CN107064071A - Determine rutin and the surface plasma body resonant vibration analysis method of protein binding level - Google Patents
Determine rutin and the surface plasma body resonant vibration analysis method of protein binding level Download PDFInfo
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Abstract
Rutin and surface plasma body resonant vibration (SPR) analysis method of protein binding level are determined the invention discloses a kind of, analysis object is the Interaction System of rutin and albumen, and analysis target is the equilibrium constant for trying to achieve interaction reaction.Rutin pyridinium dissolution and diluted in this method with the cushioning liquid containing pyridine, the problem of solving rutin solubility is not enough in aqueous, be allowed to produce sufficiently strong spr signal.In addition, the dilute solution of same composition but the pyridine without rutin has been prepared during analysis correspondingly in the rutin solution of prepared and diluted, for determining influence of the pyridine to spr signal, accurately to obtain the interaction signal of rutin generation.
Description
Technical field
The present invention relates to the application process of Applications of surface plasmon resonance in analytical chemistry, and in particular to one kind is used to survey
Determine the surface plasma body resonant vibration analysis method of rutin and protein interaction combination level.
Background technology
Surface plasma body resonant vibration (surface plasmon resonance, SPR) technology is a kind of intermolecular mutual
Widely used method in the analysis field of effect.Its general principle is to utilize incident light irradiation SPR sensorgram chip so that incident
The evanescent wave that light is produced resonates with censorchip surface metallic surface plasma wave, influences the property of emergent light；Hereafter
Occur interaction reaction in censorchip surface, cause the change of resonance condition, cause outgoing light property to change, to it
It is monitored the detection that Thermodynamic parameters reaction can be achieved.During general SPR analyses, one of interaction reaction
Participate in thing and be fixed in SPR detection chips surface often through covalent bond, electrostatic interaction, hydrophobic effect etc. mode, then will
The detection chip surface is flowed through after the participation thing dissolving of another (or several) reaction.When occurring interaction reaction,
The combination or dissociation of reactant change the surface nature for causing detection chip, are reflected as the change of instrument signal.SPR skills
Art can carry out quick, sensitive, unmarked analysis, be often used in the speed constant for determining interaction reaction, balance normal
The physical-chemical datas such as number.
Rutin (rutin) is a kind of flavone compound, widely distributed in nature, particularly plant, in some plants
It can be found in the organs such as root, stem, leaf, the fruit of thing and tissue.The structural formula of rutin is as follows：
Rutin has obvious physiologically active, is common food and Chinese medicine, such as buckwheat, ginkgo, the sophora bud, matrimony vine, rue
Important activity composition in perfume, motherwort, aloe, radix bupleuri, folium eucalypti, and tobacco.The structural formula of rutin is as shown in Figure 1.Rutin
With stronger antioxidation activity, can be maintained in human body the tension force of vascular wall, improve its elasticity, reduction vascular wall fragility and
Permeability, thus microcirculation, reduction blood fat and cholesterol level can be improved, can be in the treatment of cardiovascular and cerebrovascular disease
Play a role.In addition, rutin also has certain antibacterial and anti-aging effects, and the secretion and raising of insulin can be influenceed
Insulin receptor affinity, also there is certain application in treating diabetes field.Rutin enter human body after, it is necessary to by with it is many
Plant protein and occur interaction to realize its physiology, pharmacological function, such as needed when its concentration is higher and the human blood in blood
Pure albumen (human serum albumin, HSA) occur to combine so as to be transported to blood circulation other organs or
Tissue, is then dissociated to play physiological function again, and its efficacy exertion is also with it with biology enzyme or special and non-specific acceptor
With reference to relevant.Therefore, research rutin process relevant with protein interaction, particularly tries to achieve it and occurs with multiple proteins
The equilibrium constant of association reaction, studies significant to its pharmacology, pharmacokinetics etc..
