CN102636649A - Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material - Google Patents
Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material Download PDFInfo
- Publication number
- CN102636649A CN102636649A CN2012100626675A CN201210062667A CN102636649A CN 102636649 A CN102636649 A CN 102636649A CN 2012100626675 A CN2012100626675 A CN 2012100626675A CN 201210062667 A CN201210062667 A CN 201210062667A CN 102636649 A CN102636649 A CN 102636649A
- Authority
- CN
- China
- Prior art keywords
- cea
- antibody
- nano material
- antigen
- conversion fluorescence
- 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.)
- Granted
Links
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention relates to a kit for detecting a carcinoembryonic antigen (CEA) based on an antibody functionalized magnetic nanometer material and an up-conversion fluorescence nanometer material. The kit provided by the invention is characterized in that an anti-CEA first antibody (capturing antibody) functionalized magnetic nanometer material is used to enrich and separate CEAs in a sample; moreover, the up-conversion fluorescence nanometer material NaY0.78F4:Yb0.20 is marked with an anti-CEA second antibody (detecting antibody) based on the characteristics of the high laser-induced fluorescence sensitivity of the up-conversion fluorescence nanometer material and the capability of effectively avoiding biological background fluorescence interference of the sample, and Ho0.02 is used as a detecting probe; and the high-sensitively, stable and rapid CEA detecting method is established by combining the two nanometer materials and an antigen-antibody specificity reaction principle. The kit can be used for detecting the content of CEA in clinic or in scientific researches and is used for auxiliary tumor diagnosis, guide therapy and prompt prognosis.
Description
Technical field
The invention belongs to nano material and biological technical field, for a kind of based on antibody function magnetic Nano material and up-conversion fluorescence nano material and kit that conjugated antigen-antibody specificity reaction pair carcinomebryonic antigen (CEA) detects.Characteristics are to resist-and the magnetic Nano material of CEA first antibody (capture antibody) functionalization carries out separation and concentration to antigen in the sample, makes detection sensitivity improve; The up-conversion fluorescence nano material NaY that utilizes anti--CEA SA (detection antibody) to modify simultaneously
0.78F
4: Yb
0.20, Ho
0.02The thing that serves as a mark induces the up-conversion fluorescence signal further to improve the sensitivity of detection through detection laser.
Background technology
Antigen, being meant to stimulate body to produce specific immune response, and can the material of specific reaction take place with the immune response product---antibody and sensitized lymphocyte combine in vivo and in vitro.Tumor associated antigen refers in kinds of tumor cells to express to be increased, and in normal cell the synthetic antigen molecule of trace only.Carcinomebryonic antigen (CEA) is the most frequently used tumor associated antigen, and it is a kind of acidoglycoprotein, and synthesize at small intestine, liver, pancreas embryonic period, embryonic phase, and only trace exists in normal human serum.CEA is considered to the mark (60%-90% patient's rising) of colorectal cancer the earliest, but finds also to have among cancer of pancreas (～80%), cancer of the stomach (～60%), lung cancer (～75%) and breast cancer (～60%) patients serum higher CEA to express later on.CEA estimates according to all having important value curative effect judgement, PD, monitoring and the prognosis of tumor in digestive tract and lung cancer as a broad spectrum activity tumor markers.Change of serum C EA detects has become the most widely used tumour auxiliary diagnosis and prognosis judging means at present.
Utilizing Ag-Ab specific reaction principle exploitation antigen, detection of antibodies method is the basic ideas of present immunology detection, also is the basis that comprises the various tumor associated antigens detections of CEA.Traditional CEA detection method mainly contains methods such as ELISA (ELISA), electrochemiluminescence immunoassay, radiommunoassay.These methods all exist not enough separately at aspects such as optical property, stability, susceptibility, security, detection time or instrument requirement.All not good enough like elisa assay sensitivity, stability and repeatability; Although the electrochemiluminescence immunoassay has improved detection sensitivity, exist functional electrode to prepare deficiencies such as difficulty and running time length; Radiommunoassay is then because its large-scale promotion that needed radiation proof drawbacks limit.So be necessary to develop a kind of quick and convenient, good stability, highly sensitive, high specificity, the novel detection method that use cost is low.
