CN105717286B - A method of taxanes substance is fixed on solid phase carrier - Google Patents
A method of taxanes substance is fixed on solid phase carrier Download PDFInfo
- Publication number
- CN105717286B CN105717286B CN201410734480.4A CN201410734480A CN105717286B CN 105717286 B CN105717286 B CN 105717286B CN 201410734480 A CN201410734480 A CN 201410734480A CN 105717286 B CN105717286 B CN 105717286B
- Authority
- CN
- China
- Prior art keywords
- taxanes
- solid phase
- substance
- taxol
- phase carrier
- 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
Links
Abstract
The invention discloses a kind of to be fixed on taxanes substance method on solid phase carrier, the surface of the solid phase carrier is combined with maleic anhydride, the method is to be reacted with the maleic anhydride by taxanes substance, to which the taxanes substance to be fixed on the solid phase carrier.It, can be with the concentration of taxanes substance in determination sample according to Reverse transcriptase enzyme-linked immunization principle using the specific antibody of taxanes substance after taxanes substance is fixed on solid phase carrier by method through the invention.The present invention method compared with existing method have many advantages, such as it is more convenient, quick, sensitive, practical in the detection of taxanes substance.
Description
Technical field
The present invention relates to enzyme linked immunosorbent detection fields, and taxanes substance is fixed on solid phase carrier more particularly to a kind of
On method.
Background technology
Taxol (taxol) is a kind of tetracyclic diterpene compound, is cell mitogen inhibitor, is used for cancer and controls
It treats.Microtubule inhibitors in taxol category mitosis, taxol have the function of polymerizeing and stablizing intracellular canaliculus, cause fast
The tumour cell of speed division is firmly fixed in mitotic stages, is kept micro-pipe no longer separated, cell can be blocked in the cell cycle
G2 and M phases, so that cancer cell duplication is blocked and dead.In treatment of cancer, the content of taxol is to treatment in patient tissue
Effect is very crucial, it usually needs maintains lower level.Because for cell, paclitaxel concentration, which crosses conference, leads to normal cell
Mortality.Therefore, the content of taxol and metabolism need a kind of quick, sensitive detection side in frequent monitoring patient tissue
Method.
Currently, the detection method of taxol has uv detection method, high performance liquid chromatography and Reverse transcriptase enzyme linked immunological
Method (Competitive Inhibition Enzyme Immunoassay, CIEIA) etc., uv detection method and high-efficient liquid phase color
Spectrometry needs a large amount of acquisition samples, and detects compared with slow, cost is excessively high；CIEIA is to utilize enzyme linked immunological principle, anti-by developing the color
It answers, without further purification, the method that quickly detection slightly carries taxol in mixture.Jaziri etc. can detect plant group with this method
Knit and callus in 1-100ng contents the taxol (1991.Enzyme-linked such as Jaziri immunosorbent
assay for the detection and the semi-quantitative determination of taxane
diterpenoids related to taxol in Taxus sp.and tissue cultures.J Pharm
Belg.1991Mar-Apr；46(2):93-9).Using this method, Grothaus etc. can detect 0.3ng/ml in blood plasma
Taxol (the 1993.An enzyme immunoassay for the determination of such as Grothaus taxol
and taxanes in Taxus sp.tissues and human plasma.J Immunol Methods.1993Jan
14；158(1):5-15.).
In the scheme that existing CIEIA detects taxol, as shown in Figure 1, taxol is generally by pyridine solution
It is reacted with succinic anhydride, generates 2- taxol succinyl monoesters, bovine serum albumin(BSA) (BSA) solution and 25% dioxy is then added
Clustered water solution (v/v) makes 2- taxol succinyl monoesters pass through amido bond with the amino of BSA and is combined to form BSA coupled products,
It is fixed on microwell plate 1 by the suction-operated of BSA, then passes through the content of taxol in CIEIA determination samples again.Due to inciting somebody to action
Taxol is fixed on that this method step on microwell plate is more, and efficiency is low, the time is long, cannot meet often monitoring patient group well
Knit the demand of the content of middle taxol.
Invention content
In order to make up above-mentioned the deficiencies in the prior art, present invention proposition is a kind of to be fixed on solid phase carrier by taxanes substance
On method.
