CN108535240A - The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence - Google Patents
The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence Download PDFInfo
- Publication number
- CN108535240A CN108535240A CN201810331532.1A CN201810331532A CN108535240A CN 108535240 A CN108535240 A CN 108535240A CN 201810331532 A CN201810331532 A CN 201810331532A CN 108535240 A CN108535240 A CN 108535240A
- Authority
- CN
- China
- Prior art keywords
- bovine serum
- serum albumin
- trypsase
- luminol
- chemiluminescence
- 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.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/976—Trypsin; Chymotrypsin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of method detecting trypsase with bovine serum albumin copper nano-cluster catalysis luminol chemiluminescence, which specifically includes：S1：Take trypsin solution to be measured, bovine serum albumin copper nanocluster catalyst, Constant temperature hatch after mixing；S2：Luminol hydrogen peroxide detection liquid progress chemiluminescence reaction is added into mixed liquor, the concentration of trypsase to be measured is calculated according to its chemiluminescence intensity and the standard curve of foundation.The present invention detection method, it is cheap, detection speed is fast, precision is high, favorable reproducibility, strong antijamming capability, it can be achieved that in real time monitoring.
Description
Technical field
The present invention relates to nanoanalysis detection fields, and in particular to one kind is with bovine serum albumin-copper nano-cluster (BSA-
CuNCs it) is used as catalyst, is catalyzed luminol chemiluminescence, quantitatively detects the analysis method of trypsase.
Background technology
Trypsase is one of most important digestive ferment in breaks down proteins, it can be selectively in aminosal by relying
The peptide chain that propylhomoserin or arginic carboxyl are constituted.Trypsase plays key player in controlling exocrine pancreatic function,
Secretion, activation, inhibition and the uneven of cycle of trypsinogen can lead to acute or chronic pancreatic disease.Enzyme linked immunological
Absorption method be trypsase detection a kind of common method, but this method it is costly, time-consuming and usually require that sample size compared with
It is more, and sample preanalysis purification process can lead to the loss of sample, to which the accuracy and reproducibility of testing result can be influenced.
In past many years, the method for the chemiluminescence detection based on bioanalysis is widely used in detecting proteolytic enzyme, egg
White matter, nucleic acid, small molecule etc., while this method also provides a kind of method effectively quickly detecting enzyme.Therefore, exploitation is a kind of
Preparation is easy, can monitor in real time and low-cost trypsase detection method is very necessary and significant.
Chemiluminescence refers to the luminescence phenomenon caused by chemically reacting, and chemiluminescence analysis is to utilize chemiluminescence reaction,
A kind of analysis method that the optical signal sent out when returning ground state by excitation state transition to chemiluminescent substance measures.Luminol
(luminol, i.e. 5- amino -2,3- dihydro-Isosorbide-5-Nitrae-di (hetero) quinoline diketone) is most common chemiluminescent agent.
In recent years, the introducing of emerging nanotechnology and new material significantly improves chemiluminescent detection sensitivity and specificity, and
It is expected to realize Multiple detection.The discovery of metal nano clustered materials causes everybody and widely pays close attention to.Due to quantum confinement effect
It improves so that these extra small metal nanometre clusters have uncommon optics, electrology characteristic, such as Wang G.；Huang T.；
Murray R.W.；Menard L.；J.Am.Chem.Soc.2005,127,812-813；Ramakrishna G.；Varnavski
O.；Kim J.；Lee D.；Goodson T.J.Am.Chem.Soc.2008,130,5032-5033；Zhu M.；Aikens
C.M.；Hollander F.J.；Schatz G.C.；Jin R.J.Am.Chem.Soc.2008,130,5883-5885 etc..As
A kind of new chemical luminescence response unit, noble-metal nanoclusters are remarkably improved the efficiency of luminol chemiluminescence reaction, to opening up
The luminol chemiluminescence reaction system for being unfolded to send out new is of great significance.Have and much joins about nano-particles such as gold, platinum, silver
With luminol chemiluminescence report, such as He Y, He X, Liu X, et al.Dynamically tunable
chemiluminescence of luminol functionalized silver nanoparticles and its
application to protein sensing arrays[J].Anal.Chem.,2014,86:12166-12171 etc., but
The expensive application limited in terms of its biology of noble-metal nanoclusters.
