CN106290878A - The preparation method of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot - Google Patents

The preparation method of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot Download PDF

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CN106290878A
CN106290878A CN201610815157.9A CN201610815157A CN106290878A CN 106290878 A CN106290878 A CN 106290878A CN 201610815157 A CN201610815157 A CN 201610815157A CN 106290878 A CN106290878 A CN 106290878A
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常津
武玉东
宫晓群
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Tianjin University
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    • G—PHYSICS
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label

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Abstract

The present invention relates to the preparation method of a kind of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot；By EDC (1 ethyl (3 dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate) activation, the carboxyl of quantum dot and pancreatic tumour mark CA242 antibody A b2Amino generate firm chemical bond, probe, pancreatic tumour mark CA242 and be embedded in another kind of pancreatic tumour mark CA242 antibody A b of test strips1By the effect between antibody antigen, form a kind of similar sandwich structure, the detection pancreatic tumour mark CA242 of qualitative, quantitative.Preparation process is simple, is suitable for industrialization and produces；According to the fluorescent characteristic of quantum dot, can qualitative and quantitative detection pancreatic tumour mark CA242, and specificity is good；Whole detection process, with low cost and operation the easiest, establish a kind of pancreatic tumour mark CA242 detection new method.

Description

The system of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot Preparation Method
Technical field
The present invention relates to disease detection technical field, more particularly to a kind of pancreatic tumour mark based on quantum dot The preparation method of will thing CA242 immuno-chromatographic test paper strip.
Background technology
Tumor markers (Tumor Marker) is the chemicals that reflection tumor exists.They or be not present in normal Adult organizes and is detected in embryonal tissue, or the content in tumor tissues substantially exceeds the content in normal structure, they Existence or quantitative change can point out the character of tumor, so as to understanding the tissue of tumor and occurring, cell differentiation, cell function, with side The diagnosis of tumor, classification, Index for diagnosis and treatment is helped to instruct.CA242 is a kind of sialylated sugar antigen, can be by colon One of a series of monoclonal antibodies that JEG-3 obtains through hybridoma technology CA242 is identified, it is that one is present in many devices In official's malignant tumor, the glycoprotein in mucin types is CanAg, i.e. can not be with LewisA type antigen-reactive, can not be with saliva The galactoside reaction of acidifying.Immunochemistry research has shown that it is different from tumor-associated mucin known to other such as: In CA199, CA50, CA125, CA153 etc., Healthy People and benign disease serum, content is relatively low.CA242 is to be applied in recent years face A kind of tumor markers that bed is newer, cancer of pancreas and the preferable tumor markers of colon cancer.CA242 is a kind of sialylated sheath Sugar lipid antigen, the most always expresses together with CA50, but both is by different monoclonal antibody identification.The most all by with In malignant tumor of digestive tract especially cancer of pancreas, the diagnosis of colorectal cancer, compared with CA19-9, CA50, the CA242 of a new generation exists Sensitivity, specificity in cancer of pancreas, carcinoma of gallbladder and digestive tract cancer are higher, and (CA50, CA19-9 are easily strongly fragrant by liver function and bile Long-pending impact, damages note disease in optimum obstructive jaundice and liver parenchyma and false positive often occurs).CA242 is a kind of viscous Protein type carbohydrate antigen, can be as the preferable tumor markers of cancer of pancreas and colon cancer, and its sensitivity is similar with CA19-9, but special Property, diagnosis efficiency are then better than CA19-9.CA242 normal value < 20 (unit: IU/ml)
In semi-conducting material, tiny crystals is commonly referred to as quantum dot (quantum dot).This quantum dot can be electricity Son is locked in a three dimensions the most small, and when having light beam to irradiate up when, electrons is excited and jumps to Higher energy level.When these electronics return to the most relatively low energy level when, the light beam that wavelength is certain can be launched.Quantum now Point is applied at biological experiment indoor in large quantities, helps research worker to determine structure or the activity of biological cell.Work as quantum dot Can be produced various color when of optical pulse irradiation, the most senior optical microscope is just it is observed that this coloured silk Coloured light.If quantum dot being attached to need on the object of research, research worker just it will be seen that the activity of material.The most such as This, quantum dot can also be used to follow the trail of medicine activity in vivo or study cell and the structure of tissue in the patient.Quantum Point can produce the light of multiple color, and the color of light depends on the size of quantum dot.Research worker has produced and can produce More than the quantum dot of 12 kinds of color fluorescence, and more color can be produced theoretically.So, swashing when certain wavelength Light carries out irradiation when excite to multiple quantum point, can simultaneously observe multiple color, carry out multiple measurement simultaneously.Biological Quantum dot used in research needs covering last layer material so that specific biomolecule can be followed the trail of, and can apply doctor Learn in imaging technique.External scientist has applied quantum dot marked tumor cell to carry out being correlated with by living imaging system Study current quantum dot and be widely used in the aspects such as spike, imaging and labelling because having excellent optical property.And Immuno-chromatographic test paper strip detection then because it the rapidest, cheap, can wait advantage whenever and wherever possible at detection field In special consequence.So intending herein developing pancreatic tumour mark CA242 immune chromatography test paper based on quantum dot The preparation method of bar, detects carbohydrate antigen CA242.