Relation between nano material and the immunological technique is more and more tightr in recent years, and the development prospect of novel nano-material in biomedicine detects is also more and more wide.Wherein upload and change fluorescent nano material and its unique characteristic of magnetic Nano material enjoys people to pay close attention to.In rear-earth-doped inorganic nano material, one type of special luminescent material is arranged, it can convert long-wave radiation (near infrared light) to shortwave radiation through multi-photon mechanism, launches the fluorescence (ultraviolet-visible) than excitation light wave length, i.e. up-conversion fluorescence.Up-conversion has following plurality of advantages: photochemical stable; Detection background is low, and penetrability is strong, and can regulate the different up-conversion fluorescence of nano particle emission through selecting substrates with different material and dopant ion; The single excitation multi of real realization is penetrated, thereby helps detecting simultaneously polycomponent.In view of above-mentioned advantage, up-conversion nano material has become a kind of outstanding biological labled material at present.Simultaneously, in the detection of some biological sample, because the measured target content of material is very low, mark is complicated with the separating step in detecting, and the separation of mark sample and bioaccumulation efficiency will directly influence the sensitivity and the accuracy of detection.Fe
3O
4Be to use magnetic material the most widely, the Fe that in liquid, exists with the magnetic fluid form
3O
4Directed moving can take place in magnetic-particle under the effect in external magnetic field.And Fe
3O
4The stable performance of magnetic magnetic nano-particle is prone to preparation, can be connected with various biomolecules through finishing, can under the effect of externally-applied magnetic field, realize that the immobilization of these molecules or enrichment concentrate.Therefore, utilize magnetic Nano material can fast and effeciently separate and the enrichment target substance, have the specificity height, separate fast, high repeatability and other advantages with up-conversion nano material.
The at first synthetic Fe that has obtained the surface biological functionalization of the present invention
3O
4Magnetic Nano material and NaY
0.78F
4: Yb
0.20, Ho
0.02The up-conversion fluorescence nano material; Anti--CEA the first antibody (capture antibody) that combines through magnetic Nano material again utilizes the antigen-antibody reaction specificity to catch the CEA in the sample to be tested; And separate in addition concentration and separation through magnetic, combines to form the nano-particle complex of CEA capture antibody/CEA/CEA detection antibody sandwich structure appearance again with the up-conversion fluorescence nano particle of anti--CEA SA (detection antibody) mark.Inducing up-conversion fluorescence with 980nm laser excitation is detection signal, with the typical curve of setting up of variable concentrations CEA standard items.Under the 980nm induced with laser, NaY
0.78F
4: Yb
0.20, Ho
0.02Luminous signal (wavelength is 542nm) that the up-conversion fluorescence nano material produces and the amount positive correlation of CEA are to reach containing the purpose that the CEA sample carries out detection by quantitative.This invention can be used for CEA Determination on content in the human serum sample.
Summary of the invention
The present invention is a kind of kit based on antibody function magnetic Nano material magnetic separation-up-conversion fluorescence nano material marker detection CEA, and said kit comprises:
Magnetic Nano material as anti--CEA first antibody (capture antibody) functionalization of antigen capture and magnetic separation agent; Finishing the fluorescence probe that constitutes of the up-conversion fluorescence nano material of anti--CEA SA (detection antibody); Positive control; Negative control; The antigen standard items.
The CEA standard items are the CEA antigen freeze-dried powder of known quality, and positive control is for containing the human serum of finite concentration (1-10ng/mL) CEA, and negative control is PBS damping fluid (pH=7.4).
