CN105277537B - A method of detection enzymatic activity and chemiluminescence reaction substrate performance - Google Patents
A method of detection enzymatic activity and chemiluminescence reaction substrate performance Download PDFInfo
- Publication number
- CN105277537B CN105277537B CN201510657942.1A CN201510657942A CN105277537B CN 105277537 B CN105277537 B CN 105277537B CN 201510657942 A CN201510657942 A CN 201510657942A CN 105277537 B CN105277537 B CN 105277537B
- Authority
- CN
- China
- Prior art keywords
- substrate
- chemiluminescence
- reaction substrate
- chemiluminescence reaction
- enzyme solutions
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses it is a kind of detection enzymatic activity and chemiluminescence reaction substrate performance method, including glow cup, chemiluminescence reaction substrate A, chemical reaction luminous substrate B, with the chemiluminescence reaction substrate A, chemically react luminous substrate B match the enzyme solutions of detection, lighting apparatus；The chemiluminescence reaction substrate A, the foundation for chemically reacting luminous substrate B and the enzyme solutions luminescence system detect the performance of the enzyme solutions activity or the chemiluminescence reaction substrate according to the luminescence system of the foundation.Method of the invention can fast and effeciently a variety of enzymes that can cause chemiluminescence phenomenon of Accurate Determining activity and concentration, and it can be particularly suitable for two kinds of chemiluminescent substrates according to the performance of known concentration enzyme solutions rapidly and accurately evaluating chemical luminous substrate and be compared.
Description
Technical field
The present invention relates to bioanalytical chemistry technical fields, and in particular to a kind of detection enzymatic activity and chemiluminescence reaction bottom
The method of physical performance.
Background technique
Chemiluminescence is that substance is carrying out a kind of light radiation phenomenon adjoint in chemical reaction process, and luminescence mechanism is anti-
Answer the Cucumber in system to absorb the energy of reaction release and by ground state transition to excitation state, when returning to ground state from excitation state
Energy is released in the form of light radiation, generates luminescence phenomenon.Chemiluminometry is then according to a certain moment chemistry
Luminous intensity or chemiluminescence total amount determine a kind of micro and trace analysis method of respective components content in reaction, have Gao Ling
The features such as sensitivity, the range of linearity are wide, equipment is simple, easy to operate, easy to automate and analysis is quick.In recent years, with stream
The combination of the methods of dynamic injection, electrochemistry, high performance liquid chromatography and Capillary Electrophoresis, so that chemiluminometry is answered extensively
For fields such as Pharmaceutical Analysis, clinical analysis, environmental analysis and material analysis.Chemiluminescent substrate research and development, selection, stability are surveyed
In the work such as examination, effective performance evaluation means have been required.General enterprises are all using ELISA or self-control kit pilot scale
Mode is evaluated.The problems such as it is too many all to there is influence factor in both methods, time-consuming and laborious, at high cost.
How quickly, quasi- the concentration of enzyme and active (or enzyme activity) are the projects paid high attention in field of biology,
The concentration and activity for really measuring enzyme are to realize the basis of proper use of enzyme and enzyme preparation.The survey that major part producer uses at present
The method of determining is electrophoresis, immunization, the methods of bioanalysis.The generally existing testing cost of these methods is high, and the period is long, not accurate enough
The disadvantages of.
Summary of the invention
Based on this, it is necessary to provide one kind quickly, simplicity, directly detection enzymatic activity and chemiluminescence reaction substrate performance
Method is at least one of to solve the above technical problems.