As quick, unmarked, the real-time analysis means of one kind, SPR methods are substantially suitable for rutin-protein phase interaction
Analysis, but there is in implementation process the solubility problem of difficulty, i.e. rutin.SPR is a kind of mass sensitive
Analysis method, its signal intensity depends on the size of (in 50 nanometers) measured object quality near censorchip surface, therefore works as quilt
When analyte is the small-molecule substances such as rutin, it must have sufficiently large solubility in water environment, to form sufficiently strong letter
Number.Especially, in order to simulate physiological environment, SPR analyses process often uses the cushioning liquid containing inorganic salts to prepare sample
Solution, this further causes the problem of rutin solubility is reduced.Had no in existing patented method using SPR method research rutins and
The method of protein interaction.Therefore, it is necessary to the dissolving of rutin in aqueous that improves for passing through analytical procedure
Property, and the fast analysis determination method of the rutin based on SPR-protein interaction combination level is developed based on this.
The content of the invention
In order to make up analysis difficulty caused by the deficiency of existing method, particularly rutin solubility, mesh of the invention
Be to provide a kind of new interaction analyzing method based on Surface Plasmon Resonance Technology, realize to rutin and protein
Interaction combines the measure of level (i.e. the equilibrium constant of association reaction).
The analysis method of the present invention includes the cleaning of sensing chip, the carboxylated modification of chip surface, protein solution pH
Screening, the activation of chip surface carboxyl, the covalent bond of protein, the dissolving of rutin and dilution, protein and rutin it is mutual
The steps such as measure, the data analysis of mechanism.Detailed process is as follows, and each step must be carried out successively.
1st, the cleaning of sensing chip.
When SPR sensorgram chip (being usually gold-plated sheet glass) is Demountable, sensing chip is integrally immersed in
By concentrated ammonia liquor, 30% hydrogen peroxide, water by volume 1:1:In 5 solution mixed, it is placed in 60-95 DEG C of environment at least
10 minutes, then cleaned, used after drying with water.When sensing chip is that (i.e. metal level is directly plated on rib to non-dismountable formula structure
Mirror surface) when, above-mentioned solution is added dropwise in chip surface detection window position, is placed in less than 90 DEG C but as high as possible (does not damage
The structures such as prism) at a temperature of at least 30 minutes, surface is then cleaned with water and used after drying.Brand-new sensing can also be used
Chip.
It is preferred that, each analysis process uses brand-new sensing chip, to be coated with the sheet glass of exposed golden film, disposably makes
With being discarded after the completion of analysis.
2nd, the carboxylated modification of chip surface.
The sensing chip of cleaning is installed after SPR instruments, the solution for meeting following condition is flowed through chip surface：
Solute is the straight-chain carboxylic acid that a kind of sulfydryl replaces, and wherein sulfydryl is located at another end of carbochain away from carboxyl；Carbon chain lengths are
Containing three to 20 carbon atoms, i.e. mercaptopropionic acid, mercaptobutyric acid until one of sulfydryl arachic acid；Solute concentration is 2mmol/L
To 50mmol/L；Solvent is water and/or ethanol, depending on concentration needed for solute can be dissolved to by whichever.In the process of circulation in real time
Monitor spr signal, the currency, but at least 1 minute, solution flow rate was after sample introduction untill spr signal no longer changes
0.1 mul/min to 1 ml/min, depending on flow cell volume.Whole pipeline and core are rinsed with water after the completion of above step
Piece surface.Chip area temperature is 0 DEG C to 60 DEG C during circulation.
It is preferred that, using the 5mmol/L 3- mercaptopropionic acids aqueous solution with 5 mul/min of flow rate 30 minutes, chip
37 DEG C of controlling temperature with region, after the completion of with water flushing line and chip surface.
3rd, the pH screenings of protein solution.
The targeted target protein of this method is water soluble protein, and its isoelectric point is more than 4.Monitoring spr signal, makes in real time
Chip surface is flowed through with the solution for meeting following condition：Protein concentration is 1 nanograms/milliliter into 50 mg/mls
Some value；Solvent is acetic acid-sodium acetate buffer solution, and Na ion concentration is a value no more than 50mmol/L；PH is 2.5
At least three value into 6.0, scope must cover at least two pH units, and interval is at least 0.2, is at most 1 pH unit.That is,
The solution of circulation is several ankyrin species solution different with concentration, fixed buffer concentration but pH.Each solution circulation
The identical time, between 5 seconds and 60 minutes, flow velocity is identical, between 0.1 mul/min between 1 ml/min.Chip
Regional temperature is a value between 0 DEG C to 60 DEG C.The SPR when protein solution for comparing each different pH flows through the chip of carboxyl modified
The change of signal.If in the several pH value attempted, under the used currency, each solution makes spr signal reach surely
It is fixed, then select signal after stabilization to differ the pH of maximum solution with baseline as subsequent analysis；If under the currency used
Each solution does not make signal reach stabilization, then selection signal changes the pH of most fast solution as subsequent analysis；If part reaches
Steady component is not up to stabilization, then selection circulation completes time-ofday signals and the pH of maximum solution is differed with baseline as subsequent analysis.