Prepare NaY respectively
0.78F
4: Yb
0.20, Ho
0.02Up-conversion fluorescence nano material and amidized Fe
3O
4Magnetic nano-balls carries out surface-functionalized modification to nano particle, subsequently with amidized Fe
3O
4Magnetic nano-balls combines with anti--CEA first antibody, and be used as and catch and the magnetic separation agent, and with NaY
0.78F
4: Yb
0.20, Ho
0.02The up-conversion fluorescence nano material combines with anti--CEA SA (detection antibody), constitutes the detector probe of mark.When in CEA capture antibody functional magnetic nano material system, adding CEA; Both combine through antigen-antibody reaction generation specificity; Through washing and magnetic separating step and after removing supernatant; The up-conversion fluorescence nano material that adds CEA detection antibody labeling; The fluorescence probe that the magnetic Nano material that CEA detects antibody labeling detects antibody labeling through antigen-antibody reaction and CEA combines to form the nano-particle complex of CEA capture antibody/CEA/CEA detection antibody sandwich structure appearance, through washing with after magnetic separates, again with 980nm induced with laser NaY
0.78F
4: Yb
0.20, Ho
0.02The up-conversion fluorescence nano material is luminous.Luminous signal intensity is relevant with CEA content within the specific limits, based on this CEA standard items is detected, and sets up typical curve, to reach containing the purpose that the CEA sample carries out detection by quantitative.
Beneficial effect:
(1) the present invention utilizes 980nm induced with laser up-conversion fluorescence to be detection signal; No bias light and autofluorescence disturb; The immune analysis technique background light that has solved FITC, RB200 labelled antibody disturbs and selectivity not good shortcoming; Lower signal to noise ratio (S/N ratio) greatly, improved detection stability and specificity.
(2) the present invention's magnetic nano-balls of utilizing the CEA antibody functionization carries out separation and concentration to target antigen in the sample and concentrates, and detection sensitivity is high, sense cycle is short, and simple to operate.
Description of drawings
Fig. 1: based on the experimental principle figure of antibody function magnetic Nano material magnetic separation-up-conversion fluorescence nano material marker detection CEA.
Fig. 2: (a): NaY
0.78F
4: Yb
0.20, Ho
0.02Up-conversion fluorescence nano material Electronic Speculum figure; (b) NaY
0.78F
4: Yb
0.20, Ho
0.02, SiO
2Up-conversion fluorescence nano material Electronic Speculum figure.
Fig. 3: (a): the ultra-violet absorption spectrum of anti--CEA one anti-(0.1mg/mL) and modification back magnetic Nano material thereof, wherein A is anti-for anti--CEA one, and B is anti--CEA one anti-back magnetic Nano material of modifying; (b): the ultra-violet absorption spectrum of anti--CEA two anti-(0.15mg/mL) and modification back up-conversion fluorescence nano material thereof, wherein A is anti-for anti--CEA two, and B is anti--CEA two anti-back up-conversion fluorescence nano materials of modifying.
Fig. 4: antibody and ultra-violet absorption spectrum after nano particle combines.A: anti--CEA one anti-and magnetic nanoparticle (2mgmL
-1) combine b: anti--CEA two anti-and up-conversion fluorescence nano material (2mgmL
-1) combine.
Fig. 5: CEA examination criteria curve map, concentration range is 2.5 * 10
-12-1 * 10
-8GmL
-1
Embodiment
Embodiment 1: the preparation of the magnetic Nano material of finishing and last conversion nano particle
(1) utilization hydro-thermal-solvent thermal technology preparation NaY
0.78F
4: Yb
0.20, Ho
0.02Last conversion nano particle, and it is done finishing.Take by weighing 0.18g Y
2O
3(28%), 0.788g Yb
2O
3(70%) and 0.022g Er
2O
3(2%) potpourri adds heating for dissolving in the nitric acid; Obtain the nitrate of rare earth element powder after vaporing away redundant nitric acid; It is dissolved in the 8mL deionized water; Add 1.0635g two ethylenediamine hydrate Sequestrene AAs (EDTA) again, and regulate pH, form the EDTA-Ln solution of clear to alkalescent.Get 25mL monoethylene glycol, add 3mL HF and 0.4g cetyl trimethyl ammonium bromide (CTAB), add the above-mentioned EDTA-Ln solution of 8mL while stirring, obtain white liliquoid.Add the 5.5mL red fuming nitric acid (RFNA) at last, after stirring, transfer in the 50mL band teflon-lined agitated reactor 195 ℃ of reaction 24h.After reaction finishes, be allowed to condition at and naturally cool to room temperature in the air, abandon supernatant liquid, the solid at the bottom of the still with the hot water injection in beaker, ultrasonic 10 minutes, left standstill then several minutes, discard supernatant liquid, it is ultrasonic to add hot water again, repeats 3 times.Add the ultrasonic dispersion of ethanol then, last centrifugal gained solid places 70 ℃ of oven drying 10h, and pressed powder stores for future use.