Invention content
The technical problem to be solved in the present invention is to provide one kind using bovine serum albumin-copper nano-cluster as catalyst, catalysis
Luminol chemiluminescence quantitatively detects the analysis method of trypsase, and the detection method is cheap, detection speed is fast, precision
Height, favorable reproducibility, strong antijamming capability are, it can be achieved that monitoring in real time.
Luminol is catalyzed with bovine serum albumin-copper nano-cluster in order to solve the above technical problem, the present invention provides a kind of
The method for detecting trypsase, measuring principle are as follows：Bovine serum albumin-copper nano-cluster can significantly improve Rumi as catalyst
The chemiluminescence of promise-hydrogen peroxide system, trypsase can decompose the bovine serum protein template on copper nano-cluster surface, lead to copper
The change of nano-cluster surface state causes to assemble the reduction so as to cause catalytic activity, and experiment finds the addition of trypsase to Rumi
Promise-hydrogen peroxide-copper nano-cluster chemical luminous system plays apparent inhibiting effect, and the reduction of the chemiluminescence intensity of system
It is linear with the logarithm of trypsinase concentration, the detection of the quantitative analysis to trypsase has been reached based on this.
The detection method includes following step：
S1：Take trypsin solution to be measured, bovine serum albumin-copper nanocluster catalyst, Constant temperature hatch after mixing；
S2：Luminol-hydrogen peroxide detection liquid is added into mixed liquor and carries out chemiluminescence reaction, according to its chemiluminescence
Intensity and the standard curve of foundation, are calculated the concentration of trypsase to be measured.
In the present invention, the preparation method of bovine serum albumin-copper nanocluster catalyst is：By bovine serum albumin(BSA), five water sulphur
Sour copper solution mixing, stirs 5~10min under conditions of 30~40 DEG C, then adjusts pH to alkalinity with sodium hydroxide solution, then
8~16h is stirred under the conditions of 50~60 DEG C, obtains bovine serum albumin-copper nanocluster catalyst.
Further, in step sl, the molar ratio of cupric sulfate pentahydrate and bovine serum albumin(BSA) is (0.5~1):1.
Further, in step sl, pH to 10~12 is adjusted with sodium hydroxide solution.
Further, the bovine serum albumin obtained-copper nanocluster catalyst need to be placed in ultra-pure water and dialyse, not anti-to remove
The impurity answered, it is spare at 2~6 DEG C after dialysis.
Further, a water, dialysis time 40-60h were changed every four hours when dialysis.
In the present invention, the method for building up of standard curve includes：
N group equivalent, various concentration the trypsin solution newly configured is taken, appropriate bovine serum albumin-copper is separately added into
Nanocluster catalyst is separately added into luminol-hydrogen peroxide detection liquid and carries out chemiluminescence reaction after mixing, Constant temperature hatch,
Detect its chemiluminescence intensity；And
According to the decreasing value of the concentration of each group trypsase and corresponding luminous intensity, draw the decreasing value of luminous intensity~
Relation curve between the logarithm of trypsinase concentration, as standard curve.
When establishing standard curve, n values should be the positive integer more than 5, such as n can use 8,10,12.Trypsin solution and
The amount of bovine serum albumin-copper nanocluster catalyst according to circumstances determines, if the amount of trypsin solution is 500 μ L, cow's serum egg
The amount of in vain-copper nanocluster catalyst is 50 μ L.
Further, the concentration range of the trypsin solution of the new configuration is 0.2 μ g/mL~0.6mg/mL.
Further, when detecting sample to be tested and drawing standard curve, the temperature of mixed solution Constant temperature hatch is 30-45
DEG C, brooding time is 0.5h~2h；Luminol-hydrogen peroxide detection architecture is the aqueous solution of luminol and hydrogen peroxide, wherein
A concentration of the 1.0 × 10 of luminol-6~5 × 10-5The pH of M, a concentration of 0.02~0.2M of hydrogen peroxide, system are 9.0.
