CN103018462A - Nano-zinc oxide-modified immune capillary as well as preparation method and application thereof - Google Patents
Nano-zinc oxide-modified immune capillary as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a nano-zinc oxide-modified immune capillary as well as a preparation method and application thereof. The immune capillary is formed by growing nano-zinc oxide on the inner surface of a glass capillary and combining a detection probe with the nano-zinc oxide. As the nano-zinc oxide has a huge specific surface area, the surface density of antibody protein can be effectively improved, and the nano-zinc oxide has unique physical property for amplifying fluorescent signals of fluorescence-marked protein; and the capillary can provide micro-channels needed by flowing detection and has optical transparency property, so that the quick and quantitative immune detection is achieved, and the capillary can be used for early screening of malignant tumors and food safety analysis.
Description
Technical field
The present invention relates to a kind of immunoassay device, particularly nano oxidized Zinc modified immune capillary also relates to the preparation method and application of this mao immune capillary.
Background technology
Immunoassay is by specific combination between the antigen-antibody, to various materials as: medicine, hormone, protein, microorganism etc. carry out quantitatively and detection method qualitatively, generally are used for the fields such as medical diagnosis on disease, environmental monitoring, food security.For example: adopt immunoassay that the specific tumors mark in the serum is detected, the method is a kind of effective, the noninvasive method of tumour being carried out early diagnosis and examination.At present, it is enzyme linked immunosorbent assay that immune analysis method mainly adopts, and the method length consuming time, expense is high, need large-scale instrument and skilled operation technology, is not suitable for fast, on-the site analysis and extensive immune examination.Some portable immunity analysis instruments are also arranged on the market in addition, such as immunity test strip etc., can provide quick on-the-spot immunoassay, but this para-immunity analyser can only carry out qualitative or half-quantitative detection, quantitative test can not be provided.
U.S. Omowunmi A. Sadik in 2006 and Jason Karasinski adopt glass capillary to prepare immune detection device (United States Patent:7708944), this immune detectors can be used in quantitative test, but should the immunity device sensitivity low, signal is all once poor, is difficult to be applied to actual immunoassay.
Summary of the invention
In view of this, one of purpose of the present invention is to provide nano oxidized Zinc modified immune capillary, can realize fast immune detection, and the energy quantitative test; Two of purpose of the present invention is to provide the preparation method of nano oxidized Zinc modified immune capillary, and the preparation method is simple, good reproducibility; Three of purpose of the present invention is to provide the application of nano oxidized Zinc modified immune capillary.
For achieving the above object, technical scheme is:
1. nano oxidized Zinc modified immune capillary, then described immune capillary is formed in conjunction with detector probe on nano zine oxide by glass capillary inside surface growing nano zinc oxide.
Preferably, described nano zine oxide is zinc oxide nano rod or zinc oxide nano-particle.
Preferably, described detector probe is for detecting the monoclonal antibody of tumor markers.
Preferred, described monoclonal antibody is anti-human epidermal growth factor acceptor 2 monoclonal antibody, anticancer embryonal antigen monoclonal antibody, anti-prostate-specific-antigen monoclonal antibody or anti-alpha-fetoprotein monoclonal antibody.
2. the preparation method of described immune capillary comprises the steps:
A. glass capillary is activated;
B. with the activation the glass capillary inside surface with nano oxidized Zinc modified, get nano oxidized Zinc modified kapillary;
C. with the nano oxidized Zinc modified capillary surface of step b gained in conjunction with detector probe, get nano oxidized Zinc modified immune capillary.
Preferably, described step a is to be that 1-10mM potassium permanganate soaked 15-30 minute with glass capillary concentration.
Preferably, described step b puts into the mixed solution that contains zinc nitrate, monoethanolamine and ammoniacal liquor with the activation glass capillary, growth is 30-60 minute in 65-95 ℃ of water-bath, the final concentration of zinc nitrate is 10-100mM in the described mixed solution, the final volume mark of monoethanolamine is 1-10%, and the final volume mark of ammoniacal liquor is 1-10%.