The technical problem of the present invention is resolved by technical solution below：
A method of taxanes substance is fixed on solid phase carrier, the surface of the solid phase carrier is combined with Malaysia
Acid anhydrides, the method is to be reacted with the maleic anhydride by taxanes substance, to fix the taxanes substance
On the solid phase carrier.
Preferably, the taxanes substance reacts with the maleic anhydride and includes the following steps：
(1) solution is prepared, contains the taxanes substance and pyridine, in every milliliter of pyridine, institute in the solution
The content for stating taxanes substance is 100-1000 micrograms；
(2) solid phase carrier is added in the solution, in the case where reaction temperature is 25-65 DEG C, the taxanes substance
2 '-hydroxyls reacted with maleic anhydride at least 8 hours, formed 2 '-maleic acid taxanes esters.
Preferably, the taxanes substance reacts with the maleic anhydride and includes the following steps：
(1) solution is prepared, the taxanes substance and pyridine solution are contained in the solution, the pyridine solution includes pyrrole
The volume ratio of pyridine and the solvent that can be dissolved each other with the pyridine, the pyridine and solvent is more than or equal to 1% and is less than 100%, every
In the milliliter pyridine solution, the content of the taxanes substance is 100-1000 micrograms；
(2) solid phase carrier is added in the solution, in the case where reaction temperature is 25-65 DEG C, the taxanes substance
2 '-hydroxyls reacted with maleic anhydride at least 8 hours, formed 2 '-maleic acid taxanes esters.
In pyridine, the maleic anhydride of 2 ' more active-OH and surface of solid phase carriers reacts taxol, and 7 '-OH by
It is more inactive in the space structure the problem of, therefore react and generate 2 '-maleic acid taxanes esters.
Preferably, the content of the taxanes substance is 200-300 micrograms.
Preferably, in step (2), reaction temperature is 50-60 DEG C.
Preferably, the volume ratio of the pyridine and solvent is 1-5%.
Preferably, the reaction time in step (2) is 30-40 hours.
Preferably, the solvent in step (1) is water, PBS buffer solution, methanol, mineral oil, formaldehyde, dimethyl sulfoxide (DMSO)
At least one of with n-hexane.
Preferably, the taxanes substance has the following structure formula：
Work as R1=C6H5, R2=CH3It is taxol when CO；Work as R1=(CH3)3CO, R2It is Docetaxel when=H.
A kind of intermediate for detecting taxanes substance, including solid phase carrier and taxanes substance, the solid phase
The surface of carrier is combined with maleic anhydride, and the taxanes substance is described by reacting to be incorporated in the maleic anhydride
On solid phase carrier.
Advantages of the present invention：The method of the present invention is to bind directly taxol using the solid phase carrier activated through maleic anhydride
Substance, so as to be directly entered subsequent CIEIA detecting steps, reactions steps of this method is few, and interference impurity is few, and reaction is special
One property is strong compared with existing method have many advantages, such as it is more convenient, quick, sensitive, practical in the detection of taxanes substance.
Description of the drawings
Fig. 1 is the reaction schematic diagram that taxol in the prior art is fixed to by BSA coupled products in micropore plate surface；
Fig. 2 is the reaction schematic diagram that the taxol in the embodiment of the present invention 1 is fixed on orifice plate；
Fig. 3 is the principle schematic of the content of taxol in CIEIA detection samples to be tested；
Fig. 4 is the absorption curves measured at 450 nm in the embodiment of the present invention 2；
Fig. 5 is existed according to paclitaxel standard object obtained under the linear 0.05-5ng/mL concentration ranges in Fig. 4
The working curve in the sections 0.05-5ng/mL.
Specific implementation mode
Below against attached drawing and in conjunction with preferred embodiment, the invention will be further described.
The present invention provides a kind of method being fixed on taxanes substance on solid phase carrier, in a kind of specific implementation mode
In, the surface of the solid phase carrier is combined with maleic anhydride, and the method is to pass through taxanes substance and the maleic anhydride
Reaction, to which the taxanes substance to be fixed on the solid phase carrier.
More preferably, the method is to react to be formed with the maleic anhydride by the 2 '-hydroxyls by taxanes substance
2 '-maleic acid taxanes esters.