Detection method according to the present invention can prepare the kit for quantitatively detecting trypsase, to simplify detection
Operation.Further, detection method according to the present invention can also prepare biochip and biosensor, to realize oneself detected
Dynamicization and in real time monitoring, such as biosensor for detecting trypsin amount in blood of human body or urine.
The beneficial effects of the present invention are：
1, detection method of the invention, uses bovine serum albumin-copper nano-cluster for catalyst, catalysis luminol chemistry hair
Light, to improve luminous efficiency.Compared with traditional method using noble-metal nanoclusters as catalyst, the catalyst in the present invention
Bovine serum albumin-copper nano-cluster preparation method is simple, raw material is cheap and easy to get, and catalytic activity is high, and stability is good, reduces detection
Cost.
2, detection method of the invention, anti-impurity interference performance is strong, for the glucose of 10 μ g/L, glucose oxidase,
The Na of lysozyme, urea and 5.0 × 10-4mol/L+、K+、Ca2+、Mg2+Equal interfering substances all have good anti-interference effect
Fruit, to be effectively guaranteed the accuracy of detection.
3, detection method of the invention can be far below the urine of patient for the detection limit of trypsase down to 0.12 μ g/mL
Liquid and the trypsin amount in blood, suitable for detecting the trypsin amount patient urine and blood.
4, detection method of the invention, detection process is pollution-free, speed is fast, easy to operate, favorable reproducibility is, it can be achieved that in real time
Monitoring.
Description of the drawings
Fig. 1, which is luminol-mixed solution of hydrogen peroxide, item existing for (b)/nothing (a) bovine serum albumin-copper nano-cluster
The kinetic curve of luminous intensity under part；
Fig. 2 is that the trypsase of various concentration makees the inhibition of luminol-hydrogen peroxide-copper nano-cluster chemical luminous system
With figure；
Fig. 3 is the canonical plotting for the trypsase that embodiment 1 is established；
Fig. 4 is the result figure that the detection method of the present invention tests the selectivity of trypsase.
Wherein：In figure 3, C is trypsinase concentration, I0It is that pancreas egg is added not add the luminous intensity values of trypsase, I
Luminous intensity values after white enzyme；
In Fig. 4, I0For the chemiluminescence intensity of luminol-hydrogen peroxide-copper nano-cluster system, I is luminol-peroxide
Change hydrogen-chemiluminescence intensity of copper nano-cluster in the presence of various chaff interferents.
Specific implementation mode
The invention will be further described in the following with reference to the drawings and specific embodiments, so that those skilled in the art can be with
It more fully understands the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
In following embodiment, sodium hydroxide, cupric sulfate pentahydrate, luminol, hydrogen peroxide are that analysis is pure, are purchased from traditional Chinese medicines
Chemical reagent Co., Ltd of group；Bovine serum albumin, trypsase are purchased from Sigma-Aldrich companies.
Embodiment 1
A kind of using bovine serum albumin-copper nano-cluster is catalyst luminol chemiluminescence for detecting trypsase
Method includes the following steps：
(1) by bovine serum albumin(BSA), cupric sulfate pentahydrate solution according to molar ratio 1：2 mixing, by mixed liquor in 30 DEG C of conditions
Lower stirring 5min, it is 10 that stirring end is adjusted to pH with sodium hydroxide solution, stirs 8h under the conditions of 50 DEG C, obtains cow's serum egg
In vain-copper nanocluster material；
(2) bovine serum albumin obtained-copper nanocluster material in step (1) is mixed into ultra-pure water the 40h that dialyses, Mei Gesi
Hour changes a water to remove unreacted various impurity, and after dialysis, bovine serum albumin-copper nanocluster catalyst is placed in
It is preserved at 2-6 DEG C for use；
(3) 10 3mL sample cells are taken, the 500 μ L of trypsin solution of fresh configuration, ultimate density difference are separately added into
For 0.4,2.0,10,20,30,46,60,160,400,520,1200 μ g/mL, it is separately added into the ox prepared in 50 μ L steps (2)
Haemocyanin-copper nanocluster catalyst is uniformly mixed, the solution Constant temperature hatch that will be mixed, and incubation temperature is 30 DEG C, when hatching
Between be 0.5h；
(4) luminol is added in 10 sample cells into step (3), and (its ultimate density is 1.10 × 10-4) and peroxide M
The mixed liquor 450uL for changing hydrogen (its ultimate density is 0.45M), is placed in chemiluminescence light path by cuvette rapidly, rapid detection hair
Chemiluminescence intensity of the photo etching at 425nm.