Preferably, described step c is with the nano oxidized Zinc modified kapillary deionized water rinsing of step b gained, inject volume fraction after dry and be alcohol solution dipping 1-3 hour of 3-glycidoxypropyl trimethoxysilane of 1-5%, with behind the alcohol flushing under 110 ℃ of conditions vacuum annealing 1-3 hour, the detector probe that adds again 50-300 μ g/mL, soaked 2-8 hour, and soaked with bovine serum albumin(BSA) at last.
Preferably, described step a and b replace with following steps: will soak 20-60 minute in the colloidal solution implantation glass kapillary of zinc oxide nano-particle, and then calcine 15-60 minute nano oxidized Zinc modified kapillary under 150-200 ℃ of condition.
3. the described nano oxidized Zinc modified application of immune capillary in the preparation immunity testing equipment.
Beneficial effect of the present invention is: the present invention adopts the method for wet-chemical, prepared nano oxidized Zinc modified immune capillary, wherein nano zine oxide has huge specific surface area, the superficial density of energy Effective Raise sessile antibody albumen, nano zine oxide has the unique physical character of the fluorescence signal that amplifies fluorescence labeling albumen simultaneously; Kapillary then can provide the required microchannel of flow detection and because of its optical clear character, can realize easily optical detection; Cooperate a simple flow pumps and fluoroscopic examination device, this nano oxidized Zinc modified kapillary immunity device can be realized fast, scene, sensitive immunoassay.
Nano oxidized Zinc modified immune capillary disclosed by the invention can also by the kapillary of simple serial or parallel connection for the different target thing, realize that the polycomponent high flux detects.Because its preparation cost is low, therefore be highly suitable for extensive examination and field quick detection, be expected to be widely applied at aspects such as disease early screening, Food Safety Analysis, environmental contaminants monitoring.
Description of drawings
Fig. 1 is the electron scanning micrograph that is grown in the zinc oxide nano rod of capillary tube inner wall.(a is for to observe electron micrograph under 2 μ m, inserting figure among the figure is nano oxidized Zinc modified kapillary (left side) and the optical photograph of common kapillary (right side); B is electron micrograph under 200nm; C is electron micrograph under 100nm; D is electron microscope picture under the 100nm).
Fig. 2 is the immune capillary schematic diagram that zinc oxide nano rod is modified.
Fig. 3 is that nano oxidized Zinc modified immune capillary detects the typical curve that obtains to tumor markers HER2.
Fig. 4 is that nano oxidized Zinc modified immune capillary detects the typical curve that obtains to tumor markers CEA.
Fig. 5 is the electron scanning micrograph of capillary tube inner wall growth of zinc oxide nano particle.
Fig. 6 is the immune capillary schematic diagram that zinc oxide nano-particle is modified.
Fig. 7 is that nano oxidized Zinc modified immune capillary detects the typical curve that obtains to tumor markers PSA.
Fig. 8 is that nano oxidized Zinc modified immune capillary detects fluorescence photo and the intensity line that obtains to tumor markers AFP.
Fig. 9 is that nano oxidized Zinc modified kapillary immunity device detects the typical curve that obtains to tumor markers AFP.
Schematic diagram and fluorescence intensity line that Figure 10 detects polycomponent tumor markers HER2, CEA, PSA and AFP for the nano oxidized Zinc modified kapillary immunity device that adopts series connection.
The schematic diagram of Figure 11 for adopting nano oxidized Zinc modified kapillary immunity device in parallel that polycomponent tumor markers HER2, CEA, PSA and AFP are detected.
Embodiment
In order to make the purpose, technical solutions and advantages of the present invention clearer, the below is described in detail the preferred embodiments of the present invention.The experimental technique of unreceipted actual conditions in the preferred embodiment usually according to normal condition, or carries out according to the condition that manufacturer advises.