After the completion of this step whole pipeline and chip are rinsed with water.
It is preferred that, protein is dissolved separately in Na ion concentration for 10mmol/L, pH is respectively 4.0,4.5,5.0,5.5
Acetic acid-sodium acetate buffer solution in, with 5 mul/min of flow velocity respectively circulation 5 minutes, 37 DEG C of temperature control, signal at the end of selection
Change the pH of maximum solution.
4th, the activation of chip surface carboxyl.
The carboxyl for the mercaptan carboxylic acid that one of following two methods modify chip surface is selected to be activated.
First, making the solution for meeting following condition flow through chip surface：Solute is EDC, i.e. N- (3- dimethylaminos third
Base)-N '-ethyl-carbodiimide hydrochloride and NHS, i.e. N- hydroxysuccinimides, both concentration are 0.0001 to 0.5 gram/
Milliliter, and EDC concentration is not less than NHS concentration, flow velocity between 0.1 mul/min between 1 ml/min, the currency with
Spr signal, which no longer changes, to be defined, but at least 1 minute.Chip area temperature is a value between 0 DEG C to 60 DEG C.
Second, successively making EDC solution and NHS solution flow through chip surface respectively, respective concentration range, flow velocity are same
On, the currency is no longer changed by respective spr signal to be defined, but is each at least 1 minute.Chip area temperature is 0
DEG C to a value between 60 DEG C.
It is preferred that, circulated 30 minutes with EDC and NHS mixed solution, both concentration are respectively 0.01 and 0.002 gram/milli
Rise, flow velocity is 5 mul/min.Temperature control is 37 DEG C.
5th, the covalent bond of protein.
By target protein with the buffer solution determined in the 3rd step, protein concentration be 1 nanograms/milliliter to 50 milligrams/
Between milliliter, then it is allowed to flow through chip surface, chip area temperature is a value between 0 DEG C to 60 DEG C.Flow velocity between
0.1 mul/min is defined between 1 ml/min, the currency is no longer changed by spr signal, but at least 1 minute.Complete
Afterwards with water flushing line and chip.
It is preferred that, using 0.01 grams per milliliter, the protein solution that pH is 5.0, with 5 mul/min of flow rates 20 minutes,
37 DEG C of temperature control.
6th, the dissolving and dilution of rutin.
Select phosphate buffer solution (PBS), Tris cushioning liquid (TRIS buffer, TBS) or
One of HEPES cushioning liquid (N-2- hydroxyethyl piperazine -2 '-ethanesulfonic acid buffers of-N ', HBS) solution, pH is 6.6 at 25 DEG C
To 8.5,3% to 15% pyridine (volume fraction) is added thereto.Hereafter other appropriate additives, such as ethylenediamine tetrem are added
Sour sodium (EDTA) and surfactant P20 etc., total mass fraction are no more than 5%.The retarder thinner of rutin is used as using this solvent.
Rutin is dissolved in pure pyridine, concentration is a value in 1-50mmol/L.This reed is diluted with above-mentioned retarder thinner
The pyridine solution of fourth, at least obtains at least differing 20% between 3 various concentrations, and each concentration.Each dilution water contains rutin
Pyridine when, with same ratio dilute a undissolved rutin pure pyridine, obtain the pyridine solution of various concentrations, its pyridine contains
Amount and the solution containing rutin of above-mentioned dilution are same.Rutin concentration is designated as c respectively in each solution1、c2、…ci。
It is preferred that, 10mmol/LHEPES, 0.15mol/L sodium chloride, 3mmol/L (are contained with HBS-EP+ buffer solutions
EDTA, the surfactant P20 of 0.005% volume fraction, pH is 7.4) 8% (volume fraction) pyridine of middle addition at 25 DEG C
Solution is diluent, and rutin is dissolved in into pure pyridine to 10mmol/L, be then diluted to 0.06 with this diluent, 0.10,0.20,
0.25th, 0.30mmol/L, dilutes the pure pyridine solution for being free of rutin during dilution with same ratio every time.