Summary of the invention
In view of the optical property that tumor markers critical role in lesion detection, this nanoparticle of quantum dot are unique And chromatographic technique is easy and advantage in price.We are by incorporating quantum point and two kinds of technology of immuno-chromatographic test paper strip, profit React with the amino of quantum dot carboxyl and tumor markers corresponding antibodies, prepare quantum dot fluorescence probe.Probe, tumor markers With the another kind of tumor markers antibody being embedded in test strips passes through the effect between antibody antigen, formed a kind of similar sandwich Structure.To carbohydrate antigen CA242, this sialylated glycosyl sphingolipid class antigen carries out the detection of qualitative, quantitative, sets up tumor mark The new method of will analyte detection, is devoted to the rapidest, cheap, qualitative, quantitative, early diagnosis of tumor method easy and simple to handle.
Technical scheme is as follows:
A kind of preparation method of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot；Its step As follows:
1) by quantum dot (Quantum dots, QDs), 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimine hydrochloric acid Salt (EDC) and PBS add reactor, are rotating the carboxyl activating quantum dot on hybrid frame, are centrifuged and obtain immunity quantum Point immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2Mix with above-mentioned immune QDs, add PBS buffering Liquid, is rotating coupled antibody 2～3h on hybrid frame, and the centrifugal quantum dot-pancreatic tumour mark traget antibody fluorescence that obtains is visited Pin QDs-Ab2, then close QDs-Ab with bovine serum albumin (Albumin from bovine serum, BSA)2Fluorescent probe Upper unreacted carboxyl, processes sample pad, pad treatment fluid process pad with sample pad treatment fluid simultaneously and dries, then Process pad with above-mentioned fluorescent probe solution and dry；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2～3mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Described quantum dot QDs: pancreatic tumour mark CA242 traget antibody Ab2Mass fraction ratio=1:10～50.
Described quantum dot QDs:1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate EDC mass fraction ratio For 1:4000～10000.
Described PBS is the PBS of 0.01M pH 7.4, in order to keep pancreatic tumour mark CA242 The immunocompetence of antibody.
Described pad treatment fluid is the sucrose containing mass volume ratio 5%, the bovine serum albumin of mass volume ratio 5% BSA, the Polyethylene Glycol PEG of mass volume ratio 3%, the polyoxyethylene sorbitan monolaurate Tween-20 of volume ratio 2% PBS solution；Sample pad treatment fluid is the PBS solution of the Tween-20 of volume ratio 0.5%.
Dripping the pancreatic tumour mark CA242 of variable concentrations, Criterion curve in sample pad, qualitative, quantitative is examined Survey pancreatic tumour mark CA242.(wherein 0ng/mL correspondence hyclone FBS makees variable concentrations preferably 0～1000ng/mL For negative control).
By the activation of EDC (1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate), quantum dot Carboxyl and pancreatic tumour mark CA242 traget antibody Ab2The amino of (molecular weight 150000～200000) generates firm Chemical bond, is dispersed in after centrifugal purification in the PBS containing bovine serum albumin, obtains QDs-Ab2Fluorescent probe.By pancreas Adenocarcinoma tumor mark CA242 traget antibody Ab1Uniformly it is sprayed on nitrocellulose filter as detection line T, and by goat-anti mouse-anti Body Ab3As nature controlling line C on nitrocellulose filter.By the four of test strips sample segment pads, pad, adsorptive pads and nitric acid Cellulose membrane assembles together, finally obtains pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.Dropping detection Sample is in sample pad, and detection sample can move forward under the chromatography effect of adsorptive pads: if detection sample has pancreas cancerous protuberance Tumor markers CA242 exists, probe QDs-Ab2, pancreatic tumour mark CA242 and the pancreas that is embedded on nitrocellulose filter Adenocarcinoma tumor mark CA242 coated antibody Ab1By the effect between antibody antigen, T line forms one similar sandwich Structure QDs-Ab2-CA242-Ab1, and on C line, form QDs-Ab2-Ab3Structure, now T line and C line are the brightest；If Detection sample does not has pancreatic tumour mark CA242 antigen, then probe QDs-Ab2Only and be embedded in celluloid Sheep anti-mouse antibody on film, forms QDs-Ab2-Ab3Structure, now C line is bright and T line does not works.
Novel based on quantum dot immuno-chromatographic test paper strip advantage prepared by the present invention is:
1. use the nano material of this optical property with excellence of quantum dot as fluorescence probe light source, quantum dot table Face can combine with the amino groups of protein molecular with a large amount of carboxyls, be widely used in showing through modification The aspects such as track, imaging and labelling, are applied to lesion detection field by nanotechnology.