In some preferably embodiment, the taxanes substance is reacted with the maleic anhydride including following step
Suddenly：
(1) solution is prepared, contains the taxanes substance and pyridine, in every milliliter of pyridine, institute in the solution
The content for stating taxanes substance is 100-1000 micrograms；
(2) solid phase carrier is added in the solution, in the case where reaction temperature is 25-65 DEG C, the taxanes substance
2 '-hydroxyls reacted with maleic anhydride at least 8 hours, formed 2 '-maleic acid taxanes esters.
In other preferably embodiment, the taxanes substance is reacted with the maleic anhydride including following step
Suddenly：
(1) solution is prepared, the taxanes substance and pyridine solution are contained in the solution, the pyridine solution includes pyrrole
The volume ratio of pyridine and the solvent that can be dissolved each other with the pyridine, the pyridine and solvent is more than or equal to 1% and is less than 100%, every
In the milliliter pyridine solution, the content of the taxanes substance is 100-1000 micrograms；
(2) solid phase carrier is added in the solution, in the case where reaction temperature is 25-65 DEG C, the taxanes substance
2 '-hydroxyls reacted with maleic anhydride at least 8 hours, formed 2 '-maleic acid taxanes esters.
Since the pyridine of excessive concentrations has the solid support material of organic plastics matrix certain corrosiveness, to shadow
Following reaction is rung, therefore before solid phase carrier is added, adds another solvent that can be dissolved each other with pyridine to reduce pyridine
Concentration when the volume ratio of pyridine and solvent is 1-5%, can be made reaction result fine and prevent pyridine to solid by experiment
The corrosion of phase carrier.
In the above preferred embodiment, the shadow of taxanes substance and maleic anhydride reacted also by reaction temperature
It rings, reaction bonded effect of the experiment test at 25 DEG C, 37 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, the temperature such as 65 DEG C is found
When reaction temperature is 50-60 DEG C, effect is more excellent.
Similarly, the reaction time also influences the efficiency of reaction, experiment test 8 hours, 16 hours, 24 hours, 32 hours,
Reaction in 40 hours and 48 hours finds that the efficiency that the reaction response time is 30-40 hours is higher., it was also found that Japanese yew in experiment
The content of alcohols material is more excellent when being 200-300 micrograms.
Wherein preferably：
The solvent is at least one in water, PBS buffer solution, methanol, mineral oil, formaldehyde, dimethyl sulfoxide (DMSO) and n-hexane
Kind.
The taxanes substance has the following structure formula：
Work as R1=C6H5, R2=CH3It is taxol when CO；Work as R1=(CH3)3CO, R2It is Docetaxel when=H.
Solid phase carrier can be commercially available the microwell plate for ELISA activated with Maleic Anhydride Surface, such as match silent
Fly Pierce Amine-binding, the Maleic Anhydride of (Thermo Fisher Scientific Inc.) production
96 orifice plates of Activated；Can also be conventional microwell plate, such as polyvinyl chloride, polystyrene, poly- propionamide and cellulose
Then the microwell plate of material is activated on the surface of microwell plate with maleic anhydride again so that maleic anhydride combines and is connected to microwell plate
On.Preferably it is 96 holes using trade name Pierce Amine-binding, Maleic Anhydride Activated
Plate.
The present invention also provides a kind of intermediates for detecting taxanes substance comprising solid phase carrier and taxanes
The surface of substance, the solid phase carrier is combined with maleic anhydride, and the taxanes substance with the maleic anhydride by reacting
To be fixed on the solid phase carrier.
Below by taking taxol as an example, by specific embodiment, the present invention will be further elaborated.
Embodiment 1
Taxol and 96 holes solid phase carrier Pierce Amine-binding, Maleic Anhydride Activated
The combination on plate (hereinafter referred to as orifice plate 2) surface, reaction schematic diagram are as shown in Figure 2.
It is as follows：
(1) solution is prepared, contains paclitaxel material, pyridine and solvent in the solution, the volume ratio of pyridine and solvent is
2.3%, in the liquid that every milliliter is made of pyridine and solvent, the content of paclitaxel material is 233 micrograms.