7th, the measure of protein and rutin interaction process.
Circulation solution in SPR instrument pipelines is replaced with to rutin retarder thinner used in the 6th step.It will be obtained in 6th step
After the dilution of the rutin of each concentration arrived solution with it is corresponding respectively without rutin dilution pyridine solution according to by it is dilute to dense,
The order of rutin is sequentially sent to SPR instruments and analyzed after first pyridine.Sample flow rate is between 0.1 mul/min to 1 ml/min
Between clock, between 0 DEG C to 60 DEG C of chip area temperature control.Tracer signal is until no longer change, and at least maintain 1 after each sample introduction
Minute.Each sample introduction is until after the close stabilization of signal, stop sample introduction, with the dodecyl sulphur between mass fraction 1% to 10%
Sour sodium (SDS) solution flushing line and chip are until hereafter solution in pipeline is replaced with institute in the 6th step by signal again close to stable
Retarder thinner, is analyzed next time.Each concentration is recorded for ciRutin solution sample introduction and signal stabilization after letter
Number value, be designated as Ri, each and ciSame dilution, but the signal value after the solution sample introduction of rutin and signal stabilization is free from for ri。
It is preferred that, 37 DEG C of temperature control, sample flow rate is 5 mul/min during analysis every time, and the SDS mass fractions used are
5%.
8th, data analysis.
Protein used is tried to achieve according to following steps, and the equilibrium constant of association reaction occurs with rutin.
A, as described in step 7, record each ciWhen RiAnd ri；
B, calculate each 1/ciWith 1/ (Ri-ri), and with 1/ (Ri-ri) to 1/ciMapping, 1/ciFor abscissa, 1/ (Ri-ri)
For ordinate.
C, linear fit is carried out to data point in figure, the intercept of gained equation is designated as b, and slope is designated as a, then association reaction
The equilibrium constant is b/a, the inverse of unit concentration unit used in experiment.
In the analysis method of the present invention, rutin pyridinium dissolution is simultaneously diluted with the cushioning liquid containing pyridine, to solve reed
Fourth is allowed to produce sufficiently strong spr signal the problem of solubility is not enough in aqueous.In addition, preparing dilute during analysis
The dilute solution of same composition but the pyridine without rutin is prepared during the rutin solution released correspondingly, for determining pyridine
Influence to spr signal, accurately to obtain the interaction signal of rutin generation.
Brief description of the drawings
Fig. 1：The present invention determines rutin using Applications of surface plasmon resonance and the analysis process of protein binding level is shown
It is intended to.
Fig. 2：7th step in embodiments of the invention, HSA protein interacts with solution after each dilution containing rutin
SPR testing result figures, for trying to achieve each Ri。
Fig. 3：7th step in embodiments of the invention, HSA protein and each pyridine dilute solution phase interaction for not containing rutin
SPR testing result figures, for trying to achieve each ri。
Fig. 4：8th step in embodiments of the invention, maps and linear fit result.
Embodiment
Below in conjunction with accompanying drawing, by embodiment, technical scheme is expanded on further, but the protection model of the application
Enclose and do not limited by the actual conditions of these embodiments.
Embodiment：Determine rutin using the analysis method in the present invention and interacted with human serum albumins (HSA) and react
The equilibrium constant.Comprise the following steps that.
1st, using brand-new sensing chip, to be coated with the sheet glass of exposed golden film.It is installed into SPR instruments, starts instrument
Device, chip area temperature control maintains 37 DEG C.