(2) utilizing improved
method that up-conversion nanoparticles is carried out surface amination modifies.
Up-conversion fluorescence nano material adding after the 60mg processing is filled in the 250mL conical flask of 60mL isopropyl alcohol, and ultrasonic 40min reaches fully and disperses.Under quick magnetic agitation, add the ammoniacal liquor of 20mL distilled water, 2.5mL 25% successively, seal.Under 35 ℃, add the mixed solution that dropwise adds 20mL isopropyl alcohol and 50 μ L ethyl orthosilicates behind the magnetic agitation 10min.Behind the reaction 3h, dropwise add the mixed solution of the 3-aminopropyl triethoxysilane of 30mL isopropyl alcohol and 200 μ L again, continue to stop to stir ageing 2h under the room temperature behind the reaction 1h.With the product centrifuging, washing is repeatedly dried 12h for 60 ℃ at last, and obtaining the surface has amido modified up-conversion fluorescence nano material NaY
0.78F
4: Yb
0.20, Ho
0.02
(3) adopt single stage method to prepare the magnetic nanoparticle of surface amination.
In 30mL monoethylene glycol, add 6.5g1,6-hexane diamine, 2.0g anhydrous sodium acetate (CH
3COONa) and 1.0g Iron(III) chloride hexahydrate (FeCl
3.6H
2O).Be heated to 50 ℃ then, stir and form uniform colloidal solution.Gained solution is transferred in the 50mL band teflon-lined agitated reactor reaction 6h under 198 ℃ of parts.After taking out agitated reactor, at room temperature natural cooling is abandoned supernatant liquid in the still, and the lower black solid to beaker, utilizes the magnetic separated and collected with deionized water rinsing then.Under the ultrasound condition, use aqueous dispersion again, the magnetic separated and collected is used washing with alcohol 2 times again by top method then, and the gained black solid is dry 5-10h under 50 ℃ of conditions.
(4) magnetic nanoparticle and anti--CEA first antibody (capture antibody) combines.
10mg is shown amidized Fe
3O
4Magnetic nanoparticle joins 5mL 10mM PBS damping fluid (pH7.4), and ultrasonic dispersion added 1.25mL 25% glutaraldehyde and 100mg NaBH after 20 minutes
4Mixed solution is at room temperature slowly rocked 1h, and reaction finishes the back magnetic Nano material and separates through magnetic, and the pressed powder that obtains is washed 3 times with thorough removal glutaraldehyde with 10mM PBS.Powder is again with 10mM PBS dissolving, and ultrasonic scattering adds 5mL 0.1mgmL
-1Anti--CEA one anti-, this solution at room temperature slowly rocks 6h, reaction finishes the back cleans repeatedly, abandons supernatant.The gained sediment, promptly magnetic nanoparticle/anti--CEA one anti-compound (capture probe) is resuspended with 5mL0.01M PBS damping fluid, under 4 ℃ of conditions, preserves subsequent use.
(5) last conversion nano particle and anti--CEA SA (detection antibody) combines.