A method of detection enzymatic activity and chemiluminescence reaction substrate performance, comprising: glow cup, chemiluminescence reaction bottom
Object A, the enzyme for chemically reacting luminous substrate B, matching detection with the chemiluminescence reaction substrate A, chemical reaction luminous substrate B
Solution, lighting apparatus and the chemiluminescence reaction substrate A, chemical reaction luminous substrate B and the enzyme solutions luminescence system
It establishes, the enzyme solutions activity or enzyme concentration or the chemiluminescence reaction bottom is detected according to the luminescence system of the foundation
The performance of object；
The foundation of the luminescence system the following steps are included:
1) the chemiluminescence reaction substrate A and chemical reaction luminous substrate B are modulated be uniformly mixed by a certain percentage, and
It is placed in the glow cup, it is spare；
2) enzyme solutions are diluted to a series of different concentration；
3) by the enzyme solutions diluted described in step 2 as in the glow cup with the chemiluminescence reaction substrate A and
The mixed liquor of chemical reaction luminous substrate B is sufficiently mixed, and is placed into the measurement for carrying out light signal strength in the lighting apparatus, is obtained
Optical signals intensity；
4) light signal strength according to when a series of various concentrations that step 3 obtains establish the enzyme solutions concentration or
Linear relationship between person and chemiluminescence signal intensity.
Further, detection enzyme solutions activity the following steps are included:
1) the chemiluminescence reaction substrate A and chemical reaction luminous substrate B are modulated be uniformly mixed by a certain percentage, and
It is placed in the glow cup, it is spare；
2) enzyme solutions are diluted to certain concentration；
3) by the enzyme solutions diluted described in step 2 as in the glow cup with the chemiluminescence reaction substrate A and
The mixed liquor of chemical reaction luminous substrate B is sufficiently mixed, and is placed into the measurement for carrying out light signal strength in the lighting apparatus, is obtained
Optical signals intensity X；
4) the light signal strength X that step 3 obtains is compared with the luminescence system of the foundation, calculates the enzyme solutions
Activity or concentration or the chemiluminescence reaction substrate performance.
Further, the chemiluminescence reaction substrate A and chemical reaction luminous substrate B can be for can be molten with the enzyme
Liquid generates chemiluminescent any chemiluminescence reaction substrate, including and is not limited to luminol-hydrogen peroxide reaction substrate, phosphoric acid
Or reaction substrate, the chemical reaction substrate containing ruthenium of phosphoric acid ester.
Further, the enzyme solutions be can with the chemiluminescence reaction substrate A and chemically react luminous substrate B into
Row chemiluminescence reaction and the enzyme for generating optical signal, including and it is not limited to catalase, horseradish peroxidase or alkaline phosphorus
Sour enzyme.
Further, the chemiluminescence reaction substrate A can be luminol luminescent solution A liquid, and the chemical reaction shines
Substrate B can be luminol luminescent solution B liquid.
Further, the luminol luminescent solution A liquid includes luminol, Tris- buffer, the luminol luminescent solution B
Liquid includes peroxide ingredient.
Further, the luminol luminescent solution A liquid and the luminol luminescent solution B liquid are modulated mixed according to the ratio of 1:1
It closes.
Further, the material of the glow cup is the plastics to visible light without absorption.
Further, the inner cavity of the glow cup is cuboid, and the uniform wall thickness of the glow cup.
Further, the lighting apparatus is the equipment that can measure light signal strength, including and be not limited to camera,
Photocell, photomultiplier tube, chemical luminescent detecting equipment.
Further, the lighting apparatus is CR-0302 type optical signal reading apparatus.
Beneficial effects of the present invention:
(1) method provided by the invention that enzymatic activity and chemiluminescence reaction substrate performance are detected based on chemoluminescence method
Can fast and effeciently a variety of enzymes that can cause chemiluminescence phenomenon of Accurate Determining activity and concentration.This method detects only every time
2-3 minutes time is needed, it is at low cost.It is very suitable to enzyme preparation manufacturing enterprise or is carried out using performance of the enterprise to enzyme preparation quick
Evaluation.
(2) method provided by the invention that enzymatic activity and chemiluminescence reaction substrate performance are detected based on chemoluminescence method
As long as the performance of the enzyme solutions rapidly and accurately evaluating chemical luminous substrate using known concentration.It is particularly suitable for two
Kind chemiluminescent substrate is compared.