2nd, using the 5mmol/L 3- mercaptopropionic acids aqueous solution with 5 mul/min of flow rate 30 minutes, chip area
37 DEG C of temperature control, after the completion of with water flushing line and chip surface.
3rd, HSA is dissolved in Na ion concentration for 10mmol/L, pH is respectively 4.0,4.5,5.0,5.5 acetic acid-acetic acid
In sodium cushioning liquid, respectively circulated 5 minutes with 5 mul/min of flow velocity.The pH of the maximum solution of signal intensity is at the end of circulation
5.5, so also using this pH cushioning liquid during rear protein covalent bond.
4th, circulated 30 minutes with EDC and NHS mixed solution, both concentration are respectively 0.01 and 0.002 grams per milliliter, stream
Speed is 5 mul/min.
5th, using 0.01 grams per milliliter, the HSA solution that pH is 5.0, with 5 mul/min of flow rates 20 minutes.
6th, with HBS-EP+ buffer solutions (containing 10mmol/LHEPES, 0.15mol/L sodium chloride, 3mmol/L EDTA,
PH is for the solution of 8% (volume fraction) pyridine of addition in 7.4) during P20,25 DEG C of the surfactant of 0.005% volume fraction
Diluent, pure pyridine is dissolved in 10mmol/L by rutin, be then diluted to 0.06 with this diluent, 0.10,0.15,0.20,
0.25th, 0.30mmol/L, dilutes the pure pyridine solution for being free of rutin during dilution with same ratio every time.
7th, the measure of protein and rutin interaction process.
Circulation solution in SPR instrument pipelines is replaced with to rutin diluent used in the 6th step.It will be obtained in 6th step
Each concentration rutin dilution after solution with it is corresponding respectively without rutin dilution pyridine solution according to by it is dilute to dense, elder generation
The order of rutin is sequentially sent to SPR instruments and analyzed after pyridine.Sample flow rate is 5 mul/min, is maintained 5 minutes after sample introduction,
Hereafter with the SDS solution flushing lines of mass fraction 5%, then solution in pipeline replaces with the 6th step to retarder thinner used,
Analyzed next time.HSA with containing rutin each sample solution interaction gained spr signal as shown in Fig. 2 HSA with
Spr signal obtained by the interaction of each pyridine dilute solution without rutin is as shown in Figure 3.In the present embodiment, c1=
0.06mmol/L, c2=0.10mmol/L, c3=0.20mmol/L, c4=0.25mmol/L, c5=0.30mmol/L；R1=
129RU, R2=178RU, R3=225RU, R4=225RU, R5=224RU；r1=118RU, r2=161RU, r3=199RU, r4=
198RU, r5=193RU.
8th, data analysis.
Protein used is tried to achieve according to following steps, and the equilibrium constant of association reaction occurs with rutin.
A, record each ciWhen RiAnd ri；Described in the 7th step；
B, with each 1/ (Ri-ri) to 1/ciMapping, 1/ciFor abscissa, 1/ (Ri-ri) it is ordinate, as shown in Figure 4；
C, in Fig. 4 each data point carry out linear fit, gained equation be y=3.84 × 10-3x+2.02×10-2, line
Property coefficient is 0.99.With intercept divided by slope, it is 5.3mmol to obtain rutin with the binding constant that HSA interacts-1L, i.e.,
5.3×103mol-1·L。
Claims (10)
1. a kind of determine rutin and the surface plasma body resonant vibration analysis method of protein binding level, comprise the following steps：
1) surface carboxyl groups modification is carried out to clean sensing chip；
2) pH value of the buffer solution for solubilized target albumen is screened；
3) carboxyl that censorchip surface is modified is activated；
4) target protein is dissolved in step 2) in selected pH buffer solution, pass through step 3) chip surface of activation, will
Target protein is covalently bound on chip；
5) it is c to prepare rutin concentration1、c2、…、ciSerial solution, wherein i be natural number；
6) by concentration by dilute to dense order by step 5) in the rutin solution feeding surface plasma body resonant vibration instrument prepared, pass through
Step 4) sensing chip after processing, each concentration is recorded for ciRutin solution sample introduction and signal stabilization after signal value Ri,
Meanwhile, record does not contain the signal value r after rutin but other compositions identical contrast solution sample introduction and signal stabilizationi；
7) data analysis：Calculate each 1/ciWith 1/ (Ri-ri), and with 1/ (Ri-ri) to 1/ciMapping, 1/ciFor abscissa, 1/
(Ri-ri) it is ordinate；Linear fit is carried out to data point in figure, the intercept of gained equation is designated as b, and slope is designated as a, then combined
The equilibrium constant of reaction is b/a, the inverse of unit concentration unit used in experiment.