The up-conversion fluorescence nano material that 10mg is amido modified joins 5mL 10mM PBS damping fluid (pH7.4), and ultrasonic dispersion added 1.25mL 25% glutaraldehyde and 100mg NaBH after 20 minutes
4Mixed solution is at room temperature slowly rocked 1h, and reaction finishes the back magnetic Nano material and separates through magnetic, and the pressed powder that obtains is washed 3 times with thorough removal glutaraldehyde with 10mM PBS.Powder is again with 10mM PBS dissolving, and ultrasonic scattering adds 5mL 0.15mgmL
-1Anti--CEA two anti-, this solution at room temperature slowly rocks 6h, reaction finishes the back cleans repeatedly, abandons supernatant.The gained sediment is promptly gone up conversion nano particle/anti--CEA two anti-compounds (capture probe), and is resuspended with 5mL 10mM PBS damping fluid, under 4 ℃ of conditions, preserves subsequent use.
Embodiment 2: the foundation of typical curve
With PBS damping fluid preparation variable concentrations (1 * 10
-12, 5 * 10
-12, 1 * 10
-11, 5 * 10
-11, 1 * 10
-10, 5 * 10
-10, 1 * 10
-9, 5 * 10
-9, 1 * 10
-8GmL
-1) the CEA standard items, get 100 μ L variable concentrations CEA standard items and equal-volume 0.1mgmL respectively
-1The magnetic nanoparticle of CEA antibody labeling fully mixes, and magnetic separates removed supernatant in 5 minutes, and resuspended magnetic nanoparticle also separates 3 times with the unconjugated CEA molecule of thorough removal with the washing of PBS damping fluid, magnetic.Add to go up conversion nano particle/anti--CEA two anti-compounds again, fully mixing and hatch 30min after divide the abandonment supernatant through magnetic again, clean 3 times to guarantee thoroughly to remove unconjugated detector probe.With the resuspended deposition of 100 μ L0.01M PBS damping fluids, under 980nm laser excitation, obtain the up-conversion fluorescence signal at 542nm place, fluorescence signal (I) progressively strengthens along with the increase of CEA concentration.Set up typical curve according to fluorescence intensity with corresponding CEA standard items concentration.Experimental result shows that this detection method is 2.5 * 10
-12-1 * 10
-8GmL
-1Obtain the good linear relation in the CEA concentration interval.
Embodiment 3:
The detection of CEA in the blood serum sample
Get 7 routine clinical serum specimens and CEA antigen standard items, serum specimen is with 10 times of PBS damping fluid dilutions, and the CEA standard items are made into variable concentrations (1 * 10 with the PBS damping fluid
-12, 5 * 10
-12, 1 * 10
-11, 5 * 10
-11, 1 * 10
-10, 5 * 10
-10, 1 * 10
-9, 5 * 10
-9, 1 * 10
-8GmL
-1).Utilize this kit and electrochemiluminescence kit (Roche diagnostic companies product) to measure the content of CEA in these samples respectively; The result sees table one, and the data that obtain are carried out correlativity relatively, and both do not have significant difference; The result has very high correlation (R=0.9929, P＜0.0001).Explain that CEA detection kit of the present invention compares with the similar products of external well-known manufacturer production, technical indicator is roughly suitable, but this kit detects fast and reliable, and sensitivity is higher, and stability is better, is suitable for the detection of CEA in the clinical blood serum sample.
Table one: clinical blood serum sample CEA detects, the inventive method and electrochemical luminescence method contrast
Annotate: NE is the PBS damping fluid, and n.d. is not for detecting.
Embodiment 4: the detection of CEA and recovery of standard addition experiment among the human serum sample
From 9 groups of CEA concentration datas that embodiment 1 obtains, select 2 groups as background values, then choose 0.5ngmL
-1, 1ngmL
-1, 5ngmL
-1The CEA standard items of three kinds of variable concentrations add to respectively in the determinand, utilize this kit to detect the wherein content of CEA once more equally, obtain detected value.Recovery %=(detected value-background values)/addition * 100%.From the result of table two, the recovery explains that 91.80%～114.10% the present invention is stable, and sensitivity accurately, is suitable for the detection of CEA in the blood serum sample.