Specially：Weigh 10 milligrams of taxols, be dissolved in 1 milliliter of pyridine, then take 105 microlitres therein it is micro- with 4395
Rise formaldehyde (in other embodiments, or water, PBS buffer solution, methanol, mineral oil, formaldehyde, dimethyl sulfoxide (DMSO) or just oneself
Alkane or these one or more of mixing), obtain paclitaxel concentration be 233 mcg/mls 2.3% pyridine/formaldehyde it is molten
Liquid.
(2) solid phase carrier is added in above-mentioned solution, at 55 DEG C, the taxol reacts 40 hours with maleic anhydride, shape
At 2 '-maleic acid Japanese yew alcohol esters, to which the taxol is fixed on the solid phase carrier.
Specially：
Solution in the step of taking 150 microlitres (1) is added in each micropore of orifice plate 2, is enclosed in closed container, 55
40h is reacted under the conditions of DEG C, to be fixed on taxol in the micropore of orifice plate 2.
Measurement for content of taxol in the drafting of the working curve in the following embodiments of progress and sample is tested, in step
(2) after reacting, the reaction solution in micropore is outwelled, (buffer solution of PBS and nonionic surfactant composition may be used after washing
Washed, in this example three times using 1*PBST (PBS adds 0.05wt% polysorbas20s) washings) Japanese yew can be made by CIEIA
The content of taxol in the working curve or determination sample of alcohol reference substance.
The principle of the content of taxol is as follows in the working curve or determination sample of CIEIA making paclitaxel standard objects：
The taxol being fixed on microwell plate can be combined with taxol specific antibody, then pass through the coupling on secondary antibody
Marker enzyme (such as horseradish peroxidase) so that colorless substrate (such as TMB) is generated colored compound by enzyme-linked chromogenic reaction
(TMBO), in a certain range, the concentration of the taxol of color and micropore the plate surface combination of the reaction is linear, therefore
The working curve of reference substance can be made by the light absorption of measurement reaction solution, carry out determining the dense of tested object for calculating
Degree.
CIEIA detects the principle of the content of taxol in sample to be tested as shown in figure 3, being specially：In CIEIA, micro-
In the case that paclitaxel concentration is certain on orifice plate, by the way that taxol specific antibody is added simultaneously on microwell plate and contains unknown quantity
The sample to be tested of taxol so that the taxol competitive binding taxol specificity on microwell plate in taxol and sample to be tested
Antibody, the content of taxol in sample to be tested is higher, and competition is stronger, and is fixed on the taxol of the combination of the taxol on microwell plate
Specific antibody quantity is fewer, by washing away the taxol specific antibody and sample to be tested that are not associated in microwell plate
In taxol and taxol specific antibody combination product, it is anti-that enzyme-linked colour developing as described above is then carried out in micropore
It answers, the content of taxol in shade and sample to be tested is inversely proportional, and is calculated i.e. by the working curve of obtained reference substance
The content of taxol in sample to be tested can be measured.
Embodiment 2
The working curve of paclitaxel standard object is obtained using CIEIA, is as follows：
1) 350 microlitres of sealers are added (using containing 1wt% in this example in being fixed in the micropore of taxol for embodiment 1
The PBST solution of ovalbumin is as sealer), capping is carried out at 20-30 DEG C 6 hours.
2) it prepares taxol specific antibody with the PBST solution of the ovalbumin containing 2wt% and (uses rabbit-anti Japanese yew in this example
Alcohol polyclonal antibody (Abcam, ab26953)) and taxol respectively containing following concentration：0,0.04,0.4,4,40,400 nanograms/
Microlitre paclitaxel standard liquid, wherein taxol specific antibody and the volume ratio of the PBST solution of the ovalbumin containing 2wt% are
1:250。
3) take the above-mentioned paclitaxel standard liquid of 150 microlitres of taxols containing various concentration that the micropore of step 1) is added respectively
In, carry out the competitive inhibition reaction of taxol and taxol specific antibody for 24 hours at 4 DEG C.Reaction solution is outwelled after the completion of reaction,
It is used in combination 350 microlitres of PBST solution to wash 3 times.