Fig. 1 shows luminol-hydrogen peroxide solution under the conditions of with/without existing for bovine serum albumin-copper nano-cluster
The kinetic curve of luminous intensity.It can be seen from the figure that hydrogen peroxide oxidation luminol produces very weak chemiluminescence
(a), with the addition of bovine serum albumin-copper nano-cluster, (b) is remarkably reinforced in the luminous intensity of mixed liquor, and two luminescence systems
Maximum emission wavelength is consistent, and is 425nm, this explanation is under the catalytic action of bovine serum albumin-copper nano-cluster, Rumi
The chemical luminophor of promise-mixed solution of hydrogen peroxide is still the oxidation product of luminol, i.e. the 3- amino neighbours benzene two of excitation state
Formate anion (3-APA*), bovine serum albumin-copper nano-cluster only play catalytic action, do not generate new illuminator.
Fig. 2 shows various concentration trypsase is added in luminol-hydrogen peroxide-copper nano-cluster chemical luminous system
Afterwards, the luminous intensity variations figure of system.It can be seen from the figure that trypsase is to luminol-hydrogen peroxide-copper nano-cluster chemistry
Luminescence system plays apparent inhibiting effect.
Using logC (C is trypsinase concentration) for abscissa, with I0-I(I0Not add the luminous intensity values of trypsase, I
For the luminous intensity values after trypsase are added) it is ordinate, according to logC and I0The correspondence of-I draws standard curve, knot
Fruit is as shown in Figure 3.As can be seen that in 0.2 μ g/mL~0.6mg/mL concentration ranges, the variation of luminous intensity and trypsase
The logarithm of concentration is linear, and the quantitative detecting analysis to carrying out trypsase is realized based on this.
Embodiment 2
A kind of using bovine serum albumin-copper nano-cluster is catalyst luminol chemiluminescence for detecting trypsase
Method includes the following steps：
(1) by bovine serum albumin(BSA), cupric sulfate pentahydrate solution according to molar ratio 3：4 mixing, by mixed liquor in 35 DEG C of conditions
Lower stirring 7.5min, stirring end adjust pH to 11 with sodium hydroxide solution, stir 12h under the conditions of 55 DEG C, obtain cow's serum
Albumen-copper nanocluster material；
(2) bovine serum albumin obtained-copper nanocluster material in step (1) is mixed into ultra-pure water the 50h that dialyses, Mei Gesi
Hour changes a water to remove unreacted various impurity, and after dialysis, bovine serum albumin-copper nanocluster catalyst is placed in
It is preserved at 2-6 DEG C for use；
(3) 10 3mL sample cells are taken, the 500 μ L of trypsin solution of fresh configuration, ultimate density difference are separately added into
For 0.4,2.0,10,20,30,46,60,160,400,520,1200 μ g/mL, then it is separately added into system in 50 μ L steps (2) of equivalent
Standby bovine serum albumin-copper nanocluster catalyst is uniformly mixed, the solution Constant temperature hatch that will be mixed, incubation temperature 37.5
DEG C, brooding time 1.25h；
(4) luminol is added in 10 sample cells into step (3), and (its ultimate density is 1.10 × 10-4) and peroxide M
The mixed liquor 450uL for changing hydrogen (its ultimate density is 0.45M), is placed in chemiluminescence light path by cuvette rapidly, rapid detection hair
Chemiluminescence intensity of the photo etching at 425nm.