Table two: the detection of CEA and recovery of standard addition among the human serum sample
The present invention combines embodiment to describe.Should be understood that these embodiment only are used for illustration purpose, and be not used in restriction scope of the present invention.Should be understood that in addition that after having read foregoing of the present invention those skilled in the art can do various changes and modification to the present invention, these equivalent form of values fall within the application's book appended claims institute restricted portion equally.
Claims (7)
1. a carcinomebryonic antigen (CEA) detection kit is characterized in that said kit comprises:
Magnetic Nano material as the capture antibody functionalization of antigen capture and magnetic separation agent; Finishing detect the fluorescence probe that the up-conversion fluorescence nano material of antibody constitutes; Positive control; Negative control; The antigen standard items.
2. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that: said antigen capture and magnetic separation agent are Fe
3O
4Magnetic Nano microsphere is assembled the magnetic Nano material of the CEA capture antibody functionalization that obtains with anti--CEA first antibody covalent bond.
3. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that: said fluorescence probe is NaY
0.78F
4: Yb
0.20, Ho
0.02Up-conversion fluorescence nano material and anti--CEA SA combine finishing that assembling obtains CEA detect the up-conversion fluorescence nano material of antibody.
4. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that: the fluorescence probe of said detection antibody labeling is with 980nm induced with laser NaY
0.78F
4: Yb
0.20, Ho
0.02Up-conversion fluorescence obtains detection signal.
5. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that, anti--CEA first antibody concentration is 0.1mgmL
-1, itself and Fe
3O
4The mass ratio of magnetic Nano microsphere is 1: 100.
6. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that, anti--CEA SA concentration is 0.15mgmL
-1, with NaY
0.78F
4: Yb
0.20, Ho
0.02The mass ratio of up-conversion fluorescence nano material is 3: 200.
7. carcinomebryonic antigen detection kit as claimed in claim 1 is characterized in that: antigen standard items CEA antigen freeze-dried powder; Positive control is the human serum that contains finite concentration CEA; Negative control is the PBS damping fluid, pH=7.4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210062667.5A CN102636649B (en) | 2012-03-04 | 2012-03-04 | Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210062667.5A CN102636649B (en) | 2012-03-04 | 2012-03-04 | Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102636649A true CN102636649A (en) | 2012-08-15 |
| CN102636649B CN102636649B (en) | 2014-11-26 |
Family
ID=46621118
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210062667.5A Expired - Fee Related CN102636649B (en) | 2012-03-04 | 2012-03-04 | Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102636649B (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103808923A (en) * | 2013-11-25 | 2014-05-21 | 珠海迪尔生物工程有限公司 | Movable type magnetic separation fluorescence immunoassay detection and analysis method and movable type magnetic separation fluorescence immunoassay detection and analysis device |
| CN104422770A (en) * | 2013-08-23 | 2015-03-18 | 上海凯璟生物科技有限公司 | Up-conversion luminescence-based chip detection system and reagent for gynecological tumor markers CA125, CA153, SCCA, HE4 |
| CN105241869A (en) * | 2015-09-29 | 2016-01-13 | 江南大学 | Bisphenol A electrochemiluminescent aptamer sensor based on upper conversion nano material |
| CN105424662A (en) * | 2015-11-24 | 2016-03-23 | 江南大学 | Fluorescent biological detection system |
| CN105651989A (en) * | 2016-01-28 | 2016-06-08 | 天津科技大学 | Blue-light-emitting up-conversion material and preparation and application thereof |