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) getting concentration is that 5 mM newly prepare liquor potassic permanganate implantation glass kapillary, soaks 20 minutes activation glass capillaries; Then use deionized water rinsing, then put into the 50 mM zinc nitrate solutions that contain 5% (v/v) monoethanolamine and 5% (v/v) ammoniacal liquor, growth is 50 minutes in 80 ℃ of water-baths, form zinc oxide nano rod at capillary surface, tube wall capillaceous is taken with scanning electron microscope, and its result as shown in Figure 1.
(2) with the glass capillary deionized water rinsing after step (1) processing, the ethanolic solution of 3-glycidoxypropyl trimethoxysilane of 3% (v/v) reinjects after the drying, soaking 2 hours, will be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall; With behind the alcohol flushing 110 ℃ of vacuum annealings 2 hours; Add the mouse-anti people HER2(ErbB-2 that contains 100 μ g/mL) monoclonal antibody PBS solution, soaked 5 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, and then adding concentration is the bovine serum albumin(BSA) about 10 mg/mL, soaked 1 hour, and got nano oxidized Zinc modified immune capillary.Its schematic diagram as shown in Figure 2, is fixed with zinc oxide nano rod at the kapillary inside surface as shown in Figure 2, is combined with mouse-anti people HER2 monoclonal antibody in the oxidisability nanometer rods.
Utilize the nano oxidized Zinc modified capillary detection tumor markers HER2 of preparation, concrete steps are as follows:
To contain 0 ng/mL respectively, 1 ng/mL, 5 ng/mL, 50 ng/mL, 500ng/mL and 1000 ng/mL tumor markers HER2(ErbB-2s, human epidermal growth factor receptor type 2) 10% human serum solution sucks respectively the kapillary device, arrest reaction 1 hour, again the anti-human HER2 polyclonal antibody of the goat of 5 μ g/mL is sucked kapillary 30 minutes after the discharge, then soaked kapillary 30 minutes with 5 μ g/mL CY3 mark goat IgG antibody.At last PBS solution and deionized water are flow through respectively kapillary, after the drying, read fluorescence signal with the fluorescent scanning instrument, then take fluorescence intensity as ordinate, the concentration of tumor markers HER2 is horizontal ordinate, the drawing standard curve, as shown in Figure 3.
Embodiment 2
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) getting concentration is that 1 mM newly prepares liquor potassic permanganate implantation glass kapillary, soaks 30 minutes activation glass capillaries; Then use deionized water rinsing, put into the 10 mM zinc nitrate solutions that contain 1% (v/v) monoethanolamine and 1% (v/v) ammoniacal liquor, growth is 60 minutes in 65 ℃ of water-baths, forms zinc oxide nano rod at capillary surface.
(2) with the glass capillary deionized water rinsing after step (1) processing, the ethanolic solution of 3-glycidoxypropyl trimethoxysilane of 1% (v/v) reinjects after the drying, soaked 1 hour, to be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall, with behind the alcohol flushing 110 ℃ of vacuum annealings after 3 hours, add and contain 50 μ g/mL mouse-anti people CEA(carcinomebryonic antigens, carcino-embryonic antigen) the PBS solution of monoclonal antibody, soaked 8 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, and then adding concentration is the bovine serum albumin(BSA) about 10 mg/mL, soaked 2 hours, and got nano oxidized Zinc modified immune capillary.
Utilize the nano oxidized Zinc modified capillary detection tumor markers CEA of preparation, concrete steps are as follows:
To contain 0ng/mL respectively, 1ng/mL, 5ng/mL, 50ng/mL, 500ng/mL, the 10% human serum solution of 1000 ng/mL tumor markers CEA adopt flow pumps to pump into respectively the kapillary device, be 5 μ L/min flowing reactives 1 hour at flow velocity, again the anti-human CEA polyclonal antibody of the goat of 1 μ g/mL was pumped into kapillary 30 minutes, then use the goat IgG antibody of 1 μ g/mL CY3 mark, the kapillary 30 minutes of flowing through.At last PBS solution and deionized water are flow through respectively kapillary, after the drying, read fluorescence signal with the fluorescent scanning instrument, then take fluorescence intensity as ordinate, the concentration of tumor markers CEA is horizontal ordinate, the drawing standard curve, as shown in Figure 4.