Embodiment 3
A kind of using bovine serum albumin-copper nano-cluster is catalyst luminol chemiluminescence for detecting trypsase
Method includes the following steps：
(1) by bovine serum albumin(BSA), cupric sulfate pentahydrate solution according to the ratio between certain amount of substance mix, by mixed liquor in
10min is stirred under the conditions of 40 DEG C, stirring end adjusts pH to 12 with sodium hydroxide solution, 16h is stirred under the conditions of 50-60 DEG C,
Obtain bovine serum albumin-copper nanocluster material；
(2) bovine serum albumin obtained-copper nanocluster material in step (1) is mixed into ultra-pure water the 60h that dialyses, Mei Gesi
Hour changes a water to remove unreacted various impurity, and after dialysis, bovine serum albumin-copper nanocluster catalyst is placed in
It is preserved at 2-6 DEG C for use；
(3) 10 3mL sample cells are taken, the 500 μ L of trypsin solution of fresh configuration, ultimate density difference are separately added into
For 0.4,2.0,10,20,30,46,60,160,400,520,1200 μ g/mL, it is separately added into 50 μ L steps (2) of equivalent and prepares
Bovine serum albumin-copper nanocluster catalyst, be uniformly mixed, the solution Constant temperature hatch that will be mixed, incubation temperature be 45 DEG C, incubate
The change time is 2h,
(4) luminol is added in 10 sample cells into step (3), and (its ultimate density is 1.10 × 10-4) and peroxide M
The mixed liquor 450uL for changing hydrogen (its ultimate density is 0.45M), is placed in chemiluminescence light path by cuvette rapidly, rapid detection hair
Chemiluminescence intensity of the photo etching at 425nm.
Embodiment 4
It is specific as follows to the selectivity experiment of trypsase：
Take respectively the trypsase glucose of 10 μ g/L of fresh configuration, glucose oxidase, lysozyme, urea and
5.0×10-4The Na of mol/L+、K+、Ca2+、Mg2+Solution is separately added into appropriate bovine serum albumin-copper nanocluster catalyst, through mixed
It closes, after Constant temperature hatch, is separately added into luminol-hydrogen peroxide detection liquid and carries out chemiluminescence reaction, it is strong to detect its chemiluminescence
Degree, acquired results are as shown in Figure 4.
As can be drawn from Figure 4, the glucose of 10 μ g/L, glucose oxidase, lysozyme, urea and 5.0 × 10- 4The Na of mol/L+、K+、Ca2+、Mg2+Do not have a great impact to the chemiluminescence intensity of system, and the pancreas egg of 10 μ g/L is added
White enzyme can be such that the chemiluminescence intensity of system is substantially reduced, and illustrate that detection architecture has good selectivity trypsase, should
Detection architecture is for when detecting trypsase, having very strong anti-interference ability, to ensure that the accuracy of detection.
Embodiment described above is only to absolutely prove preferred embodiment that is of the invention and being lifted, protection model of the invention
It encloses without being limited thereto.Those skilled in the art on the basis of the present invention made by equivalent substitute or transformation, in the present invention
Protection domain within.Protection scope of the present invention is subject to claims.
Claims (10)
1. the method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence.
2. detecting trypsase as described in claim 1 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that including：
S1：Take trypsin solution to be measured, bovine serum albumin-copper nanocluster catalyst, Constant temperature hatch after mixing；
S2：Luminol-hydrogen peroxide detection liquid is added into mixed liquor and carries out chemiluminescence reaction, according to its chemiluminescence intensity
And the standard curve established, the concentration of trypsase to be measured is calculated.
3. detecting trypsase as described in claim 1 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that in step sl, the preparation method of bovine serum albumin-copper nanocluster catalyst is：By bovine serum albumin
In vain, cupric sulfate pentahydrate solution mixes, and 5~10min is stirred under conditions of 30~40 DEG C, then adjusts pH with sodium hydroxide solution
To alkalinity, 8~16h is stirred under the conditions of 50~60 DEG C, obtains bovine serum albumin-copper nanocluster catalyst.