| CN107525937A (en) * | 2017-08-25 | 2017-12-29 | 苏州优函信息科技有限公司 | Based on up-conversion luminescence label, protein chip and detection method |
| CN111017978A (en) * | 2019-12-27 | 2020-04-17 | 江西鑫泰功能材料科技有限公司 | Preparation method of nano holmium oxide for dielectric ceramic capacitor |
| CN111465856A (en) * | 2017-12-11 | 2020-07-28 | 株式会社德行健 | Composition for tuberculosis diagnosis and method for tuberculosis diagnosis based on change in optical properties |
| CN111505264A (en) * | 2019-01-30 | 2020-08-07 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN112462056A (en) * | 2020-11-19 | 2021-03-09 | 武汉大学 | Urine detection platform for detecting bacteria in urine on site and use method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004333168A (en) * | 2003-04-30 | 2004-11-25 | Fuji Photo Film Co Ltd | Biochemical analytical method using storage phosphor |
| CN1580775A (en) * | 2003-08-08 | 2005-02-16 | 清华大学 | Nano fluorescent magnetic particle and its preparing method |
| CN101776683A (en) * | 2010-01-06 | 2010-07-14 | 东华大学 | Luminescent and magnetic nano material-based method for detecting carcinoembryonic antigen |
| CN102023147B (en) * | 2010-09-29 | 2012-02-22 | 江南大学 | Method for detecting ochratoxin A by magnetic separation of adapter-functionalized magnetic nano material and marking of up-conversion fluorescent nano material |
-
2012
- 2012-03-04 CN CN201210062667.5A patent/CN102636649B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004333168A (en) * | 2003-04-30 | 2004-11-25 | Fuji Photo Film Co Ltd | Biochemical analytical method using storage phosphor |
| CN1580775A (en) * | 2003-08-08 | 2005-02-16 | 清华大学 | Nano fluorescent magnetic particle and its preparing method |
| CN101776683A (en) * | 2010-01-06 | 2010-07-14 | 东华大学 | Luminescent and magnetic nano material-based method for detecting carcinoembryonic antigen |
| CN102023147B (en) * | 2010-09-29 | 2012-02-22 | 江南大学 | Method for detecting ochratoxin A by magnetic separation of adapter-functionalized magnetic nano material and marking of up-conversion fluorescent nano material |
Non-Patent Citations (1)
| Title |
|---|
| MENG WANG，ET AL.: "Immunolabeling and NIR-Excited Fluorescent Imaging of HeLa Cells by Using NaYF4:Yb,Er Upconversion Nanoparticles", 《ACS NANO》 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104422770A (en) * | 2013-08-23 | 2015-03-18 | 上海凯璟生物科技有限公司 | Up-conversion luminescence-based chip detection system and reagent for gynecological tumor markers CA125, CA153, SCCA, HE4 |
| CN103808923A (en) * | 2013-11-25 | 2014-05-21 | 珠海迪尔生物工程有限公司 | Movable type magnetic separation fluorescence immunoassay detection and analysis method and movable type magnetic separation fluorescence immunoassay detection and analysis device |
| CN105241869A (en) * | 2015-09-29 | 2016-01-13 | 江南大学 | Bisphenol A electrochemiluminescent aptamer sensor based on upper conversion nano material |
| CN105424662A (en) * | 2015-11-24 | 2016-03-23 | 江南大学 | Fluorescent biological detection system |
| WO2017088214A1 (en) * | 2015-11-24 | 2017-06-01 | 江南大学 | Fluorescence-based biological detection system |
| CN105651989A (en) * | 2016-01-28 | 2016-06-08 | 天津科技大学 | Blue-light-emitting up-conversion material and preparation and application thereof |
| CN107525937A (en) * | 2017-08-25 | 2017-12-29 | 苏州优函信息科技有限公司 | Based on up-conversion luminescence label, protein chip and detection method |
| CN111465856A (en) * | 2017-12-11 | 2020-07-28 | 株式会社德行健 | Composition for tuberculosis diagnosis and method for tuberculosis diagnosis based on change in optical properties |
| CN111505264A (en) * | 2019-01-30 | 2020-08-07 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN111505264B (en) * | 2019-01-30 | 2024-05-17 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN111017978A (en) * | 2019-12-27 | 2020-04-17 | 江西鑫泰功能材料科技有限公司 | Preparation method of nano holmium oxide for dielectric ceramic capacitor |