Embodiment 3
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) (preparation method is referring to Continuous Size Tuning of Monodisperse ZnO Colloidal Nanocrystal Clusters by a Microwave-Polyol Process and Their Application for Humidity Sensing to prepare the colloidal solution of zinc oxide nano-particle according to prior art, Xianluo Hu, Jingming Gong, Lizhi Zhang and Jimmy C. Yu. Adv. Mater. 2008,20, (24), 4845), again with colloidal solution implantation glass kapillary, soaked 20 minutes, then discharge colloidal solution, can repeat to soak, discharge step for several times, improve the adsorbance of zinc oxide nano-particle, then 150 ℃ of calcinings 60 minutes, the tube wall of immune capillary of preparation is taken with scanning electron microscope, and its result as shown in Figure 5.
(2) glass capillary after step (1) is processed injects the ethanolic solution of the 3-glycidoxypropyl trimethoxysilane of 1% (v/v), soaked 3 hours, to be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall, with behind the alcohol flushing 110 ℃ of vacuum annealings after 1 hour, add and contain 200 μ g/mL mouse-anti people PSA(prostate specific antigens, Prostate Specific Antigen) the PBS solution of monoclonal antibody, soaked 2 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, and then adding concentration is the bovine serum albumin(BSA) about 10 mg/mL, soaked 0.5 hour, and got nano oxidized Zinc modified immune capillary.Its schematic diagram as shown in Figure 6, is fixed with zinc oxide nano-particle at the kapillary inside surface as shown in Figure 6, is combined with mouse-anti people PSA monoclonal antibody at oxidisability nano particle and kapillary inside surface.
Utilize the nano oxidized Zinc modified capillary detection tumor markers PSA of preparation, concrete steps are as follows:
0ng/mL will be contained respectively, 1ng/mL, 10ng/mL, 100ng/mL, the 10% human serum solution of 1000 ng/mL tumor markers PSA sucks respectively the kapillary device, arrest reaction 0.5 hour, again the anti-human PSA polyclonal antibody of the goat of 10 μ g/mL is sucked kapillary 10 minutes after the discharge, then soaked kapillary 10 minutes with 10 μ g/mL CY3 mark goat IgG antibody, soak kapillary with PBS solution and deionized water at last, after the drying, read fluorescence signal with the fluorescent scanning instrument, then take fluorescence intensity as ordinate, tumor markers PSA concentration is horizontal ordinate, the drawing standard curve, as shown in Figure 7.
Embodiment 4
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) (preparation method is referring to Continuous Size Tuning of Monodisperse ZnO Colloidal Nanocrystal Clusters by a Microwave-Polyol Process and Their Application for Humidity Sensing to prepare the colloidal solution of zinc oxide nano-particle according to prior art, Xianluo Hu, Jingming Gong, Lizhi Zhang, and Jimmy C. Yu. Adv. Mater. 2008,20, (24), 4845), again with colloidal solution implantation glass kapillary, soaked 60 minutes, then discharge colloidal solution, can repeat to soak, discharge step for several times, improve the adsorbance of zinc oxide nano-particle, then 200 ℃ of calcinings 15 minutes.
(2) glass capillary after step (1) processing is injected the ethanolic solution that contains 5% (v/v) 3-glycidoxypropyl trimethoxysilane, soaked 1 hour, to be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall, with behind the alcohol flushing 110 ℃ of vacuum annealings after 3 hours, add and contain 100 μ g/mL mouse-anti people AFP(alpha-fetoproteins, alpha-fetal protein) the PBS solution of monoclonal antibody, soaked 5 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, and then adding concentration is the bovine serum albumin(BSA) about 10 mg/mL, soaked 0.5 hour, and got nano oxidized Zinc modified immune capillary.