4. detecting trypsase as claimed in claim 3 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that in step sl, the molar ratio of cupric sulfate pentahydrate and bovine serum albumin(BSA) is (0.5~1):1.
5. detecting trypsase as claimed in claim 3 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that in step sl, pH to 10~12 is adjusted with sodium hydroxide solution.
6. detecting trypsase as claimed in claim 3 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that obtained bovine serum albumin-copper nanocluster catalyst need to be placed in ultra-pure water and dialyse, not anti-to remove
The impurity answered, it is spare at 2~6 DEG C after dialysis.
7. detecting trypsase as described in claim 1 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that the method for building up of standard curve includes：
N group equivalent, various concentration the trypsin solution newly configured is taken, appropriate bovine serum albumin-copper nanometer is separately added into
Cluster catalyst is separately added into luminol-hydrogen peroxide detection liquid and carries out chemiluminescence reaction, detection after mixing, Constant temperature hatch
Its chemiluminescence intensity；And
According to the decreasing value of the concentration of each group trypsase and corresponding luminous intensity, decreasing value~pancreas egg of luminous intensity is drawn
Relation curve between the logarithm of white enzyme concentration, as standard curve.
8. detecting trypsase as claimed in claim 7 with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that the concentration range of the trypsin solution of the new configuration is 0.2 μ g/mL~0.6mg/mL.
9. as claimed in claim 1 or 7 detect trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
Method, which is characterized in that the temperature of Constant temperature hatch be 30-45 DEG C, the time be 0.5h~2h.
10. as claimed in claim 1 or 7 detect tryptose with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence
The method of enzyme, which is characterized in that luminol-hydrogen peroxide detection liquid is the aqueous solution of luminol and hydrogen peroxide, pH 9.0,
Wherein a concentration of the 1.0 × 10 of luminol-6~5 × 10-5M, a concentration of 0.02~0.2M of hydrogen peroxide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810331532.1A CN108535240A (en) | 2018-04-13 | 2018-04-13 | The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810331532.1A CN108535240A (en) | 2018-04-13 | 2018-04-13 | The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108535240A true CN108535240A (en) | 2018-09-14 |
Family
ID=63480430
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810331532.1A Pending CN108535240A (en) | 2018-04-13 | 2018-04-13 | The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108535240A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111579548A (en) * | 2020-05-20 | 2020-08-25 | 重庆师范大学 | Luminol-gallium nano assembly and preparation method and application thereof |
| CN113433100A (en) * | 2021-05-25 | 2021-09-24 | 上海市公共卫生临床中心 | Plasma tryptophan and albumin joint detection method based on photochemical reaction of DNA synthesized silver nanoclusters and tryptophan |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1553962A (en) * | 2001-08-10 | 2004-12-08 | 阿赫姆生物系统公司 | System for detecting protease |
| CN102654501A (en) * | 2012-05-21 | 2012-09-05 | 江苏省原子医学研究所 | Method for detecting optically-excited chemiluminiscence of pepsinogen II and reagent kit |
| CN104655615A (en) * | 2014-07-08 | 2015-05-27 | 青岛科技大学 | Biological sensor based on CuS nano-particle cation exchange enhanced chemiluminescence |
-
2018
- 2018-04-13 CN CN201810331532.1A patent/CN108535240A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1553962A (en) * | 2001-08-10 | 2004-12-08 | 阿赫姆生物系统公司 | System for detecting protease |
| CN102654501A (en) * | 2012-05-21 | 2012-09-05 | 江苏省原子医学研究所 | Method for detecting optically-excited chemiluminiscence of pepsinogen II and reagent kit |
| CN104655615A (en) * | 2014-07-08 | 2015-05-27 | 青岛科技大学 | Biological sensor based on CuS nano-particle cation exchange enhanced chemiluminescence |
Non-Patent Citations (3)
| Title |
|---|
| SHUANGJIAO XU 等: "Luminol chemiluminescence enhanced by copper nanoclusters and its analytical application", 《RSC ADVANCES》 * |
| XIAOYING YOU 等: "Gold nanoclusters-based chemiluminescence resonance energy transfer method for sensitive and label-free detection of trypsin", 《TALANTA》 * |
| 吴秀明 等: "铜纳米簇催化鲁米诺化学发光体系的研究及应用", 《化学研究与应用》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111579548A (en) * | 2020-05-20 | 2020-08-25 | 重庆师范大学 | Luminol-gallium nano assembly and preparation method and application thereof |
| CN113433100A (en) * | 2021-05-25 | 2021-09-24 | 上海市公共卫生临床中心 | Plasma tryptophan and albumin joint detection method based on photochemical reaction of DNA synthesized silver nanoclusters and tryptophan |
| CN113433100B (en) * | 2021-05-25 | 2022-09-27 | 上海市公共卫生临床中心 | Plasma tryptophan and albumin joint detection method based on photochemical reaction of DNA synthesized silver nanoclusters and tryptophan |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102998413B (en) | The purposes of gold-core/platinum-shell nano bar analogue enzyme solution and be detected the method for hydrogen oxide, glucose and cholesterol | |
| Jiang et al. | Electrochemiluminescence of luminol enhanced by the synergetic catalysis of hemin and silver nanoparticles for sensitive protein detection | |
| Lin et al. | Luminol chemiluminescence in unbuffered solutions with a cobalt (II)− ethanolamine complex immobilized on resin as catalyst and its application to analysis | |
| CN102507543B (en) | Method for enhancing luminol chemoluminescence using copper oxide nanoparticles | |
| CN110186910B (en) | Double-inhibition electrochemiluminescence sensor and method for measuring glyphosate | |
| Xie et al. | Co 3 O 4 nanoparticles-enhanced luminol chemiluminescence and its application in H 2 O 2 and glucose detection | |
| CN109092364A (en) | A kind of copper metal organic backbone analogue enztme material and its preparation and application | |
| CN102967595B (en) | Chemical luminous sensor and method for detecting glucose by employing chemical luminous sensor | |
| CN104459157B (en) | A kind of preparation method of the immunosensor based on ferrocene-cuprous oxide cubic nanometer Framework Mark and application | |
| CN108801998B (en) | A method of the ratio fluorescent probe in detecting choline based on copper nano-cluster compound | |
| CN105891189B (en) | A kind of copper ion detection kit and its application | |
| CN110455786B (en) | Based on CeO2@SnS2Preparation method of luminol-promoted electrochemiluminescence sensor | |
| CN106940315A (en) | On Detection of Organophosphorus Pesticide and kit | |
| CN109342529B (en) | Non-enzymatic glucose sensor and preparation method thereof | |
| CN108535240A (en) | The method for detecting trypsase with bovine serum albumin-copper nano-cluster catalysis luminol chemiluminescence | |
| Shao et al. | Recent advances in enzyme-enhanced immunosensors | |
| Zhang et al. | Silicon quantum dot involved luminol chemiluminescence and its sensitive detection of dopamine | |
| CN103512855B (en) | The detection method of reduced glutathione | |
| CN102798627B (en) | Method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as catalyst | |
| CN106546585B (en) | The detection method and detection kit of mercury ion, total mercury and organic mercury | |
| Li et al. | A fluorescent sensor based on ovalbumin-modified Au nanoclusters for sensitive detection of ascorbic acid | |
| Zou et al. | Stimuli-responsive colorimetric sensor based on bifunctional pyrophosphate-triggered controlled release and enhancing activity of CoOOH nanozyme | |
| CN106770215B (en) | A kind of preparation method of iron cobalt magnetic Nano sensor of multifunction and products thereof and application | |
| CN105021580A (en) | 17beta-estradiol detection method | |
| CN108896750A (en) | A kind of preparation method and purposes of BSA-Au/Ag NCs/OPD/HRP proportional-type fluorescent optical sensor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180914 |
|
| RJ01 | Rejection of invention patent application after publication |