 cell are incubated imaging under the rear oily mirror altogether;
Fig. 3 Fe 3O 4-APTES-PEG-LMP1, Clone CS.1-4 and LMP-1 +The nasopharyngeal carcinoma target is incubated rear MRI image altogether; Perusal can be found T 2The WI signal intensity reduces degree increasing progressively along with the cell number magnitude;
Fig. 4 Fe 3O 4-APTES-PEG-LMP1, Clone CS.1-4 is to lotus nasopharyngeal carcinoma LMP1 +With nasopharyngeal carcinoma LMP1 -Two tumor animal MRI images;
4-1,4-2 are respectively nude mice T1, T2 sequence image, and it is successful to prove that model is made tumor, and the arrow indication is the CNE+LMP1 tumor, and 4-3 is through nude mice tail vein injection LMP1-Fe 3O 4Afterwards in the T2 sequence, visible right side LMP1 +Signal obviously reduces, and left side LMP1 -Nasopharyngeal carcinoma lump signal is without significant change.
Fig. 5 lotus nasopharyngeal carcinoma LMP1 +With nasopharyngeal carcinoma LMP1 -The immunohistochemical analysis figure (* 200) of two tumor animal model LMP1 antigen presentations;
5-1 sees brown color dyeing on the LMP1+ nasopharyngeal carcinoma cell cell membrane, LMP1 expresses positive;
5-2 is that the LMP1-nasopharyngeal carcinoma cell is expressed the LMP1 feminine gender;
Fig. 6 Fe304-APTES-PEG-LMP1 is after the Clone CS.1-4 enhanced ct scans, to lotus nasopharyngeal carcinoma LMP1 +With nasopharyngeal carcinoma LMP1 -Two tumor animal Prussian blue colored graphs;
6-1 sees in the CNE1-LMP1 transplanted tumor lump tissue that indigo plant a large amount of, clear display dyes the ferrum granule, illustrate that the LMP1 target contrast agent is assembled well in the tumor tissues; Only visible a little indigo plant is dyed ferrum granule (shown in the arrow) in the 6-2, CNE1 transplanted tumor lump tissue.Compare obvious difference with front figure, thereby show LMP1 targeting Fe 3O 4Has certain targeting effect for nude mouse lotus human nasopharyngeal carcinoma CNE1-LMP1 transplanted tumor lump.
The specific embodiment
Be intended to further specify the present invention below in conjunction with embodiment, and unrestricted the present invention.
Embodiment 1
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 0.1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 0.1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 1: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (oleic acid acyl, consumption 0.2%~0.5%) ultra-sonic dispersion after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 2
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 0.1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 0.1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 10: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the oleic acid acyl. consumption 0.2%~0.5%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 3
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 0.1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 0.1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 1: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the hexenyl bis-stearamides. consumption is 0.5%~2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.。
Embodiment 4
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 1: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the hexenyl bis-stearamides. consumption is 0.5%~2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.。
Embodiment 5
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 10: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (PVAC polyvinylalcohol. consumption is 2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 6
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 1: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 1: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 100: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (PVAC polyvinylalcohol. consumption is 2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 7
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 10: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 10: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 1: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (oleic acid. consumption 0.2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 8
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 10: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 10: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 10: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the oleic acid acyl. consumption 0.2%~0.5%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 9
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 10: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 10: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 100: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the oleic acid acyl. consumption 0.2%~0.5%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 10
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 20: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 20: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 1: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (the oleic acid acyl. consumption 0.2%~0.5%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 11
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 20: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 20: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 10: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (oleic acid. consumption 0.2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.
Embodiment 12
Getting 0.5mmoL ferrous chloride and 1.2mmoL ferric chloride in aqueous solution mixes, add 15mL dodecylbenzene sodium sulfonate (DBS) solution (25mM) for preparing in advance, after solution becomes milky, add 75mL toluene, high-speed stirred 30min allows the solution mix homogeneously, drip concentration and be the NaOH solution adjust pH of 2.5M to 8-9, continue to stir 1h.Change mixed solution over to the separatory funnel standing demix, visible particle is scattered in the upper toluene solution, divide to fall water layer, the upper strata black liquor is changed in the there-necked flask, this moment is logical argon deoxygenation first, then slowly heats up, and refluxes half an hour after steaming water in the solution and part toluene, again with the 3-aminopropyl triethoxysilane according to mol ratio 20: 1, be dissolved in the water and mix.This mixture is placed the pure argon environment, reflux half an hour, make the magnetic iron ore crystallization, by centrifugal, ultrafiltration, and dialysis purification nanoparticle.Produce evengranular nanoparticle, then react half an hour at 20: 1 according to mol ratio with the PEG linking arm, carry out covalent coupling, again with Fe 3O 4The mol ratio of/monoclonal antibody is got the Fe of the APTES modification of PEG connection at 100: 1 3O 4Nanometer magnetofluid and LMP1, Clone CS.1-4 mixes, and places ice bath, and stirring is spent the night.Effect by means of external magnetic field is carried out Magnetic Isolation to product, alternately washs 4-5 time with ultra-pure water and dehydrated alcohol, is scattered in after impurity is removed in dialysis in the ultra-pure water, just obtains the Fe of nasopharyngeal carcinoma antibody modification 3O 4Nanometer magnetofluid, particle diameter 20-30nm; Add dispersant (oleic acid. consumption 0.2%) ultra-sonic dispersion is after 30 minutes, the concentration that obtains described mr contrast agent is 0.25mmol/ml.