Utilize the nano oxidized Zinc modified capillary detection tumor markers AFP of preparation, concrete steps are as follows:
0ng/mL will be contained respectively, 1 ng/mL, 10 ng/mL, the 10% human serum solution of 100 ng/mL and 1000 ng/mL tumor markers APF adopts respectively flow pumps with flow through take flow velocity as 500 μ L/min kapillary device 2 hours of sample solution, again the anti-human AFP polyclonal antibody of goat is flowed through kapillary 30 minutes, then use the goat IgG antibody of FITC mark, the kapillary 30 minutes of flowing through.At last PBS solution and deionized water are flow through respectively kapillary, after the drying, read fluorescence signal with the fluorescent scanning instrument, signal curve as shown in Figure 8.Then take fluorescence intensity as ordinate, tumor markers AFP concentration is horizontal ordinate, the drawing standard curve, as shown in Figure 9.
Embodiment 5
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) getting concentration is that 5 mM newly prepare 4 glass capillaries of liquor potassic permanganate injection, soaks 20 minutes activation glass capillaries; Then use deionized water rinsing, then put into the 50 mM zinc nitrate solutions that contain 5% (v/v) monoethanolamine and 5% (v/v) ammoniacal liquor, growth is 50 minutes in 80 ℃ of water-baths, forms zinc oxide nano rod at capillary surface.
(2) with the glass capillary deionized water rinsing after step (1) processing, the ethanolic solution of 3-glycidoxypropyl trimethoxysilane of 3% (v/v) reinjects after the drying, soaking 2 hours, will be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall; With behind the alcohol flushing 110 ℃ of vacuum annealings 2 hours; In different kapillaries, add mouse-anti people CEA, mouse-anti people PSA, mouse-anti people AFP and the 100ng/mL mouse-anti people HER2(contrast contain) the PBS solution of monoclonal antibody, soaked 5 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, then add again respectively concentration and be the bovine serum albumin(BSA) about 10 mg/mL, soaked 1 hour, and got nano oxidized Zinc modified immune capillary.
Utilize the nano oxidized Zinc modified kapillary of preparation to detect simultaneously the polycomponent tumor markers, concrete steps are as follows:
Four kinds of immune capillaries are adopted the flexible pipe series connection, adopt flow pumps with flow through take flow velocity as 300 μ L/min kapillary device 2 hours of sample solution the 10% human serum solution that contains respectively 100 ng/mL tumor markers CEA, PSA and APF, the PBS flow of solution that will contain again the anti-human CEA polyclonal antibody of goat, the anti-human PSA polyclonal antibody of goat and the anti-human AFP polyclonal antibody of goat was through kapillary 30 minutes, then use the goat IgG antibody of CY3 mark, the kapillary 30 minutes of flowing through.At last PBS solution and deionized water are flow through respectively kapillary, after the drying, read fluorescence signal with the fluorescent scanning instrument, schematic diagram and fluorescence signal curve are as shown in figure 10.
Embodiment 6
The preparation method of nano oxidized Zinc modified immune capillary, concrete steps are as follows:
(1) getting concentration is that 5 mM newly prepare 4 glass capillaries of liquor potassic permanganate injection, soaks 20 minutes activation glass capillaries; Then use deionized water rinsing, then put into the 50 mM zinc nitrate solutions that contain 5% (v/v) monoethanolamine and 5% (v/v) ammoniacal liquor, growth is 50 minutes in 80 ℃ of water-baths, forms zinc oxide nano rod at capillary surface.
(2) with the glass capillary deionized water rinsing after step (1) processing, the ethanolic solution of 3-glycidoxypropyl trimethoxysilane of 3% (v/v) reinjects after the drying, soaking 2 hours, will be to have formed silicon-oxygen covalent bond between 3-glycidoxypropyl trimethoxysilane and ZnO nano-structure and capillary tube inner wall; With behind the alcohol flushing 110 ℃ of vacuum annealings 2 hours; The PBS solution that in different kapillaries, adds respectively the mouse-anti people CEA, mouse-anti people PSA, mouse-anti people AFP and the 100ng/mL mouse-anti people HER2 monoclonal antibody (contrast) that contain 100 μ g/mL, soaked 5 hours, make epoxy bond and the reaction of the amino on the monoclonal antibody of 3-glycidoxypropyl trimethoxysilane form chemical bond, then add again respectively concentration and be the bovine serum albumin(BSA) about 10 mg/mL, soaked 1 hour, and got nano oxidized Zinc modified immune capillary.