4) 1 is prepared with the PBST solution of the ovalbumin containing 1wt%:5000 (have been coupled the secondary antibody of enzyme marker and have contained 1% ovum
The volume ratio of albuminised PBST solution) secondary antibody of enzyme marker has been coupled (using being coupled horseradish peroxidase in this example
Goat-anti rabbit secondary antibody), and take 150 microlitres to be added in the micropore in step 3), it is reacted 2 hours at 20-30 DEG C.Reaction solution is outwelled,
It is used in combination 350 microlitres of PBST solution to wash 3 times.Then add 150 microlitres of TMB (3,3', 5,5'- tetramethyl benzidine) bottoms into micropore
Object reaction solution reacts 15-30 minutes at 20-30 DEG C, or to there is apparent color to occur, the sulfuric acid that 75 microlitres of 2M are added terminates
Reaction measures light absorption and obtains standard curve as shown in Figure 4, at 450 nm wherein in 0.05-5ng/mL concentration ranges at line
Property, the working curve of paclitaxel standard object of the taxol in the sections 0.05-5ng/mL is drawn according to the linear region, is such as schemed
It shown in 5, is shown in Fig. 5, the relationship between paclitaxel concentration and absorbance meets y=-43.522Ln (x)+25.567, and linear
Regression coefficient is close to 1 (R2=0.9938), illustrate that the data that the section measures are reliable.
Embodiment 3
Using the content of taxol in CIEIA determination samples, it is as follows：
S1, the fresh Chinese yew needle of acquisition, are cleaned with clear water, in being dried to constant weight, with pulverizer powder in 65 DEG C of baking ovens
It is broken, the raw material 2g after crushing accurately is weighed, is set in 50mL ground conical flask with stopper, ethyl alcohol 20mL, ultrasonic extraction 30min is added,
Supernatant, precipitation is stayed to be added with 20mL ethyl alcohol, repetitive operation.Merge supernatant, filtering and removing slag, filtrate must soak in 30 DEG C of evaporated under reduced pressure
Cream, medicinal extract 20mL methanol dissolves, and the petroleum ether of 1 times of volume is added, and vibrates 30min, degreasing；Extract extraction raffinate chloroform
(volume ratio 2:1) 30min is vibrated, is faded, is not thorough and should be repeated if fading；Extraction extraction raffinate reduced pressure is evaporated (30 again
DEG C), paclitaxel extract is dissolved to obtain with 400 μ L methanol, for CIEIA test analysis.
S2, the PBST solution of above-mentioned paclitaxel extract ovalbumin containing 2wt% is diluted to 50,000 times, and is added 1:
More grams of the rabbit-anti taxol of 250 (volume ratios of rabbit-anti taxol polyclonal antibodies and the PBST solution of the ovalbumin containing 2wt%)
Grand antibody (Abcam, ab26953), as sample to be tested.
S3, the PBST that 350 microlitres of ovalbumins containing 1wt% are added in the micropore for combining taxol of embodiment 1 are molten
Liquid, carries out capping 6 hours at 20-30 DEG C.
S4, it takes the sample to be tested in 150 microlitres of step S2 to be added in the micropore in step S3, taxol is carried out at 4 DEG C
For 24 hours with the competitive inhibition reactions of rabbit-anti taxol polyclonal antibodies.Above-mentioned reaction solution is outwelled after the completion of reaction, is used in combination 350 microlitres
PBST solution wash 3 times.
S5,1 is prepared with the PBST solution of the ovalbumin containing 1wt%:5000 (have been coupled the goat-anti rabbit two of horseradish peroxidase
The anti-volume ratio with the PBST solution of the ovalbumin containing 1wt%) it has been coupled the goat-anti rabbit secondary antibody of horseradish peroxidase, and take 150
It microlitre is added in the micropore of step S4, is reacted 2 hours at 20-30 DEG C.Reaction solution is outwelled, 350 microlitres of PBST solution is used in combination to wash
It washs 3 times.It into micropore plus 150 microlitres of tmb substrate reaction solutions, is reacted 15-30 minutes at 20-30 DEG C, or to there is apparent face
Color occurs, and the sulfuric acid that 75 microlitres of 2M are added terminates reaction, measures light absorption at 450 nm, repeats the above steps three times, measure number
According to as shown in table 1 below.