2. use immunochromatography technique this rapidest, cheap, the conventional art of advantage can be waited whenever and wherever possible, Contribute to realizing POC detection.
3. use the firm chemical bond that the reaction between carboxyl and the amino of protein molecular of quantum dot is formed, will significantly Improve the ability of test strips opposing non-specific adsorption, quantum dot fluorescence intensity and pancreatic tumour mark CA242 concentration it Between relation, Criterion curve, qualitative and quantitative detection can be realized simultaneously.
As it is shown in figure 1, quantum used by the pancreatic tumour mark CA242 test strips based on quantum dot prepared of the present invention Point dispersibility is homogeneous, and particle diameter is less and a carrier is surrounded by multiple quantum dot；As in figure 2 it is shown, the present invention prepare based on quantum The standard curve of the pancreatic tumour mark CA242 test strips of point is y=-5E-06x2+0.0042x+0.2016R2= 0.9901；As it is shown on figure 3, pancreatic tumour mark CA242 test strips based on quantum dot prepared by the present invention is non-specific Adsorb relatively low；As shown in Figure 4, Figure 5, the pancreatic tumour mark CA242 test strips based on quantum dot that prepared by the present invention is The good response time is 25min, and optimal injection volume is 40uL.
Accompanying drawing explanation
The quantum dot transmission electricity of pancreatic tumour mark CA242 test strips based on quantum dot prepared by Fig. 1 present invention Mirror photo.
The standard curve of pancreatic tumour mark CA242 test strips based on quantum dot prepared by Fig. 2 present invention.
The specificity experiments of pancreatic tumour mark CA242 test strips based on quantum dot prepared by Fig. 3 present invention.
The response time of pancreatic tumour mark CA242 test strips based on quantum dot prepared by Fig. 4 present invention optimizes Experiment.
The injection volume optimization of pancreatic tumour mark CA242 test strips based on quantum dot prepared by Fig. 5 present invention Experiment.
Detailed description of the invention
In following case study on implementation, the invention will be further elaborated, but the invention is not restricted to this.
Case study on implementation 1:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 2h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 2:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:30), add PBS, rotating coupled antibody 2h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 3:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:50), add PBS, rotating coupled antibody 2h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 4:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:6000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 2.5h on hybrid frame, be centrifuged and obtain quantum dot-pancreas Tumor mark CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from Bovine serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample with sample pad treatment fluid simultaneously Pad, pad treatment fluid process pad and dry, and then process pad with above-mentioned fluorescent probe solution and dry；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2.5mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 5:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:10000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 2.5h on hybrid frame, be centrifuged and obtain quantum dot-pancreas Tumor mark CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from Bovine serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample with sample pad treatment fluid simultaneously Pad, pad treatment fluid process pad and dry, and then process pad with above-mentioned fluorescent probe solution and dry；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 6:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 3h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 3mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Case study on implementation 7:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 2h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
Sample pad dropping concentration be 1000,500,250,100,50, the pancreatic tumour mark CA242 of 10IU/mL, Matching draws standard curve；Sample pad drip respectively PSA antigen, AFP antigen, CEA antigen, CA125 antigen, CA153 antigen, CA199 antigen, FCS and HCG, draw non-specific curve.
Case study on implementation 7:
1) by quantum dot (Quantum dots, QDs) that mass fraction proportioning is 1:4000 and 1-ethyl-(3-dimethyl Aminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC) addition reactor, add PBS, activate quantum on hybrid frame rotating The carboxyl of point, is centrifuged and obtains immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2(QDs and Ab is mixed with above-mentioned immune QDs2Quality Mark proportioning is 1:10), add PBS, rotating coupled antibody 2h on hybrid frame, be centrifuged and obtain quantum dot-cancer of pancreas Tumor markers CA242 traget antibody fluorescent probe QDs-Ab2, then with bovine serum albumin (Albumin from bovine Serum, BSA) close QDs-Ab2Unreacted carboxyl on fluorescent probe, processes sample pad, combination with sample pad treatment fluid simultaneously Pad treatment fluid processes pad and dries, and then processes pad with above-mentioned fluorescent probe solution and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking Add 2mm, obtain pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
After sample pad dropping antigen 5,10,15,20,25,30min respectively test strip fluorescent value, choose optimum response Time.Sample pad drips the consumption of pancreatic tumour mark CA242 respectively and is respectively 30,40,50,75,100,150, chooses Optimal injection volume.

Claims (7)

1. the preparation method of a pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot；It is characterized in that Step is as follows:
1) by quantum dot (Quantum dots, QDs), 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate And PBS adds reactor (EDC), rotate the carboxyl activating quantum dot on hybrid frame, centrifugal obtain immunity quantum dot immune QDs；
2) by pancreatic tumour mark CA242 traget antibody Ab2Mix with above-mentioned immune QDs, add PBS, Rotate coupled antibody 2～3h on hybrid frame, be centrifuged and obtain quantum dot-pancreatic tumour mark traget antibody fluorescent probe QDs- Ab2, then close QDs-Ab with bovine serum albumin (Albumin from bovine serum, BSA)2On fluorescent probe the most anti- The carboxyl answered, processes sample pad, pad treatment fluid process pad with sample pad treatment fluid simultaneously and dries, then with above-mentioned Fluorescent probe solution processes pad and dries；
3) prepared by test strips: adsorptive pads, nitrocellulose filter, sample pad and pad are fitted together, every stacking add 2～ 3mm, obtains pancreatic tumour mark CA242 quantum dot immune chromatograph test strip.
2. the method for claim 1, is characterized in that quantum dot QDs:1-ethyl-(3-dimethylaminopropyl) phosphinylidyne Diimmonium salt hydrochlorate EDC mass fraction is than for 1:4000～10000.
3. the method for claim 1, is characterized in that quantum dot QDs: pancreatic tumour mark traget antibody Ab2Quality Score ratio=1:10～50.
4. the method for claim 1, is characterized in that the PBS that PBS used is 0.01M pH 7.4, uses To keep the immunocompetence of pancreatic tumour mark antibody.
5. the method for claim 1, is characterized in that pad treatment fluid is the sucrose containing mass volume ratio 5%, matter Amount the bovine serum albumin BSA of volume ratio 5%, the Polyethylene Glycol PEG of mass volume ratio 3%, the polyoxyethylene mountain of volume ratio 2% The PBS solution of pears alcohol monolaurate Tween-20；Sample pad treatment fluid is the PBS solution of the Tween-20 of volume ratio 0.5%.
6. the method for claim 1, it is characterized in that according to test strips sandwich structure capture quantum dot fluorescence intensity and Relation between normal pancreatic tumor mark CA242 concentration, Criterion curve, qualitative and quantitative detection pancreatic tumour mark Will thing CA242.
7. method as claimed in claim 6, is characterized in that dripping the pancreatic tumour mark of variable concentrations in sample pad CA242, variable concentrations is 0～1000mIU/mL, Criterion curve.
CN201610815157.9A 2016-09-10 2016-09-10 The preparation method of pancreatic tumour mark CA242 immuno-chromatographic test paper strip based on quantum dot Pending CN106290878A (en)

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CN104991063A (en) * 2015-06-25 2015-10-21 天津大学 Method for preparing carcino-embryonic antigen immunochromatography test strip based on quantum dots
CN105044339A (en) * 2015-06-25 2015-11-11 天津大学 Novel breast cancer quantum dot immunochromatographic test strip preparation method
CN104991068A (en) * 2015-07-02 2015-10-21 天津大学 Preparation method of AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots)
CN105044340A (en) * 2015-07-02 2015-11-11 天津大学 Preparation method of quantum dot based prostate specific antigen immuno-chromatographic test strip

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