Utilize the nano oxidized Zinc modified kapillary of preparation to detect simultaneously the polycomponent tumor markers, concrete steps are as follows:
Adopt flexible pipe in parallel four kinds of immune capillaries, as shown in figure 11, the 10% human serum solution that will contain tumor markers CEA, PSA and APF adopts respectively flow pumps with flow through take flow velocity as 300 μ L/min kapillary device 2 hours of sample solution, the PBS flow of solution that will contain again the anti-human CEA polyclonal antibody of goat, the anti-human PSA polyclonal antibody of goat and the anti-human AFP polyclonal antibody of goat was through kapillary 30 minutes, then use the goat IgG antibody of CY3 mark, the kapillary 30 minutes of flowing through.At last PBS solution and deionized water are flow through respectively kapillary, after the drying, read fluorescence signal with the fluorescent scanning instrument.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and not depart from the present invention that appended claims limits.
Claims (10)
1. nano oxidized Zinc modified immune capillary, it is characterized in that: then described immune capillary is formed in conjunction with detector probe on nano zine oxide by glass capillary inside surface growing nano zinc oxide.
2. described nano oxidized Zinc modified immune capillary according to claim 1, it is characterized in that: described nano zine oxide is zinc oxide nano rod or zinc oxide nano-particle.
3. described nano oxidized Zinc modified immune capillary according to claim 1, it is characterized in that: described detector probe is for detecting the monoclonal antibody of tumor markers.
4. each described nano oxidized Zinc modified immune capillary according to claim 1-3, it is characterized in that: described monoclonal antibody is anti-human epidermal growth factor acceptor 2 monoclonal antibody, anticancer embryonal antigen monoclonal antibody, anti-prostate-specific-antigen monoclonal antibody or anti-alpha-fetoprotein monoclonal antibody.
5. the preparation method of each described immune capillary of claim 1-4 is characterized in that, comprises the steps:
A. glass capillary is activated;
B. with the glass capillary inside surface growing nano zinc oxide of activation, get nano oxidized Zinc modified kapillary;
C. with the nano oxidized Zinc modified capillary surface of step b gained in conjunction with detector probe, get nano oxidized Zinc modified immune capillary.
6. the preparation method of described immune capillary according to claim 5 is characterized in that: described step a is to be that 1-10mM potassium permanganate soaked 15-30 minute with glass capillary concentration.
7. the preparation method of described immune capillary according to claim 5, it is characterized in that: described step b puts into the mixed solution that contains zinc nitrate, monoethanolamine and ammoniacal liquor with the activation glass capillary, growth is 30-60 minute in 65-95 ℃ of water-bath, the final concentration of zinc nitrate is 10-100mM in the described mixed solution, the final volume mark of monoethanolamine is 1-10%, and the final volume mark of ammoniacal liquor is 1-10%.
8. the preparation method of described immune capillary according to claim 5, it is characterized in that: described step c is with the nano oxidized Zinc modified kapillary deionized water rinsing of gained, inject after dry and contain the ethanolic solution that volume fraction is the 3-glycidoxypropyl trimethoxysilane of 1-5%, soaked 1-3 hour, with behind the alcohol flushing under 110 ℃ of conditions vacuum annealing 1-3 hour, the detector probe that adds again 50-300 μ g/mL was soaked 2-8 hour, soaked with bovine serum albumin(BSA) at last.
9. the preparation method of each described immune capillary according to claim 5-8, it is characterized in that: described step a and b replace with following steps: will soak 20-60 minute in the colloidal solution implantation glass kapillary of zinc oxide nano-particle, then under 150-200 ℃ of condition, calcined 15-60 minute, get nano oxidized Zinc modified kapillary.
10. each described nano oxidized Zinc modified immune capillary application in the preparation immunity testing equipment of claim 1-5.
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