Table 1:The light absorption of each sample that CIEIA is measured at 450 nm
| It compares (micropore does not combine taxol) | Blank control (0ng/mL) | Sample to be tested (unknown concentration) | |
| Repeat 1 | 0.9484 | 1.902 | 1.6993 |
| Repeat 2 | 0.9492 | 1.7348 | 1.6905 |
| Repeat 3 | 0.9422 | 1.7908 | 1.6717 |
| Average value | 0.9466 | 1.8092 | 1.6872 |
| Variance | 0.0038 | 0.0851 | 0.0141 |
S6, according to the working curve of the reference substance as shown in Figure 5 obtained in embodiment 2, calculate in sample to be tested purple
A concentration of 2.803 nanograms/milliliter of China fir alcohol, the paclitaxel concentration being equivalent in undiluted preceding paclitaxel extract are 140.1
Mcg/ml.
As a contrast, pass through UPLC (ultra performance liquid chromatography, Ultra Performance Liquid
Chromatography, UPLC) paclitaxel extract in step S1 is directly measured, measure a concentration of 151.2 microgram/
Milliliter, error 7.34%.This illustrates that CIEIA measurement has good reliability, but sensitiveer than the methods of UPLC (because of dilution
It still can detect after 50,000 times).Therefore, method of the invention has the characteristics that simple, convenient, fast, sensitive, can apply
In clinic, and too many sample need not be acquired from patient, so that it may with Japanese yew in tissue during monitoring chemotherapy
The variation of determining alcohol has good application value.
Above example is the explanation carried out by taking taxol as an example, and the method for each embodiment is to other taxanes objects
Matter, as Docetaxel is equally applicable.
The above content is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be said that
The specific implementation of the present invention is confined to these explanations.For those skilled in the art to which the present invention belongs, it is not taking off
Under the premise of from present inventive concept, several equivalent substitute or obvious modifications can also be made, and performance or use is identical, all answered
When being considered as belonging to protection scope of the present invention.
Claims (3)
1. a kind of method being fixed on taxanes substance on solid phase carrier, it is characterised in that：The surface of the solid phase carrier
It is combined with maleic anhydride, the method is to be reacted with the maleic anhydride by taxanes substance, thus by the taxol
Substance is fixed on the solid phase carrier；
The taxanes substance has the following structure formula：
Work as R1=C6H5, R2=CH3It is taxol when CO；Work as R1=(CH3)3CO, R2It is Docetaxel when=H；The taxanes
Substance is reacted with the maleic anhydride to be included the following steps：
（1）Prepare solution, contain the taxanes substance and pyridine solution in the solution, the pyridine solution include pyridine and
The volume ratio of the solvent that can be dissolved each other with the pyridine, the pyridine and solvent is 1-5%, in every milliliter of pyridine solution, institute
The content for stating taxanes substance is 200-300 micrograms；
（2）The solid phase carrier is added in the solution, in the case where reaction temperature is 50-60 °C, the 2 '-of the taxanes substance
Hydroxyl reacts at least 8 hours with maleic anhydride, forms 2 '-maleic acid taxanes esters.
2. taxanes substance to be fixed on to the method on solid phase carrier as described in claim 1, it is characterised in that：Step
（2）In reaction time be 30-40 hours.