| CN112462056A (en) * | 2020-11-19 | 2021-03-09 | 武汉大学 | Urine detection platform for detecting bacteria in urine on site and use method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102636649B (en) | 2014-11-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102636649B (en) | Kit for detecting carcinoembryonic antigen based on antibody functionalized magnetic nanometer material and up-conversion fluorescence nanometer material | |
| Wu et al. | Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B1 and ochratoxin A using upconversion nanoparticles as multicolor labels | |
| CN103245656B (en) | Preparation and application of alpha fetoprotein and carcino-embryonic antigen electrochemiluminescence sensor | |
| Liu et al. | Core–shell Fe3O4–Au magnetic nanoparticles based nonenzymatic ultrasensitive electrochemiluminescence immunosensor using quantum dots functionalized graphene sheet as labels | |
| CN102023147A (en) | Method for detecting ochratoxin A by magnetic separation of aptamer-functionalized magnetic nano material and marking of up-conversion fluorescent nano material | |
| CN101706498B (en) | Preparing method of electrochemiluminescence immunosensor | |
| CN102914646B (en) | Homogeneous phase multi-component immunoassay method based on surface plasma coupling effect | |
| CN103411948A (en) | SERS (Surface Enhanced Raman Spectroscopy) method for detecting alpha fetal protein based on probe-marked immunogold and magnetic composite materials | |
| CN105301241B (en) | Preparing method and application of sandwich type electrochemical luminescence immunosensor for detecting tumor marker | |
| CN109839501B (en) | Electrochemiluminescence immunosensor for measuring circulating tumor cells and preparation method and application thereof | |
| CN105115961B (en) | A kind of preparation method of the electrochemical luminescence sensor of nano composite material | |
| CN110687182A (en) | Preparation method of electrochemical immunosensor for detecting prostate specific antigen | |
| CN104880456A (en) | Preparation method and application of electrochemiluminescence immunosensor constructed on basis of GO/MWCNTs-COOH/Au @ CeO2 | |
| CN104568923A (en) | Method and kit for detecting circulating tumor cell antigens in peripheral blood through electrochemical luminescence detection | |
| Li et al. | Near-infrared photoactive Yb-MOF functionalized with a large conjugate ionic liquid: synthesis and application for photoelectrochemical immunosensing of carcinoma embryonic antigen | |
| CN107356570B (en) | Solid-state up-conversion fluorescent probe and preparation method and application thereof | |
| CN105203524A (en) | Method based on aptamer recognition surface enhanced Raman spectroscopy for detecting salmonella in food | |
| CN105954339A (en) | Preparation method and application of sandwich type immunosensor based on CeO2@Cu2O/Au@Pt | |
| CN109613244B (en) | Preparation method and application of Ag @ Pt-CuS labeled immunosensor | |
| Zhong et al. | Dual-wavelength responsive photoelectrochemical aptasensor based on ionic liquid functionalized Zn-MOFs and noble metal nanoparticles for the simultaneous detection of multiple tumor markers | |
| Song et al. | NIR-II fluorescent nanoprobe-labeled lateral flow biosensing platform: A high-performance point-of-care testing for carcinoembryonic antigen | |
| He et al. | A paper-supported sandwich immunosensor based on upconversion luminescence resonance energy transfer for the visual and quantitative determination of a cancer biomarker in human serum | |
| Lin et al. | A new dual immunoassay for tumor markers based on chemiluminescence signal amplification by magnetic mesoporous silica and enzyme modified gold nanoparticles | |
| Zhang et al. | Enhanced electrochemiluminescence of CdS quantum dots capped with mercaptopropionic acid activated by EDC for Zika virus detection | |
| CN103033618A (en) | Novel synthesis method of polyfluorene derivative and application in immunosensor for detecting tumor marker |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141126 Termination date: 20150304 |
|
| EXPY | Termination of patent right or utility model |