Detailed description of the invention
Luminous photo legend when Fig. 1 is different horseradish peroxidase concentration.
Linear relationship curve of the Fig. 2 between chemiluminescence signal intensity and the concentration of enzyme.
Specific embodiment
A kind of method of detection enzymatic activity and chemiluminescence reaction substrate performance provided by the invention, including glow cup, change
Learn luminescence-producing reaction substrate A, chemical reaction luminous substrate B, with the chemiluminescence reaction substrate A, chemically react luminous substrate B phase
Cooperate enzyme solutions, lighting apparatus and the chemiluminescence reaction substrate A, chemical reaction luminous substrate B and the enzyme of detection molten
The foundation of liquid luminescence system detects the enzyme solutions activity or the chemiluminescence reaction according to the luminescence system of the foundation
The performance of substrate；
The foundation of the luminescence system the following steps are included:
1) the chemiluminescence reaction substrate A and chemical reaction luminous substrate B are modulated be uniformly mixed by a certain percentage, and
It is placed in the glow cup, it is spare；
2) enzyme solutions are diluted to a series of different concentration；In detail, using the enzyme solutions of known concentration as ginseng
According to enzyme solutions, enzyme solutions to be measured are measured and are evaluated.Enzyme solutions to be measured are made into a series of enzyme solutions of various concentrations, it is dense
Degree range is 1pg/ml-1ug/ml, preferably 1ng/ml-100ng/ml, the chemiluminescence light that the enzyme solutions of various concentration generate
Signal strength is different, and in a linear relationship with the concentration of enzyme solutions.
3) by the enzyme solutions diluted described in step 2 be placed in the glow cup with the chemiluminescence reaction substrate A and
The mixed liquor of chemical reaction luminous substrate B is sufficiently mixed, and is placed into the measurement for carrying out light signal strength in the lighting apparatus, is obtained
Optical signals intensity；
4) light signal strength according to when a series of various concentrations that step 3 obtains establish the enzyme solutions concentration or
Linear relationship between person and chemiluminescence signal intensity.
Further, detection enzyme solutions activity the following steps are included:
1) the chemiluminescence reaction substrate A and chemical reaction luminous substrate B are modulated be uniformly mixed by a certain percentage, and
It is placed in the glow cup, it is spare；
2) enzyme solutions are diluted to certain concentration；
3) by the enzyme solutions diluted described in step 2 as in the glow cup with the chemiluminescence reaction substrate A and
The mixed liquor of chemical reaction luminous substrate B is sufficiently mixed, and is placed into the measurement for carrying out light signal strength in the lighting apparatus, is obtained
Optical signals intensity X；
4) the light signal strength X that step 3 obtains is compared with the luminescence system of the foundation, calculates the enzyme solutions
Activity or concentration or the chemiluminescence reaction substrate performance.
Preferably, the chemiluminescence reaction substrate A and chemical reaction luminous substrate B can be for can be with the enzyme solutions
Generate chemiluminescent any chemiluminescence reaction substrate, including and be not limited to luminol-hydrogen peroxide reaction substrate, phosphoric acid or
The reaction substrate of phosphoric acid ester, the chemical reaction substrate containing ruthenium.
Further, the enzyme solutions be can with the chemiluminescence reaction substrate A and chemically react luminous substrate B into
Row chemiluminescence reaction and the enzyme for generating optical signal, including and it is not limited to catalase, horseradish peroxidase or alkaline phosphorus
Sour enzyme.
Further, the chemiluminescence reaction substrate A can be luminol luminescent solution A liquid, and the chemical reaction shines
Substrate B can be luminol luminescent solution B liquid.The chemiluminescence reaction substrate A that the present invention uses be also possible to different luminol or
It derives class object.The luminol luminescent solution A liquid includes luminol and Tris- buffer, and the luminol luminescent solution B liquid includes
Peroxide ingredient.Luminol luminescent solution A liquid and the luminol luminescent solution B liquid are modulated according to the ratio of 1:1 when allotment mixing
Mixing.
Preferably, the glow cup is used to visible light substantially without the plastic production of absorption.The inner cavity of glow cup is rectangular
The bodily form, and the uniform wall thickness of the glow cup.The inner cavity capacity of glow cup is 300ul or so.Entire inner cavity is in cuboid, is made
The thickness of each position solution is without significant difference.Had using the glow cup and is shone uniformly, it is easy to operate, measure the advantages that accurate.Hair
The inner cavity specific size of light cup is long 20mm wide 5mm thickness 3mm.Of course, the interior chamber size of glow cup can according to need progress
Adjustment, it is preferred that in homogeneous thickness to reduce evaluated error.
Preferably, the lighting apparatus is the equipment that can measure light signal strength, including and is not limited to camera, light
Battery, photomultiplier tube, chemical luminescent detecting equipment.Of course, such as high sense of other equipment for being able to detect light signal strength
Degree camera system can be applied.The lighting apparatus used in an embodiment of the present invention is CR-0302 type optical signal reading apparatus.
Present invention could apply to detect enzyme solutions activity,
Specifically, for 100, the activity of enzymatic reagent to be measured is measured and be evaluated referring to the activity of enzymatic reagent.It will ginseng
The solution of same concentrations, concentration range 1pg/ml-1ug/ml, preferably 1ng/ml- are made into according to enzymatic reagent and enzymatic reagent to be measured
100ng/ml, different enzymatic reagents have most suitable concentration, and preferable concentration is that the chemiluminescence optical signal generated is about optical signal
The half or so of detection device upper limit of detection.
A certain amount of chemiluminescent substrate is prepared in glow cup in advance according to its requirement.Then thereto
A certain amount of enzyme solutions are added.Chemiluminescence intensity is measured with lighting apparatus after being sufficiently mixed.Record chemiluminescence signal intensity.
Enzymatic activity is calculated according to the following formula:
Enzymatic activity=100 × enzyme to be measured signal strength/reference enzyme signal strength
Present invention can also apply to detect enzyme solutions concentration.
Specifically, when being measured to the effective concentration of enzyme, a series of differences first are prepared using the enzyme solutions of known concentration
The enzyme solutions of concentration.The concentration of enzyme solutions be formulated as 0ng/ml, 1ng/ml, 2ng/ml, 3ng/ml, 5ng/ml, 10ng/ml,
18ng/ml,20ng/ml,25ng/ml.Then the chemistry of the enzyme solutions and same dose that measure various concentration in aforementioned manners is sent out
Light reaction substrate A and chemiluminescence reaction substrate B carry out the light signal strength of sending when chemiluminescence reaction, draw chemiluminescence
Relation curve between signal strength and enzyme solutions concentration.As shown in Figure 1, shining when being different horseradish peroxidase concentration
Photo legend.Wherein 1 be the horseradish peroxidase solution that concentration is 0ng/ml luminous photo, 2 be concentration be 1ng/ml's
The luminous photo of horseradish peroxidase solution, 3 be the luminous photo for the horseradish peroxidase solution that concentration is 10ng/ml.
Chemiluminescence signal intensity as shown in Figure 2 and the concentration of enzyme present good linear.Then same to the enzyme solutions of concentration to be measured
Chemiluminescent measurement is carried out, according to the pass between the luminous signal strength chemical luminous signal intensity measured and enzyme solutions concentration
It is that curve compares, extrapolates the effective concentration of enzyme.
The present invention can also evaluate the performance of chemiluminescent substrate.
Specifically, by this method can the performance to chemiluminescent substrate accurately evaluated, and not will receive examination
The interference of the other factors such as agent box performance.Under the action of same amount of enzyme preparation, with shining referring to chemiluminescent substrate
Performance is 100, and the signal that chemiluminescent substrate to be measured is generated can evaluate chemistry to be measured compared with the signal referring to substrate
The performance of luminous substrate.Such as " the SuperSignal ELISA Femto maximum sensitivity bottom produced with Sai Mofei company, the U.S.
Object " is referring to substrate.The light signal strength that the chemiluminescent substrate used in the present invention generates flies Products to match to write from memory
95%.Then the performance scores of the chemiluminescent substrate are 95.
Embodiment one: the detection of enzymatic activity.
Use chemiluminescent substrate: SF-2 type luminol ultra-sensitive chemical luminous substrate,
Use lighting apparatus: CR-0302 type optical signal reading apparatus
Glow cup 1 is taken, 100ul luminol luminescent solution A liquid and 100ul luminol luminescent solution are separately added into glow cup
B liquid.The horseradish peroxidase solution to be measured that 10ul has diluted then is added, enzyme concentration 10ng/ml is carried out immediately after mixing
It takes pictures, sensitive time 30 seconds.Pay attention to avoiding bubble.Obtaining signal value is 1068.Same method measurement is referring to horseradish peroxidase
Enzyme solutions (the silent winged product of U.S.'s match), obtaining the activity value that signal value is 1100. tested horseradish peroxidases is 97.
Embodiment two: the detection of enzyme solutions concentration.
Use chemiluminescent substrate: SF-2 type luminol ultra-sensitive chemical luminous substrate
Use lighting apparatus: CR-0302 type optical signal reading apparatus
100ul luminol luminescent solution A liquid and 100ul luminol luminescent solution B liquid are separately added into glow cup.Then it is added
10ul various concentration horseradish peroxidase solution (each enzyme concentration is 100,80,60,40,20,10,5,2ng/ml), each sample
It takes pictures immediately after mixing, sensitive time 30 seconds.Relation curve is drawn according to each signal value result.
It to enzyme solutions to be measured, is equally operated, after obtaining signal value, the dense of the enzyme solutions is read from working curve
Degree.
Embodiment three: chemiluminescent substrate performance evaluation.
Use enzyme: horseradish peroxidase, concentration 10ng/ml.
Referring to chemiluminescent substrate: SuperSignal ELISAFemto maximum sensitivity substrate,
Test object: SF-2 type luminol ultra-sensitive chemical luminous substrate,
Use lighting apparatus: CR-0302 type optical signal reading apparatus
Glow cup 1 is taken, homemade 100ul luminol luminescent solution A liquid and 100ul luminol are separately added into glow cup
Luminescent solution B liquid.The horseradish peroxidase solution that 10ul has diluted then is added, enzyme concentration 10ng/ml is carried out immediately after mixing
It takes pictures, sensitive time 30 seconds.Pay attention to avoiding bubble.Obtaining signal value is 1072. same using the chemiluminescent substrate progress of reference
The measurement of sample, obtaining signal value is 1128.The performance number for then making chemiluminescent substrate by oneself is 95.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (5)
1. a kind of method of detection enzymatic activity and chemiluminescence reaction substrate performance, which is characterized in that including glow cup, chemistry hair
Light reaction substrate A, chemical reaction luminous substrate B, it is matched with the chemiluminescence reaction substrate A, chemical reaction luminous substrate B
Enzyme solutions, the lighting apparatus of detection；The chemiluminescence reaction substrate A, chemical reaction luminous substrate B and the enzyme solutions shine
The foundation of system；
The detection enzyme solutions activity or the performance of the chemiluminescence reaction substrate the following steps are included:
1) the chemiluminescence reaction substrate A and chemical reaction luminous substrate B are modulated be uniformly mixed by a certain percentage, be placed in
It is spare in the glow cup；
2) enzyme solutions are diluted to certain concentration；
3) the step 2) enzyme solutions diluted are placed in the glow cup and the chemiluminescence reaction substrate A and chemistry
The mixed liquor of reaction luminous substrate B is sufficiently mixed, and is placed into the measurement for carrying out light signal strength in the lighting apparatus, is obtained light
Signal strength X；
4) performance of the active or described chemiluminescence reaction substrate of the enzyme solutions is calculated；
Wherein, the chemiluminescence reaction substrate A and chemical reaction luminous substrate B be can directly with the enzyme solutions generationization
Luminous any chemiluminescence reaction substrate is learned, including the reaction of luminol-hydrogen peroxide reaction substrate, phosphoric acid or phosphoric acid ester
Substrate, the chemical reaction substrate containing ruthenium；The enzyme solutions be can directly with the chemiluminescence reaction substrate A and chemical reaction
Luminous substrate B carries out chemiluminescence reaction and generates the enzyme of optical signal, including catalase, horseradish peroxidase or alkalinity
Phosphatase；
Wherein, the chemiluminescence reaction substrate A is luminol luminescent solution A liquid, and the chemical reaction luminous substrate B is luminol
Luminescent solution B liquid；The luminol luminescent solution A liquid includes luminol and Tris- buffer components, the luminol luminescent solution B liquid
Including peroxide ingredient；The luminol luminescent solution A liquid and the luminol luminescent solution B liquid are modulated mixed according to the ratio of 1:1
It closes；
Wherein, for 100, the activity of enzymatic reagent to be measured is measured and be evaluated, is counted according to the following formula referring to the activity of enzymatic reagent
Calculate the enzymatic activity of enzymatic reagent to be measured:
Enzymatic activity=100 × enzyme to be measured signal strength/reference enzyme signal strength.
2. the method for detection enzymatic activity according to claim 1 and chemiluminescence reaction substrate performance, which is characterized in that institute
The material of glow cup is stated as the plastics to visible light without absorption.
3. the method for detection enzymatic activity according to claim 2 and chemiluminescence reaction substrate performance, which is characterized in that institute
The inner cavity for stating glow cup is cuboid, the uniform wall thickness of the glow cup.
4. the method for detection enzymatic activity according to claim 1 and chemiluminescence reaction substrate performance, which is characterized in that institute
The lighting apparatus stated is the equipment that can measure light signal strength, including camera, photocell, photomultiplier tube, chemiluminescence
Sensing equipment.
5. the method for detection enzymatic activity according to claim 4 and chemiluminescence reaction substrate performance, which is characterized in that institute
The lighting apparatus stated is CR-0302 type optical signal reading apparatus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510657942.1A CN105277537B (en) | 2015-10-12 | 2015-10-12 | A method of detection enzymatic activity and chemiluminescence reaction substrate performance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510657942.1A CN105277537B (en) | 2015-10-12 | 2015-10-12 | A method of detection enzymatic activity and chemiluminescence reaction substrate performance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105277537A CN105277537A (en) | 2016-01-27 |
| CN105277537B true CN105277537B (en) | 2019-04-16 |
Family
ID=55146921
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510657942.1A Active CN105277537B (en) | 2015-10-12 | 2015-10-12 | A method of detection enzymatic activity and chemiluminescence reaction substrate performance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105277537B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114184584B (en) * | 2021-12-06 | 2024-06-04 | 山东航空学院 | Enzyme determination micelle liquid and marine natural product antioxidant activity evaluation method |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1474185A (en) * | 2003-07-30 | 2004-02-11 | 中国药科大学 | Luminol chemiluminescence immunological analysis detecting method for cardiac muscle calcium protein |
| CN1687751A (en) * | 2005-05-25 | 2005-10-26 | 段虎 | High stable enhanced chemiluminescence substrate |
| CN1756848A (en) * | 2003-04-17 | 2006-04-05 | 坎布里克斯生物科学诺丁汉有限公司 | Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity |
| CN1912589A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence measuring method of urea in serum |
| CN101218354A (en) * | 2005-05-13 | 2008-07-09 | 珀金埃尔默生命与分析科学有限公司 | Use of ammonium and phosphate ions for improving the detection of luciferase |
| CN101639446A (en) * | 2009-09-07 | 2010-02-03 | 福建省洪诚生物药业有限公司 | Method for detecting blood lactic acid in vitro by using chemiluminescence method |
| CN101639445A (en) * | 2009-09-07 | 2010-02-03 | 福建省洪诚生物药业有限公司 | Method for detecting blood pyruvate in vitro by using chemiluminescence method |
| CN102809541A (en) * | 2012-05-14 | 2012-12-05 | 浙江省海洋开发研究院 | Determination method of enzyme activity of collagenase |
| CN103063657A (en) * | 2012-12-24 | 2013-04-24 | 中国科学院长春应用化学研究所 | Method for detecting protease activity and inhibitor activity thereof based on chemiluminescence |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3702476B2 (en) * | 1994-08-02 | 2005-10-05 | 日立化成工業株式会社 | Chemiluminescence measurement method |
| JP3711544B2 (en) * | 1994-08-02 | 2005-11-02 | 日立化成工業株式会社 | Chemiluminescence measuring reagent |
| JP3792885B2 (en) * | 1998-03-30 | 2006-07-05 | 大日精化工業株式会社 | Method for measuring peroxidase activity |
-
2015
- 2015-10-12 CN CN201510657942.1A patent/CN105277537B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1756848A (en) * | 2003-04-17 | 2006-04-05 | 坎布里克斯生物科学诺丁汉有限公司 | Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity |
| CN1474185A (en) * | 2003-07-30 | 2004-02-11 | 中国药科大学 | Luminol chemiluminescence immunological analysis detecting method for cardiac muscle calcium protein |
| CN101218354A (en) * | 2005-05-13 | 2008-07-09 | 珀金埃尔默生命与分析科学有限公司 | Use of ammonium and phosphate ions for improving the detection of luciferase |
| CN1687751A (en) * | 2005-05-25 | 2005-10-26 | 段虎 | High stable enhanced chemiluminescence substrate |
| CN1912589A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence measuring method of urea in serum |
| CN101639446A (en) * | 2009-09-07 | 2010-02-03 | 福建省洪诚生物药业有限公司 | Method for detecting blood lactic acid in vitro by using chemiluminescence method |
| CN101639445A (en) * | 2009-09-07 | 2010-02-03 | 福建省洪诚生物药业有限公司 | Method for detecting blood pyruvate in vitro by using chemiluminescence method |
| CN102809541A (en) * | 2012-05-14 | 2012-12-05 | 浙江省海洋开发研究院 | Determination method of enzyme activity of collagenase |
| CN103063657A (en) * | 2012-12-24 | 2013-04-24 | 中国科学院长春应用化学研究所 | Method for detecting protease activity and inhibitor activity thereof based on chemiluminescence |
Non-Patent Citations (8)
| Title |
|---|
| 一种优化的测定过氧化氢酶活性的方法;谭亚芳 等;《生物技术》;20011031;第11卷(第5期);第39-40页 |
| 几种主要化学发光物质的发光性能及其化学发光免疫分析体系;尹东光 等;《标记免疫分析与临床》;20021231;第9卷(第4期);第225-230页 |
| 化学发光法分析测定过氧化氢概况;罗挽兰 等;《咸宁师专学报》;19960831;第16卷(第3期);第30-33页 |
| 化学发光生物传感器法测定食品中有机磷与氨基甲酸酯类农药残留;王晓朋 等;《食品安全质量检测学报》;20141231;第5卷(第12期);第4163-4171页 |
| 化学发光酶免疫分析研究进展;沙栩正 等;《化学发光酶免疫分析研究进展》;20101231;第46卷(第3期);第336-340页 |
| 基于微孔板化学发光法检测碱性磷酸酶的研究;王宇 等;《分析化学研究简报》;20120831;第40卷(第8期);第1279-1283页 |
| 流动注射化学发光法测定过氧化氢酶活性;胡晓捷 等;《浙江大学学报（理学版）》;20031130;第30卷(第6期);第678-680页 |
| 过氧化氢酶活性的化学发光法测定;雷厚成 等;《华西药学杂志》;19941231;第9卷(第3期);第176-178页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105277537A (en) | 2016-01-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TW200510721A (en) | Color reagent, concentration measuring kit, concentration measuring method and sensor chip for use therein | |
| CN102863964B (en) | Manufacturing method of pH ratio fluorescence probe based on organic dye-quantum dot compound | |
| Wang et al. | A low-cost, portable and easy-operated salivary urea sensor for point-of-care application | |
| CN109917134B (en) | Calibrator stabilizer, detection kit for determining C peptide and detection method | |
| ATE386936T1 (en) | SELF-CALIBRATION SYSTEM FOR MAGNETIC BINDING PROCESS | |
| DE60332539D1 (en) | MEMBRANE BASED PROCESS USING TIME RESOLUTION FLUORESCENT MEASUREMENTS | |
| CN104597232A (en) | Captured-antibody competition sandwich immunodetection method capable of extending detection scope and biosensor | |
| CN103951840B (en) | A kind of preparation method and application for detecting salicylic fluorescence sense film | |
| ES2388005T3 (en) | Internal test standardization procedure | |
| CN109470856A (en) | Method of immunity, the system for identifying immunoassays and kit | |
| US9518928B2 (en) | Optode for determining chemical parameters | |
| CN1963512A (en) | Chemiluminescence method for qualitative and quantitative detection of hepatitis B virus | |
| CN106596490B (en) | The supermolecule sensor array and method of synchronous detection paraquat and diquat dibromide | |
| CN105277537B (en) | A method of detection enzymatic activity and chemiluminescence reaction substrate performance | |
| Paul et al. | Calibration of stopped-flow spectrophotometers using a two-step disulfide exchange reaction | |
| RU2007126675A (en) | METHOD FOR DETECTING THE DIFFERENCE BETWEEN THE BLOOD AND THE CONTROL SOLUTION CONTAINING THE SAME ANALYZED MATTER | |
| CN109211883A (en) | The chemiluminometry of concentration of hydrogen peroxide in a kind of detection solution | |
| CN109856094A (en) | A kind of function detecting method of fluorescence immunity analyzer | |
| ATE363069T1 (en) | DETERMINATION OF THE ACCURACY OF A SAMPLE VOLUME IN BIOSENSORS | |
| CN105334189B (en) | Antigen-antibody reaction measuring method based on lenticule imaging | |
| CN108137635A (en) | Acetate complexes and for the quantitative method of acetate | |
| CN108693157B (en) | A method based on 2, 2': preparation and application method of fluorescent chemical sensor of 6 ', 2' -terpyridine derivative | |
| CN106404734B (en) | The method that silver nanoclusters detect potassium concentration infering diing time in vitreous humor | |
| CN110108685A (en) | Using graphene quantum dot as the method for fluorescence probe detection of alkaline phosphatase concentration | |
| CN207096073U (en) | Time-resolved fluorescence enzyme micro-plate reader testing standard plate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: No. 35-305, Changjiang South Road, Xinwu District, Wuxi, Jiangsu 214111 Patentee after: Jiangsu Sanlian Bioengineering Co.,Ltd. Address before: 214000 third floor, zone a, comprehensive incubation building, University Science Park, Xin'an Xingyuan South Road, Wuxi New District, Jiangsu Province Patentee before: SUNLANT BIOLOGICAL ENGINEERING CO.,LTD. |