Claims (2)

1. a nasopharyngeal carcinoma targeted magnetic resonance contrast agent is characterized in that, is to coat or be connected on the SPIO with the 3-aminopropyl triethoxysilane, makes Fe 3O 4Surface amination obtains Fe 3O 4-APTES surface modified particulate, again with Polyethylene Glycol as Fe 3O 4Linking arm between-APTES surface modified particulate and Epstein-Barr virus latent membrane protein 1 monoclonal antibody couples together the two the Fe of the stably dispersing that obtains 3O 4-APTES-PEG-LMP1, Clone CS.1-4 colloid solution;
Described super suitable Fe 3O 4The synthetic particle diameter of method that is the employing chemical coprecipitation is the 10-15nm granule;
Fe 3O 4-APTES-PEG-LMP1, Clone CS.1-4 colloidal particles particle diameter is 20-30nm;
Described nasopharyngeal carcinoma targeted magnetic resonance contrast agent is with superparamagnetism Fe 3O 4With 3-aminopropyl triethoxysilane hybrid reaction, by centrifugal, ultrafiltration, and dialysis purification nanoparticle; Obtain Fe 3O 4Then-APTES surface modified particulate is connected with PEG; Again with LMP1, Clone CS.1-4 and with the Fe of PEG linking arm 3O 4-APTES surface modified particulate mixes, and stirs in the ice bath and spends the night; Effect by means of external magnetic field is carried out Magnetic Isolation to product, again through washing, disperses the mr contrast agent that obtains;
Fe 3O 4Reacting mol ratio with APTES is 0.1:1-20:1; Both reacted 30 minutes-1 hour;
Fe 3O 4The reaction mol ratio of-APTES surface modified particulate and PEG is 0.1:1-20:1; Both reacted 30 minutes-1 hour;
Fe with the PEG linking arm 3O 4-APTES surface modified particulate and LMP1, Clone CS.1-4 is with Fe 3O 4The mol ratio 1:1-100:1 hybrid reaction of/monoclonal antibody.
2. the preparation method of a nasopharyngeal carcinoma targeted magnetic resonance contrast agent as claimed in claim 1 is characterized in that, with superparamagnetism Fe 3O 4With 3-aminopropyl triethoxysilane hybrid reaction, by centrifugal, ultrafiltration, and dialysis purification nanoparticle; Obtain Fe 3O 4Then-APTES surface modified particulate is connected with PEG; Again with LMP1, Clone CS.1-4 and with the Fe of PEG linking arm 3O 4-APTES surface modified particulate mixes, and stirs in the ice bath and spends the night; Effect by means of external magnetic field is carried out Magnetic Isolation to product, again through washing, disperses to obtain mr contrast agent;
Described super suitable Fe 3O 4The synthetic particle diameter of method that is the employing chemical coprecipitation is the 10-15nm granule;
Fe 3O 4Reacting mol ratio with APTES is 0.1:1-20:1; Both reacted 30 minutes-1 hour; Fe 3O 4The reaction mol ratio of-APTES surface modified particulate and PEG is 0.1:1-20:1; Both reacted 30 minutes-1 hour; Fe with the PEG linking arm 3O 4-APTES surface modified particulate and LMP1, Clone CS.1-4 is with Fe 3O 4The mol ratio 1:1-100:1 hybrid reaction of/monoclonal antibody.
CN 201210016074 2012-01-18 2012-01-18 Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof Active CN102552944B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 201210016074 CN102552944B (en) 2012-01-18 2012-01-18 Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 201210016074 CN102552944B (en) 2012-01-18 2012-01-18 Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof

Publications (2)

Publication Number Publication Date
CN102552944A CN102552944A (en) 2012-07-11
CN102552944B true CN102552944B (en) 2013-03-27

Family

ID=46400392

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 201210016074 Active CN102552944B (en) 2012-01-18 2012-01-18 Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof

Country Status (1)

Country Link
CN (1) CN102552944B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103290019B (en) * 2013-06-14 2014-03-12 严鹏科 Atherosclerosis targeting aptamer as well as preparation method and application thereof
CN104090050B (en) * 2014-07-08 2015-10-28 复旦大学 A kind of memebrane protein covalency fixing means based on magnetic material

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101712710A (en) * 2008-10-08 2010-05-26 中南大学 LMP1targeted oligonucleotide and application thereof serving as radiotherapeutic sensitizer
CN102174516A (en) * 2011-01-20 2011-09-07 中南大学 Molecular target for diagnosing and treating nasopharyngeal cancer related with Epstein-Barr virus (EBV) infection and application thereof
CN102249345B (en) * 2011-04-02 2013-05-29 浙江大学 Preparation method of Exendin-4 coupling superparamagnetic iron oxide nano particle

Also Published As

Publication number Publication date
CN102552944A (en) 2012-07-11

Similar Documents

Publication Publication Date Title
Bulte Superparamagnetic iron oxides as MPI tracers: A primer and review of early applications
Ahrens et al. Tracking immune cells in vivo using magnetic resonance imaging
Zhang et al. Noninvasive monitoring of orthotopic glioblastoma therapy response using RGD-conjugated iron oxide nanoparticles
Pinho et al. Relaxometric studies of γ-Fe2O3@ SiO2 core shell nanoparticles: when the coating matters
Bai et al. Synthesis of ultrasmall Fe3O4 nanoparticles as T 1–T 2 dual-modal magnetic resonance imaging contrast agents in rabbit hepatic tumors
CN101912623B (en) Preparation and application of Fe-Gd double-mode magnetic resonance contrast agent with targeting function
Zhao et al. A GPC3-specific aptamer-mediated magnetic resonance probe for hepatocellular carcinoma
CN103143041B (en) Preparation method of targeted MRI (magnetic resonance imaging) contrast medium based on folic acid modified iron oxide nanoparticles
Shi et al. Hemoglobin-mediated biomimetic synthesis of paramagnetic O2-evolving theranostic nanoprobes for MR imaging-guided enhanced photodynamic therapy of tumor
CN105920620A (en) Magnetic fluorescent multimodal nano biological probe as well as preparation method and application thereof
Wu et al. Reduction-active Fe3O4-loaded micelles with aggregation-enhanced MRI contrast for differential diagnosis of Neroglioma
Chen et al. Improving sensitivity of magnetic resonance imaging by using a dual-targeted magnetic iron oxide nanoprobe
Khmara et al. Preparation of poly-L-lysine functionalized magnetic nanoparticles and their influence on viability of cancer cells
JP2015511938A (en) Superparamagnetic nanoparticles as contrast agent (T2 *) in magnetic resonance imaging (MRI) of magnetic sensitivity (MAGNETICSUSCEPTIBILITY)
CN104548142A (en) Preparation method of hyaluronic acid modified superparamagnetic iron oxide/gold composite nanoprobe
CN105079826A (en) Preparation method and application of RGD@BBN double-targeted MR (magnetic resonance)/optical dual-mode molecular probe
Luo et al. Hyaluronic acid-mediated multifunctional iron oxide-based MRI nanoprobes for dynamic monitoring of pancreatic cancer
Baroni et al. In vivo assessment of tumour associated macrophages in murine melanoma obtained by low-field relaxometry in the presence of iron oxide particles
CN106466488A (en) There is ultra-fine magnetic core-shell nano material and its preparation and the application of tumor cell targeting
CN102552944B (en) Nasopharyngeal carcinoma targeted magnetic resonance contrast agent and preparation method thereof
Li et al. Designing Smart Iron Oxide Nanoparticles for MR Imaging of Tumors
CN102350002B (en) Glioma-targeting molecule magnetic resonance contrast agent as well as preparation method and application thereof
CN101474414B (en) Preparation and application of polymer-coated magnetic nanoparticle contrast agent
Jiang et al. Noninvasively characterizing the different αvβ3 expression patterns in lung cancers with RGD-USPIO using a clinical 3.0 T MR scanner
Legacz et al. Contrast agents and cell labeling strategies for in vivo imaging

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant