CN104865299A - Preparation method and application of nanometer gold palladium/3aminopropyl triethoxy silanization manganic oxide PSA (prostate-specific antigen) sensor - Google Patents
Preparation method and application of nanometer gold palladium/3aminopropyl triethoxy silanization manganic oxide PSA (prostate-specific antigen) sensor Download PDFInfo
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- CN104865299A CN104865299A CN201510248437.1A CN201510248437A CN104865299A CN 104865299 A CN104865299 A CN 104865299A CN 201510248437 A CN201510248437 A CN 201510248437A CN 104865299 A CN104865299 A CN 104865299A
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Abstract
The invention relates to a preparation method and an application of a nanometer gold palladium/3aminopropyl triethoxy silanization manganic oxide PSA (prostate-specific antigen) sensor, and belongs to the technical fields of new functional materials and biosensing detections. According to the preparation method of the nanometer gold palladium/3aminopropyl triethoxy silanization manganic oxide PSA (prostate-specific antigen) sensor, a flower-like nanometer gold palladium and a manganic oxide composite material with good catalytic performances are used to prepare a sandwich type sensor, and therefore ultrasensitive detection on the prostate-specific antigen PSA is achieved, and a great significance of early diagnosis and judgements after cured on tumor markers is achieved.
Description
Technical field
The present invention relates to PSA transducer production method and the application of a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology structure.Specifically adopt flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide compound substance, prepare a kind of biology sensor detecting prostate specific antigen PSA, belong to new function material and bio-sensing detection technique field.
Background technology
Electrochemical immunosensor has that good, the sensitive height of selectivity, detectability are low, simple operation and other advantages, can be implemented in line Electrochemical Detection, be widely used at present the detection of tumor markers, therefore the present invention prepares a kind of detection realizing prostate specific antigen PSA based on flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology sensor.
At present because the d electron trajectory of noble metal does not fill up, surface is adsorption reaction thing easily, and moderate strength, be beneficial in the middle of being formed " reactive compound ", there is higher catalytic activity, there is the good characteristics such as anti-oxidant, corrosion-resistant simultaneously, become important catalyst material, be widely used in sensor field.Flower-like nanometer gold palladium has good biocompatibility and excellent catalytic activity, manganese sesquioxide managnic oxide also has good catalytic performance to hydrogen peroxide simultaneously, flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide compound substance is detected the label of antibody as prostate specific antigen PSA, the sensitivity of sensor can be improved, features such as reaching and detect the object of prostate specific antigen PSA, the method has that cost is low, simple to operate, high specificity, detection are quick.
Summary of the invention
An object of the present invention is the preparation based on flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology sensor.
Two of object of the present invention is the detections this electrochemical immunosensor being applied to prostate specific antigen PSA.
technical scheme of the present invention
1. the PSA transducer production method of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology structure
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water, then electrode is placed in 5 mmol/L potassium ferricyanide solutions, scans under-0.2 ~ 0.6 V current potential, make spike potential difference be less than 110 mV;
(2) take massfraction as the HPtCl of 1%, 2 mL
4for end liquid, scan 30s at voltage under-0.2V, obtain the electrode of electrodeposition Pt nano particle;
(3) the prostate specific antigen PSA capture antibody Ab of 6 μ L, 5 ~ 10 μ g/mL is dripped
1, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 ~ 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the prostate specific antigen PSA solution of 0.0005 ~ 20 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) continue dropping 4 ~ 6 μ L and detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, the PSA sensor that obtained a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds.
2. detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2the preparation of solution
(1) preparation of 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide
0.5 ~ 2.0 g glucose is joined the Mn (NO of 30 ~ 80 mL, 0.60 mol/L
3)
2in solution, be moved in autoclave after 10 min, react 18 h, be cooled to room temperature at 180 DEG C, centrifugal, with ethanol and milli-Q water, obtain presoma after 80 DEG C of dryings, high-temperature calcination 3 h obtains manganese sesquioxide managnic oxide at 550 DEG C subsequently;
Get 0.05 ~ 1.5 g manganese sesquioxide managnic oxide and be placed in there-necked flask, add 3-aminopropyl triethoxysilane and the 10 mL absolute ethyl alcohols of 0.1 ~ 3 mL, be heated to 80 DEG C and keep 5 ~ 10 h, cool to room temperature, gained potpourri through washing, centrifuging, vacuum drying at 45 DEG C, i.e. obtained 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(2) preparation of flower-like nanometer gold palladium
1 mL is contained 2 ~ 8 mmol/L HAuCl
4with 2 ~ 8 mmol/L K
2pdCl
4aqueous solution be added in 47 mL ultrapure waters, after adding 0.5 ~ 2 mL, 0.1 mol/L sodium citrate 15 s, under agitation, 0.5 ~ 2 mL, 5 mg/mL PVP are dropwise added, stir 30 min, be re-dispersed in ultrapure water after potpourri is centrifugal, washing, the solution of obtained flower-like nanometer gold palladium;
(3) preparation of antibody labeling thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is detected
Respectively the 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide of 10 ~ 30 mg is distributed in the solution of flower-like nanometer gold palladium of 5 ~ 100 mL, under room temperature, shakes 12 h; Potpourri through washing, centrifuging, vacuum drying at 35 DEG C, obtained flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(4) antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab is detected
2the preparation of solution
The flower-like nanometer gold of 1 ~ 3 mg palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is scattered in 1 mL ultrapure water, add the detection antibody-solutions of 1 mL, 5 ~ 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, saves backup at 4 DEG C.
3. the detection method of prostate specific antigen PSA
(1) three-electrode system is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.0, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) after background current tends towards stability, in the phosphate buffered solution of 10 mL, pH=7.0, the hydrogen peroxide solution of 10 μ L, 5 mol/L is injected every 50 s, record current changes, linear according to the concentration of gained current differential and prostate specific antigen PSA, drawing curve;
(3) detection of prostate specific antigen PSA in sample is carried out in the drafting of foundation working curve, and the result of detection can checking at working curve.
The results show, time of the present invention, the current differential of m-electric current and the concentration of prostate specific antigen PSA keep good linear relationship within the scope of 0.0005 ~ 20 ng/mL, and detectability reaches 0.11 pg/mL.
useful achievement of the present invention
(1) the Pt nano particle electroplated has good electric conductivity, and good biocompatibility and stability, and in addition, it in conjunction with a large amount of capture antibodies, can improve the charge capacity of capture antibody on its surface, add transducer sensitivity and stability.
(2) flower-like nanometer gold palladium and 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide all have good catalytic activity, it can be used as the label detecting antibody, can reach the effect that signal amplifies, thus improve the sensitivity of biology sensor.
(3) electrochemical immunosensor prepared of the present invention is for the detection of prostate specific antigen PSA, and the response time is short, and detectability is low, and the range of linearity is wide, can realize simple, quick, highly sensitive and specific detection.
Embodiment
embodiment 1the PSA transducer production method that a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water, then electrode is placed in 5 mmol/L potassium ferricyanide solutions, scans under-0.2 ~ 0.6 V current potential, make spike potential difference be less than 110 mV;
(2) take massfraction as the HPtCl of 1%, 2 mL
4for end liquid, scan 30s at voltage under-0.2V, obtain the electrode of electrodeposition Pt nano particle;
(3) the prostate specific antigen PSA capture antibody Ab of 6 μ L, 5 μ g/mL is dripped
1, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the prostate specific antigen PSA solution of 0.0005 ~ 20 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) continue dropping 4 μ L and detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, the PSA sensor that obtained a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds.
embodiment 2the PSA transducer production method that a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water, then electrode is placed in 5 mmol/L potassium ferricyanide solutions, scans under-0.2 ~ 0.6 V current potential, make spike potential difference be less than 110 mV;
(2) take massfraction as the HPtCl of 1%, 2 mL
4for end liquid, scan 30s at voltage under-0.2V, obtain the electrode of electrodeposition Pt nano particle;
(3) the prostate specific antigen PSA capture antibody Ab of 6 μ L, 7 μ g/mL is dripped
1, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 10 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the prostate specific antigen PSA solution of 0.0005 ~ 20 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) continue dropping 5 μ L and detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, the PSA sensor that obtained a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds.
embodiment 3the PSA transducer production method that a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water, then electrode is placed in 5 mmol/L potassium ferricyanide solutions, scans under-0.2 ~ 0.6 V current potential, make spike potential difference be less than 110 mV;
(2) take massfraction as the HPtCl of 1%, 2 mL
4for end liquid, scan 30s at voltage under-0.2V, obtain the electrode of electrodeposition Pt nano particle;
(3) the prostate specific antigen PSA capture antibody Ab of 6 μ L, 10 μ g/mL is dripped
1, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the prostate specific antigen PSA solution of 0.0005 ~ 20 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) continue dropping 6 μ L and detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, the PSA sensor that obtained a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds.
embodiment 4detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2the preparation of solution
(1) preparation of 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide
0.5 g glucose is joined the Mn (NO of 30 mL, 0.60 mol/L
3)
2in solution, be moved in autoclave after 10 min, react 18 h, be cooled to room temperature at 180 DEG C, centrifugal, with ethanol and milli-Q water, obtain presoma after 80 DEG C of dryings, high-temperature calcination 3 h obtains manganese sesquioxide managnic oxide at 550 DEG C subsequently;
Get 0.05 g manganese sesquioxide managnic oxide and be placed in there-necked flask, add 3-aminopropyl triethoxysilane and the 10 mL absolute ethyl alcohols of 0.1 mL, be heated to 80 DEG C and keep 5 h, cool to room temperature, gained potpourri through washing, centrifuging, vacuum drying at 45 DEG C, i.e. obtained 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(2) preparation of flower-like nanometer gold palladium
1 mL is contained 2 mM HAuCl
4with 2 mmol/L K
2pdCl
4aqueous solution be added in 47 mL ultrapure waters, after adding 0.5 mL, 0.1 mol/L sodium citrate 15 s, under agitation, 0.5 mL, 5 mg/mL PVP are dropwise added, stir 30 min, be re-dispersed in ultrapure water after potpourri is centrifugal, washing, the solution of obtained flower-like nanometer gold palladium;
(3) preparation of antibody labeling thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is detected
Respectively the 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide of 10 mg is distributed in the solution of flower-like nanometer gold palladium of 50 mL, under room temperature, shakes 12 h; Potpourri through washing, centrifuging, vacuum drying at 35 DEG C, obtained flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(4) antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab is detected
2the preparation of solution
The flower-like nanometer gold of 1 mg palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is scattered in 1 mL ultrapure water, add the detection antibody-solutions of 1 mL, 5 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, saves backup at 4 DEG C.
embodiment 5detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2the preparation of solution
(1) preparation of 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide
1.0 g glucose are joined the Mn (NO of 50 mL, 0.60 mol/L
3)
2in solution, be moved in autoclave after 10 min, react 18 h, be cooled to room temperature at 180 DEG C, centrifugal, with ethanol and milli-Q water, obtain presoma after 80 DEG C of dryings, high-temperature calcination 3 h obtains manganese sesquioxide managnic oxide at 550 DEG C subsequently;
Get 1.0 g manganese sesquioxide managnic oxides and be placed in there-necked flask, add 3-aminopropyl triethoxysilane and the 10 mL absolute ethyl alcohols of 1 mL, be heated to 80 DEG C and keep 8 h, cool to room temperature, gained potpourri through washing, centrifuging, vacuum drying at 45 DEG C, i.e. obtained 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(2) preparation of flower-like nanometer gold palladium
1 mL is contained 3 mmol/L HAuCl
4with 5 mmol/L K
2pdCl
4aqueous solution be added in 47 mL ultrapure waters, after adding 1 mL, 0.1 mol/L sodium citrate 15 s, under agitation, 1 mL, 5 mg/mL PVP are dropwise added, stir 30 min, be re-dispersed in ultrapure water after potpourri is centrifugal, washing, the solution of obtained flower-like nanometer gold palladium;
(3) preparation of antibody labeling thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is detected
Respectively the 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide of 20 mg is distributed in the solution of flower-like nanometer gold palladium of 80 mL, under room temperature, shakes 12 h; Potpourri through washing, centrifuging, vacuum drying at 35 DEG C, obtained flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(4) antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab is detected
2the preparation of solution
The flower-like nanometer gold of 2 mg palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is scattered in 1 mL ultrapure water, add the detection antibody-solutions of 1 mL, 10 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, saves backup at 4 DEG C.
embodiment 6detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2the preparation of solution
(1) preparation of 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide
2.0 g glucose are joined the Mn (NO of 80 mL, 0.60 mol/L
3)
2in solution, be moved in autoclave after 10 min, react 18 h, be cooled to room temperature at 180 DEG C, centrifugal, with ethanol and milli-Q water, obtain presoma after 80 DEG C of dryings, high-temperature calcination 3 h obtains manganese sesquioxide managnic oxide at 550 DEG C subsequently;
Get 1.5 g manganese sesquioxide managnic oxides and be placed in there-necked flask, add 3-aminopropyl triethoxysilane and the 10 mL absolute ethyl alcohols of 3 mL, be heated to 80 DEG C and keep 10 h, cool to room temperature, gained potpourri through washing, centrifuging, vacuum drying at 45 DEG C, i.e. obtained 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(2) preparation of flower-like nanometer gold palladium
1 mL is contained 8 mmol/L HAuCl
4with 8 mmol/L K
2pdCl
4aqueous solution be added in 47 mL ultrapure waters, after adding 2 mL, 0.1 mol/L sodium citrate 15 s, under agitation, 2 mL, 5 mg/mL PVP are dropwise added, stir 30 min, be re-dispersed in ultrapure water after potpourri is centrifugal, washing, the solution of obtained flower-like nanometer gold palladium;
(3) preparation of antibody labeling thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is detected
Respectively the 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide of 30 mg is distributed in the solution of flower-like nanometer gold palladium of 100 mL, under room temperature, shakes 12 h; Potpourri through washing, centrifuging, vacuum drying at 35 DEG C, obtained flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(4) antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab is detected
2the preparation of solution
The flower-like nanometer gold of 3 mg palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is scattered in 1 mL ultrapure water, add the detection antibody-solutions of 1 mL, 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, saves backup at 4 DEG C.
embodiment 7the detection of prostate specific antigen PSA
(1) three-electrode system is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.0, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) after background current tends towards stability, in the phosphate buffered solution of 10 mL, pH=7.0, the hydrogen peroxide solution of 10 μ L, 5 mol/L is injected every 50 s, record current changes, linear according to the concentration of gained current differential and prostate specific antigen PSA, drawing curve;
(3) carry out the detection of prostate specific antigen PSA in sample according to the drafting of working curve, the result of detection can checking at working curve;
The results show, time of the present invention, the current differential of m-electric current and the concentration of prostate specific antigen PSA keep good linear relationship within the scope of 0.0005 ~ 20 ng/mL, and detectability reaches 0.11 pg/mL.
Claims (3)
1. the PSA transducer production method that flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds, is characterized in that, comprise the following steps:
(1) use the alundum (Al2O3) burnishing powder of 1.0,0.3,0.05 μm to glass-carbon electrode polishing successively, clean up with ultrapure water, then electrode is placed in 5 mmol/L potassium ferricyanide solutions, scans under-0.2 ~ 0.6 V current potential, make spike potential difference be less than 110 mV;
(2) take massfraction as the HPtCl of 1%, 2 mL
4for end liquid, scan 30s at voltage under-0.2V, obtain the electrode of electrodeposition Pt nano particle;
(3) the prostate specific antigen PSA capture antibody Ab of 6 μ L, 5 ~ 10 μ g/mL is dripped
1, dry at 4 DEG C, ultrapure water;
(4) drip 3 μ L, massfraction be the bovine serum albumin solution of 5 ~ 15 mg/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(5) drip 6 μ L concentration be the prostate specific antigen PSA solution of 0.0005 ~ 20 ng/mL to electrode surface, dry at 4 DEG C, ultrapure water;
(6) continue dropping 4 ~ 6 μ L and detect antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, to electrode surface, is placed in 4 DEG C of refrigerators and hatches 1 h, after cleaning, dry, the PSA sensor that obtained a kind of flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds.
2. the PSA transducer production method that a kind of flower-like nanometer gold palladium as claimed in claim 1/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide biology builds, described detection antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2the preparation of solution, is characterized in that, comprises the following steps:
(1) preparation of 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide
0.5 ~ 2.0 g glucose is joined the Mn (NO of 30 ~ 80 mL, 0.60 mol/L
3)
2in solution, be moved in autoclave after 10 min, react 18 h, be cooled to room temperature at 180 DEG C, centrifugal, with ethanol and milli-Q water, obtain presoma after 80 DEG C of dryings, high-temperature calcination 3 h obtains manganese sesquioxide managnic oxide at 550 DEG C subsequently;
Get 0.05 ~ 1.5 g manganese sesquioxide managnic oxide and be placed in there-necked flask, add 3-aminopropyl triethoxysilane and the 10 mL absolute ethyl alcohols of 0.1 ~ 3 mL, be heated to 80 DEG C and keep 5 ~ 10 h, cool to room temperature, gained potpourri through washing, centrifuging, vacuum drying at 45 DEG C, i.e. obtained 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(2) preparation of flower-like nanometer gold palladium
1 mL is contained 2 ~ 8 mmol/L HAuCl
4with 2 ~ 8 mmol/L K
2pdCl
4aqueous solution be added in 47 mL ultrapure waters, after adding 0.5 ~ 2 mL, 0.1 mol/L sodium citrate 15 s, under agitation, 0.5 ~ 2 mL, 5 mg/mL PVP are dropwise added, stir 30 min, be re-dispersed in ultrapure water after potpourri is centrifugal, washing, the solution of obtained flower-like nanometer gold palladium;
(3) preparation of antibody labeling thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is detected
Respectively the 3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide of 10 ~ 30 mg is distributed in the solution of flower-like nanometer gold palladium of 5 ~ 100 mL, under room temperature, shakes 12 h; Potpourri through washing, centrifuging, vacuum drying at 35 DEG C, obtained flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide;
(4) antibody hatching thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab is detected
2the preparation of solution
The flower-like nanometer gold of 1 ~ 3 mg palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide is scattered in 1 mL ultrapure water, add the detection antibody-solutions of 1 mL, 5 ~ 12 μ g/mL, vibrate hatching 12 h at 4 DEG C, centrifuging, lower sediment be scattered in the pH=7.4 phosphate buffered solution of 1 mL, 1/15 mol/L, the obtained antibody that detects hatches thing-flower-like nanometer gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide-Ab
2solution, saves backup at 4 DEG C.
3. the immunosensor prepared of preparation method as claimed in claim 1 is for the detection method of prostate specific antigen PSA, it is characterized in that, comprises following analytical procedure:
(1) three-electrode system is adopted to measure, using the immunosensor prepared by claim 1 as working electrode, saturated calomel electrode is to electrode, platinum electrode is auxiliary electrode, in the phosphate buffered solution of pH=7.0, during employing, the method for m-electric current scans, and input voltage is-0.4 V, working time 400 s;
(2) after background current tends towards stability, in the phosphate buffered solution of 10 mL, pH=7.0, the hydrogen peroxide solution of 10 μ L, 5 mol/L is injected every 50 s, record current changes, linear according to the concentration of gained current differential and prostate specific antigen PSA, drawing curve;
(3) detection of prostate specific antigen PSA in sample is carried out in the drafting of foundation working curve, and the result of detection can checking at working curve.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510248437.1A CN104865299B (en) | 2015-05-16 | 2015-05-16 | The PSA transducer production method of a kind of nm of gold palladium/3-aminopropyl triethoxysilane manganese sesquioxide managnic oxide and application |
Applications Claiming Priority (1)
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| CN105699464A (en) * | 2016-03-05 | 2016-06-22 | 济南大学 | Preparation method and application of immunity sensor based on cuprous oxide doped palladium nanoparticle marks |
| CN106556700A (en) * | 2016-11-01 | 2017-04-05 | 济南大学 | Electrochemiluminescimmunosensor immunosensor and its application are built based on C60 self characters |
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| CN106556700A (en) * | 2016-11-01 | 2017-04-05 | 济南大学 | Electrochemiluminescimmunosensor immunosensor and its application are built based on C60 self characters |
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| CN104865299B (en) | 2016-04-20 |
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