3. taxanes substance to be fixed on to the method on solid phase carrier as described in claim 1, it is characterised in that：Step
（1）In the solvent be water, PBS buffer solution, methanol, mineral oil, formaldehyde, dimethyl sulfoxide (DMSO) and n-hexane at least one
Kind.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410734480.4A CN105717286B (en) | 2014-12-04 | 2014-12-04 | A method of taxanes substance is fixed on solid phase carrier |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410734480.4A CN105717286B (en) | 2014-12-04 | 2014-12-04 | A method of taxanes substance is fixed on solid phase carrier |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105717286A CN105717286A (en) | 2016-06-29 |
| CN105717286B true CN105717286B (en) | 2018-08-21 |
Family
ID=56143999
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410734480.4A Active CN105717286B (en) | 2014-12-04 | 2014-12-04 | A method of taxanes substance is fixed on solid phase carrier |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105717286B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103889592A (en) * | 2011-07-14 | 2014-06-25 | 表面创新有限公司 | A method of producing a functionalized surface and surfaces made thereby |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9149440B2 (en) * | 2003-09-02 | 2015-10-06 | University Of South Florida | Nanoparticles for drug-delivery |
| US9175015B2 (en) * | 2007-08-22 | 2015-11-03 | Colorado School Of Mines | Gold nanoparticle conjugates and uses thereof |
-
2014
- 2014-12-04 CN CN201410734480.4A patent/CN105717286B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103889592A (en) * | 2011-07-14 | 2014-06-25 | 表面创新有限公司 | A method of producing a functionalized surface and surfaces made thereby |
Non-Patent Citations (2)
| Title |
|---|
| MICHAEL BÖCHER 等.Direct Coupling of an Atrazine Analogue to Microtiter Plates for a Competitive Immunoassay for s-Triazines.《Food and Agricultural Immunology》.1994,第6卷(第2期), * |
| 抗紫杉醇单克隆抗体的制备及ELISA检测方法的建立;赵凯 等;《中国新药杂志》;20120228;第21卷(第4期);436-442 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105717286A (en) | 2016-06-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure | |
| Chen et al. | Rational hapten design to produce high-quality antibodies against carbamate pesticides and development of immunochromatographic assays for simultaneous pesticide screening | |
| Xu et al. | Establishment of magnetic beads-based enzyme immunoassay for detection of chloramphenicol in milk | |
| Lei et al. | Development of an ultrasensitive ic-ELISA and immunochromatographic strip assay for the simultaneous detection of florfenicol and thiamphenicol in eggs | |
| Liu et al. | A highly sensitive “turn-on” fluorescent probe with an aggregation-induced emission characteristic for quantitative detection of γ-globulin | |
| WO2018001309A1 (en) | Protein chip and reagent kit for detecting abnormal serum des-gamma-carboxy-prothrombin and manufacturing method thereof | |
| CN104655847A (en) | Enzyme linked immunosorbent assay kit (ELISA kit) for detecting adprin and detection method thereof | |
| CN101377505A (en) | Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof | |
| CN104447984A (en) | Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent | |
| Shen et al. | A monoclonal antibody-based time-resolved fluoroimmunoassay for chloramphenicol in shrimp and chicken muscle | |
| CN104897864B (en) | The enzyme linked immunological kit of detection amantadine and application thereof | |
| Sun et al. | A sensitive heterogeneous biotin–streptavidin enzyme-linked immunosorbent assay for the determination of di-(2-ethylhexyl) phthalate (DEHP) in beverages using a specific polyclonal antibody | |
| CN101776685A (en) | Enzyme linked immunosorbent assay kit for detecting trimethoprim medicament and application thereof | |
| Tao et al. | Development of competitive indirect ELISA for the detection of tetrodotoxin and a survey of the distribution of tetrodotoxin in the tissues of wild puffer fish in the waters of south-east China | |
| Zeravik et al. | Development of direct ELISA for the determination of 4-nonylphenol and octylphenol | |
| CN105403703B (en) | Detect enzyme linked immunological kit and its application of carbendazim | |
| Yuan et al. | Development of a sensitive indirect competitive enzyme-linked immunosorbent assay based on the monoclonal antibody for the detection of benzothiostrobin residue | |
| CN105717286B (en) | A method of taxanes substance is fixed on solid phase carrier | |
| Pongkitwitoon et al. | Development of an enzyme-linked immunosorbent assay to determine puerarin and its aglycone daidzein | |
| CN1687782A (en) | Analytical kit of enzyme linked immunosorbent assay for residual carbaryl | |
| CN104383889B (en) | A kind of Dihydrocapsaicin polyclonal antibody adsorbent, immune affinity column and its preparation method and application | |
| Zhuang et al. | Artificial antigen synthesis and the development of polyclonal antibody-based immunoassay for citreoviridin determination | |
| Kemp et al. | Enzyme‐linked immunosorbent assay of 3‐acetyldeoxynivalenol applied to rice | |
| Chen et al. | Real-time immuno-PCR assay for detecting PCBs in soil samples | |
| Lv et al. | Development of a gold nanoparticle-based lateral-flow strip for the detection of cannabidiol in functional beverages |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |