CN104360074A - Time-resolved fluorescence immunoassay method of Lp-PLA2 and kit - Google Patents
Time-resolved fluorescence immunoassay method of Lp-PLA2 and kit Download PDFInfo
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- CN104360074A CN104360074A CN201410609662.9A CN201410609662A CN104360074A CN 104360074 A CN104360074 A CN 104360074A CN 201410609662 A CN201410609662 A CN 201410609662A CN 104360074 A CN104360074 A CN 104360074A
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- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Abstract
The invention provides a time-resolved fluorescence immunoassay method of Lp-PLA2. The double antibody sandwich method and the time-resolved fluorescence immunoassay method are used together to measure the content of Lp-PLA2. The time-resolved fluorescence immunoassay method comprises the following steps: adding an Lp-PLA2 standard substance or a centrifuged blood serum sample into respective micropores in a microporous plate coated with an Lp-PLA2 resistant monoclonal antibody A; after oscillatory reaction, washing with a concentrated cleaning solution, then adding a biotin labeled Lp-PLA2 resistant monoclonal antibody B; after oscillatory reaction, washing with the concentrated cleaning solution; adding lanthanide labeled streptavidin, and washing with the concentrated cleaning solution after oscillatory reaction; after adding an enhancement solution and oscillating, performing fluorescence emitting under excitation of ultraviolet light, testing the fluorescence strength cps with a time-resolved fluorescence spectrometer, and determining the content of LP-PLA2 in a sample to be tested in comparison with a standard curve. The time-resolved fluorescence immunoassay method can quantitatively test the content of Lp-PLA2 in the sample, and has the characteristics of high sensitivity, wide analysis range, good stability, quick detection and easy automation.
Description
Technical field
The present invention relates to technical field of biological, relate to the time-resolved fluoroimmunoassay detection method of plasma lipoprotein phospholipase A2 (Lp-PLA2) particularly.
Background technology
Lp-PLA2 belongs to phospholipase A2 superfamily, secretes primarily of inflammatory cell (as macrophage, lymphocyte etc.).The protein that Lp-PLA2 is made up of 441 amino acid, relative molecular weight is about 45400, with all the other members of phospholipase A2 family unlike, Lp-PLA2 does not need calcium ion to maintain its catalytic activity.Lp-PLA2 has the activity of degraded platelet activating factor, and be therefore also referred to as platelet-activating factor acetylhydrolase, it exists in a large number at atherosclerotic position, and can be regulated by inflammatory mediator, has the effect of pro-atherogenic.Lp-PLA2 is a kind of new Inflammatory parameter, is the specific marker thing of reflection vascular inflammation.
Enzyme linked immunosorbent assay, chemiluminescence immunoassay, latex-enhanced turbidimetry and colloidal gold method principle product is mainly at present for the reagent detecting Lp-PLA2.But it is long to there is detection time in euzymelinked immunosorbent assay (ELISA), poor repeatability, the not high defect of automaticity.Chemiluminescence immunoassay existence and stability and poor repeatability, the defects such as testing cost is high.The defects such as it is narrow that latex-enhanced turbidimetry has the range of linearity, poor sensitivity.And also there is the weak defect of sensitivity in colloidal gold method.
Timed resolved fluoroimmunoassay is with lanthanide series labelled antigen or antibody, and the novel on-radiation trace analysis of one combining with time resolution determination techniques and set up, there is the features such as highly sensitive, stable luminescence, fluorescence lifetime long, natural fluorescence disturbs less, standard curve range is wide.Widespread use in clinical detection at present.Also do not report the immunologic detection method being specifically designed to the Time-resolved fluorescence assay detecting Lp-PLA2 at present, do not have corresponding kit to provide yet.
Summary of the invention
Goal of the invention: for solving problems of the prior art, the invention provides a kind of time-resolved fluoroimmunoassay detection method setting up a kind of plasma lipoprotein phospholipase A2 based on Time-resolved fluorescence assay, the clinical analysis for Lp-PLA2 provide a kind of highly sensitive, analyst coverage is wide, good stability, detection fast and be easy to the detection technique of robotization.
Technical scheme: for realizing above-mentioned technical purpose, the present invention establishes a kind of time-resolved fluoroimmunoassay detection method of plasma lipoprotein phospholipase A2, it is characterized in that, double antibody sandwich method binding time resolved fluorometric immunologic detection method is adopted to measure the content of plasma lipoprotein phospholipase A2 (Lp-PLA2), comprise the steps: on the microwell plate being coated with anti-Lp-PLA2 monoclonal antibody A, add the serum sample of Lp-PLA2 standard items or centrifugal rear acquisition in respective micropore, wash with cleansing solution after oscillating reactions, then biotin labeled anti-Lp-PLA2 monoclonal antibody B is added, cleansing solution washing after oscillating reactions, add the Streptavidin of lanthanide series mark, then wash with cleansing solution after oscillating reactions, add after strengthening liquid vibration, emitting fluorescence under the exciting of ultraviolet light, measures its fluorescence intensity cps with time-resolved fluorescence instrument, and reference standard curve can determine Lp-PLA2 content in testing sample.
Wherein, described lanthanide series is selected from Eu
3+, Tb
3+, Sm
3+, Nd
3+and Dy
3+in any one.Preferably, described lanthanide series is Eu
3+.
Particularly, the Lp-PLA2 monoclonal antibody A be coated on microwell plate identifies the different epi-position of Lp-PLA2 respectively from biotin labeled anti-Lp-PLA2 monoclonal antibody B and is independent of each other, wherein, Lp-PLA2 monoclonal antibody A, B are prepared by monoclonal hybridoma technology or are obtained by phage antibody technology screening.
Described cleansing solution is the 0.01M phosphate buffer (1 × PBST) containing 0.05wt%TWEEN-20.
Described enhancing liquid comprises following component: the Potassium Hydrogen Phthalate damping fluid of 1L pH 3.2 is containing 15 μm of ol β-naphthalene formyl trichloroacetones, 50 μm of ol trioctyl-phosphine oxide TOPO and 1ML triton x-100s.
Method of the present invention can replace with solid phase competition law or solid phase antigen competition law equally.Particularly, when using insolubilized antibody competition law, by Lp-PLA2 and Eu
3+labelled antigen and insolubilized antibody generation competition binding, add fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
When using solid phase antigen competition law, by Lp-PLA2 and the quantitative Eu of solid phase antigen competitive binding
3+labelled antibody, adds fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
The present invention proposes a kind of time-resolved fluoroimmunoassay detection kit of plasma lipoprotein phospholipase A2 (Lp-PLA2) simultaneously, it comprises as lower part: be coated with the microwell plate of anti-Lp-PLA2 monoclonal antibody A, concentrated washing lotion, strengthen liquid, the Streptavidin of lanthanide series mark and biotin labeled anti-Lp-PLA2 monoclonal antibody B, wherein, the described microwell plate being coated with anti-Lp-PLA2 monoclonal antibody A and preparing by the following method with biotin labeled Lp-PLA2 monoclonal antibody B:
(1) preparation of Lp-PLA2 monoclonal antibody A and Lp-PLA2 monoclonal antibody B and purifying:
With increment cultivation amplified hybridization oncocyte, gained cell liquid carries out purifying through sad-saturated ammonium sulfate method, obtains monoclonal antibody.
(2) the bag quilt of microwell plate:
Lp-PLA2 monoclonal antibody A coating buffer after purifying is diluted to 1-10ng/ml, the 96 each holes of microwell plate add 100ul, and 4 DEG C of placements are spent the night; Discard coating buffer, rinse twice with the 0.01M phosphate buffer containing 0.05wt%TWEEN-20, add 150ul confining liquid and close, 4 DEG C of placements are spent the night.Discard confining liquid, vacuum is drained, and places-20 DEG C of freezen protective after lath sealing;
(3) biotin labeling of Lp-PLA2 monoclonal antibody B: use phosphate buffer to be diluted to 1-10ng/ml Lp-PLA2 monoclonal antibody B after purifying, then the biotin that Lp-PLA2 monoclonal antibody B and NHS activates is pressed 1:1 mixing, lucifuge reaction 30min, remove the NSH-Biotin that may have neither part nor lot in mark with the ultra filtration membrane post of 50kDa, obtain biotin labeled Lp-PLA2 monoclonal antibody B.
Wherein, the Lp-PLA2 monoclonal antibody A be coated on microwell plate identifies the different epi-position of Lp-PLA2 respectively from biotin labeled anti-Lp-PLA2 monoclonal antibody B and is independent of each other, wherein, Lp-PLA2 monoclonal antibody A, B are prepared by monoclonal hybridoma technology or are obtained by phage antibody technology screening.
Particularly, described coating buffer is pH9.6,50mmol/L Na
2cO
3-NaHCO
3damping fluid, described confining liquid is the phosphate buffer containing 0.05wt%TWEEN-20,3g/L BSA.
Described concentrated washing lotion is the 0.2M phosphate buffer (20 × PBST) containing 1wt%TWEEN-20.
Described lanthanide series is selected from Eu
3+, Tb
3+, Sm
3+, Nd
3+and Dy
3+in any one.Preferably, described lanthanide series is Eu
3+.
Described enhancing liquid comprises following component: the Potassium Hydrogen Phthalate damping fluid of 1L pH 3.2 is containing 15 μm of ol β-naphthalene formyl trichloroacetones, 50 μm of ol trioctyl-phosphine oxide TOPO and 1ML triton x-100s.
Beneficial effect: compared with prior art, tool of the present invention has the following advantages:
(1) the clinical cut-off value of Lp-PLA2 is 175ng/ml, is included in the sensing range of this kit, can meets clinical needs;
(2) analyst coverage is wide, sensing range 50-3000ng/ml;
(3) measure fast, be easy to robotization;
(4) lanthanide series is used to mark, no radioactivity pollute
(5) antijamming capability is strong, according to the luminous characteristics of lanthanide chelate, effectively can get rid of the interference of non-specific fluorescence.
Accompanying drawing explanation
The Time-resolved fluorescence assay examination criteria curve map of Fig. 1: Lp-PLA2;
Fig. 2: Lp-PLA2 Time-resolved fluorescence assay method detection reaction schematic diagram.Wherein, A:Lp-PLA2 monoclonal antibody; B:Lp-PLA2 albumen; C: biotin labeled anti-Lp-PLA2 monoclonal antibody; D:Eu
3+-Streptavidin compound.
Embodiment
Embodiment is further illustrating Lp-PLA2 Time-resolved fluorescence assay detection method provided by the present invention, but working of an invention is not limited thereto, and the replacement that any function is identical all belongs to protection scope of the present invention.
The preparation of embodiment 1Lp-PLA2 Time-resolved fluorescence assay detection kit.
(1) preparation of experimental solutions:
Concentrated washing lotion: containing the 0.2M phosphate buffer (20 × PBST) of 1wt%TWEEN-20;
Cleansing solution: containing the 0.01M phosphate buffer of 0.05wt%TWEEN-20; This cleansing solution obtains by concentrating washing lotion dilution;
Strengthen liquid: 1L pH3.2 Potassium Hydrogen Phthalate damping fluid is containing 15 μm of ol β-naphthalene formyl trichloroacetones (β-NTA), 50 μm of ol trioctyl-phosphine oxide TOPO and 1mL triton x-100s (Triton X100).
(2) preparation and purification of the preparation of Lp-PLA2 monoclonal antibody A and Lp-PLA2 monoclonal antibody B:
Lp-PLA2 monoclonal antibody A and Lp-PLA2 monoclonal antibody B be identify respectively Lp-PLA2 different epi-position and be independent of each other, wherein, Lp-PLA2 monoclonal antibody A and B all can be prepared by monoclonal hybridoma technology or be obtained by phage antibody technology screening.Directly adopting in the present embodiment is two kinds of monoclonal antibodies that are commercially available, that identify the different epi-position of Lp-PLA2.Wherein, Lp-PLA2 monoclonal antibody A selects anti-human lipoprotein phospholipase A2 (Lp-PLA 2) monoclonal antibody (article No. CLM0019006, buy from constant engineering in medicine company limited of Shenzhen), Lp-PLA2 monoclonal antibody B selects anti-human lipoprotein phospholipase A2 (Lp-PLA 2) monoclonal antibody (article No. CLM0019005, buy from constant engineering in medicine company limited of Shenzhen), two kinds of monoclonal antibodies identify the different epi-positions of Lp-PLA2 respectively.Certainly, two kinds of monoclonal antibodies can exchange use, as long as ensure the different epi-positions of two kinds of monoclonal antibody identification Lp-PLA2.
Respectively with increment cultivation amplified hybridization oncocyte, gained cell liquid carries out purifying through sad-saturated ammonium sulfate method, obtains monoclonal antibody.
(3) bag is by the preparation of plate:
By Lp-PLA2 monoclonal antibody A 50mmol/L Na after purifying
2cO
3-NaHCO
3pH9.6 damping fluid is diluted to 8ng/ml, and the 96 each holes of microwell plate add 100ul, and 4 DEG C of placements are spent the night.Discard coating buffer, rinse twice with cleansing solution (phosphate buffer (1 × PBST) containing 0.05%TWEEN-20), add the above-mentioned buffer blind of 150ul containing 3g/LBSA, 4 DEG C of placements are spent the night.Discard confining liquid, vacuum is drained, and places-20 DEG C of freezen protective after lath sealing.
(5) antibody biotin mark:
The biotin labeling of Lp-PLA2 monoclonal antibody B: use phosphate buffer to be diluted to 8ng/ml Lp-PLA2 monoclonal antibody B after purifying, then the biotin that Lp-PLA2 monoclonal antibody B and NHS activates is pressed 1:1 mixing, lucifuge reaction 30min, removes the NSH-Biotin that may have neither part nor lot in mark with the ultra filtration membrane post of 50kDa.
(5) test:
A: the preparation of sample: gather patient blood sample, centrifugal 5 minutes of 1500r/min
B: detect:
Double antibody sandwich method: the microwell plate lath getting coated antibody, add the serum sample of 100 μ l positive quality control product Lp-PLA2 standard items or centrifugal rear acquisition in respective micropore, negative control hole PBS replaces.After 37 DEG C of vibration 1h, cleansing solution washes 3 times.Every hole adds the biotin labeled Lp-PLA2 monoclonal antibody of 100ul, after 37 DEG C of vibration 1h, washs liquid and washes 3 times.Every hole adds 100ul Eu
3+-Streptavidin compound, after 37 DEG C of vibration 1h, cleansing solution washes 3 times.Every hole adds 200ul and strengthens liquid, and 37 DEG C of vibration 15min, with time resolution analyser reading.From the Lp-PLA2 content typical curve calculation sample, see Fig. 1.
Should be noted that before the assay: before use, all reagent is gone up to room temperature (18-30 DEG C); Immediately all reagent is put back to 2-8 DEG C after use; If the large suggestion of sample size uses Multi-channel liquid transfer device; In all constant-temperature incubation processes, avoid light to irradiate, use cap covers micropore.
Possible alternative:
Insolubilized antibody competition law: by Lp-PLA2 and Eu
3+labelled antigen and insolubilized antibody generation competition binding, add fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
Solid phase antigen competition law: by Lp-PLA2 and the quantitative Eu of solid phase antigen competitive binding
3+labelled antibody, adds fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
The degree of accuracy of embodiment 2Lp-PLA2 Time-resolved fluorescence assay detection kit, accuracy and stability test.
1. kit accuracy, Accuracy Analysis experiment:
Carry out 20 times to the standard solution (phosphate buffer of pH7.4,0.05M) of 207ng/ml Lp-PLA2 to detect.207ng/ml is the critical value that Lp-PLA2 predicts cardiovascular and cerebrovascular disease high risk, and the pinpoint accuracy of method is conducive to the accurate judgement to onset risk.The mean value of 20 testing results is 204.64ng/ml, and standard deviation is 8.26, and aberration rate is 4.03%.Result shows, and testing result and actual concentrations are comparatively close, and accuracy is higher, and aberration rate <5%, has good degree of accuracy simultaneously.
2. stabilization of kit test:
Kit preservation condition is 2-8 DEG C, preserves after 6 months, measures the indices of kit, finds all within normal range.Consider in transport and use procedure, have improper preservation condition and occur, placed 6 days under 37 DEG C of conditions of preserving by kit, carry out accelerated aging tests, result shows that this kit indices meets the requirements completely.Consider that the freezing situation of kit occurs, kit is put into-20 DEG C of refrigerator freezings 5 days, measurement result also shows that kit indices is completely normal.Can show that kit at least can preserve more than 6 months at 2-8 DEG C from above result.
Embodiment 3Lp-PLA2 Time-resolved fluorescence assay detection kit compares with ELISA detection kit.
Select the Lp-PLA2 standard solution of high, medium and low three concentration, analyze with this kit and ELISA detection kit simultaneously, each Concentration Testing 5 times, compares acquired results, determines the consistance of two kinds of methods.High concentration selects 1000ng/ml, and middle concentration selects 300ng/ml, and low concentration selects 80ng/ml.The testing result obtained is as following table:
| Average 1 (height) | S 1 | Average 2 (in) | S 2 | Average 3 (low) | S 3 | |
| ELISA | 995.56 | 45.67 | 304.34 | 14.76 | 78.35 | 3.4 |
| TRIFA | 1002.45 | 48.65 | 297.36 | 13.67 | 78.12 | 3.87 |
Two groups of data are done to the t inspection of paired sample, obtain P>0.05, both difference not statistically significants, illustrates two kinds of methods no matter height, in or low concentration Lp-PLA2 inspection in, all have good consistance.
Claims (10)
1. the time-resolved fluoroimmunoassay detection method of a plasma lipoprotein phospholipase A2, it is characterized in that, double antibody sandwich method binding time resolved fluorometric immunologic detection method is adopted to measure the content of plasma lipoprotein phospholipase A2 (Lp-PLA2), comprise the steps: on the microwell plate being coated with anti-Lp-PLA2 monoclonal antibody A, add the serum sample of Lp-PLA2 standard items or centrifugal rear acquisition in respective micropore, wash with cleansing solution after oscillating reactions, then biotin labeled anti-Lp-PLA2 monoclonal antibody B is added, cleansing solution washing after oscillating reactions; Add the Streptavidin of lanthanide series mark, then wash with cleansing solution after oscillating reactions; Add after strengthening liquid vibration, emitting fluorescence under the exciting of ultraviolet light, measures its fluorescence intensity cps with time-resolved fluorescence instrument, and reference standard curve can determine Lp-PLA2 content in testing sample.
2. detection method according to claim 1, is characterized in that, described lanthanide series is selected from Eu
3+, Tb
3+, Sm
3+, Nd
3+and Dy
3+in any one.
3. detection method according to claim 1, it is characterized in that, the Lp-PLA2 monoclonal antibody A be coated on microwell plate identifies the different epi-position of Lp-PLA2 respectively from biotin labeled anti-Lp-PLA2 monoclonal antibody B and is independent of each other, wherein, Lp-PLA2 monoclonal antibody A, B are prepared by monoclonal hybridoma technology or are obtained by phage antibody technology screening.
4. detection method according to claim 1, is characterized in that, described cleansing solution is the 0.01M phosphate buffer containing 0.05wt%TWEEN-20.
5. detection method according to claim 1, it is characterized in that, described enhancing liquid comprises following component: the Potassium Hydrogen Phthalate damping fluid of 1L pH 3.2 is containing 15 μm of ol β-naphthalene formyl trichloroacetones, 50 μm of ol trioctyl-phosphine oxide TOPO and 1ML triton x-100s.
6. method according to claim 1, is characterized in that, described double antibody sandwich method can substitute with insolubilized antibody competition law or solid phase antigen competition law:
Wherein, when replacing with insolubilized antibody competition law, by Lp-PLA2 and Eu
3+labelled antigen and insolubilized antibody generation competition binding, add fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
When replacing with solid phase antigen competition law, by Lp-PLA2 and the quantitative Eu of solid phase antigen competitive binding
3+labelled antibody, adds fluorescence enhancement solution after inculation washing in solid phase, and measure fluorescence intensity, measured fluorescence intensity and Lp-PLA2 content are negative correlation.
7. the time-resolved fluoroimmunoassay detection kit of a plasma lipoprotein phospholipase A2 (Lp-PLA2), it is characterized in that, comprise as lower part: the microwell plate being coated with anti-Lp-PLA2 monoclonal antibody A, concentrated washing lotion, strengthen liquid, the Streptavidin of lanthanide series mark and biotin labeled anti-Lp-PLA2 monoclonal antibody B, wherein, the Lp-PLA2 monoclonal antibody A be coated on microwell plate identifies the different epi-position of Lp-PLA2 respectively from biotin labeled anti-Lp-PLA2 monoclonal antibody B and is independent of each other, the described microwell plate being coated with anti-Lp-PLA2 monoclonal antibody A and preparing by the following method with biotin labeled Lp-PLA2 monoclonal antibody B:
(1) preparation of Lp-PLA2 monoclonal antibody A and Lp-PLA2 monoclonal antibody B and purifying:
With increment cultivation amplified hybridization oncocyte, gained cell liquid carries out purifying through sad-saturated ammonium sulfate method, obtains Lp-PLA2 monoclonal antibody A and Lp-PLA2 monoclonal antibody B respectively;
(2) the bag quilt of microwell plate:
Lp-PLA2 monoclonal antibody A coating buffer after purifying is diluted to 1 ~ 10ng/ml, and the 96 each holes of microwell plate add 100ul, and 4 DEG C of placements are spent the night; Discard coating buffer, rinse twice with the phosphate buffer containing 0.05wt%TWEEN-20, add 150ul confining liquid and close, 4 DEG C of placements are spent the night.Discard confining liquid, vacuum is drained, and places-20 DEG C of freezen protective after lath sealing;
(3) biotin labeling of Lp-PLA2 monoclonal antibody B: use phosphate buffer to be diluted to 1 ~ 10ng/ml Lp-PLA2 monoclonal antibody B after purifying, then the biotin that Lp-PLA2 monoclonal antibody B and NHS activates is pressed 1:1 mixing, lucifuge reaction 30min, remove the NSH-Biotin that may have neither part nor lot in mark with the ultra filtration membrane post of 50kDa, obtain biotin labeled Lp-PLA2 monoclonal antibody B.
8. kit according to claim 7, is characterized in that, described coating buffer is pH9.6,50mmol/L Na
2cO
3-NaHCO
3damping fluid, described confining liquid is the phosphate buffer containing 0.05wt%TWEEN-20,3g/LBSA.
9. kit according to claim 7, is characterized in that, described concentrated washing lotion is the 0.2M phosphate buffer containing 1wt%TWEEN-20.
10. kit according to claim 7, it is characterized in that, described enhancing liquid comprises following component: the Potassium Hydrogen Phthalate damping fluid of 1L pH 3.2 is containing 15 μm of ol β-naphthalene formyl trichloroacetones, 50 μm of ol trioctyl-phosphine oxide TOPO and 1ML triton x-100s.
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|---|---|---|---|
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| CN104820097A (en) * | 2015-05-22 | 2015-08-05 | 北京协和洛克生物技术有限责任公司 | Liquid-chip kit for quantitatively detecting concentration of lipoprotein phospholipase A2 in sample and preparation method thereof |
| CN104849456A (en) * | 2015-05-18 | 2015-08-19 | 北京协和洛克生物技术有限责任公司 | Reagent kit for detecting concentration of Lp-PLA2 and preparing method thereof |
| CN108152500A (en) * | 2017-12-27 | 2018-06-12 | 江南大学 | A kind of Microcystin immune quantitative test paper item based on fluorescent microsphere label |
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Application publication date: 20150218 |