2. analysis method as claimed in claim 1, it is characterised in that in step 1) in, the sensing chip is gold-plated glass
Piece, using by concentrated ammonia liquor, 30% hydrogen peroxide, water by volume 1:1:5 solution mixed are cleaned, then with washing
It is net and dry.
3. analysis method as claimed in claim 1, it is characterised in that in step 1) surface carboxyl groups modification is carried out to chip
Method is to install chip into surface plasma body resonant vibration instrument, then flows through chip list using the solution for meeting following condition
Face：Solute is the straight-chain carboxylic acid that a kind of sulfydryl replaces, and wherein sulfydryl is located at another end of carbochain away from carboxyl, carbon chain lengths
For containing three to 20 carbon atoms, solute concentration is 2mmol/L to 50mmol/L, solvent is water and/or ethanol；In the process of circulation
Monitoring spr signal in real time, the currency after sample introduction untill spr signal no longer changes, but at least 1 minute, solution stream
Speed is 0.1 mul/min to 1 ml/min.
4. analysis method as claimed in claim 1, it is characterised in that in step 2) relatively more different pH protein solution flows through carboxylic
The change of spr signal during the chip of base modification：If in the several pH value attempted, under the used currency, each solution
Spr signal is set to reach stabilization, then signal differs the pH of maximum solution with baseline after selection is stable；If in the circulation used
Between under each solution signal is reached stabilization, then the pH of selection signal change most fast solution；If part reaches steady component
Not up to stable, then circulation is selected to complete the pH that time-ofday signals differ maximum solution with baseline.
5. analysis method as claimed in claim 1, it is characterised in that step 3) following method I or II are used to chip surface
The carboxyl of modification is activated：
Method I：The solution for meeting following condition is set to flow through chip surface：Solute is N- (3- dimethylamino-propyls)-N '-second
Base carbodiimide hydrochloride and N- hydroxysuccinimides, both concentration are 0.0001 to 0.5 grams per milliliter, and the former concentration
It is not less than the latter's concentration, flow velocity is between 0.1 mul/min between 1 ml/min, and the currency is no longer changed with spr signal
It is defined, but at least 1 minute, chip area temperature is a value between 0 DEG C to 60 DEG C；
Method II：Successively make N- (3- dimethylamino-propyls)-N '-ethyl-carbodiimide hydrochloride solution and N- maloyls sub-
Amine aqueous solution flows through chip surface respectively, and respective concentration is 0.0001 to 0.5 grams per milliliter, and flow velocity is between 0.1 mul/min
It is defined, but is each at least 1 minute, core between 1 ml/min, the currency is no longer changed by respective spr signal
Panel region temperature is a value between 0 DEG C to 60 DEG C.
6. analysis method as claimed in claim 1, it is characterised in that step 4) by the target protein second that pH is 4 to 5.5
Acid-sodium acetate buffer dissolving, Na ion concentration is not higher than 50mmol/L, and target protein concentration is 1 nanograms/milliliter to 50 millis
Between grams per milliliter, be allowed to flow through chip surface, chip area temperature is a value between 0 DEG C to 60 DEG C, flow velocity between
0.1 mul/min is defined between 1 ml/min, the currency is no longer changed by spr signal, but at least 1 minute.
7. analysis method as claimed in claim 1, it is characterised in that step 5) first rutin is dissolved in pure pyridine, obtain
Concentration is the pyridine solution of 1~50mmol/L rutin, and then the PBS bufferings of the pyridine with the addition of 3%~15%v/v are molten
One of liquid, Tris cushioning liquid or HEPES cushioning liquid are as retarder thinner, and pH is 6.6~8.5 at 25 DEG C, is diluted with it
The pyridine solution of rutin, obtains rutin concentration for c1、c2、…、ciSerial solution；Meanwhile, with the retarder thinner with same ratio
Pure pyridine is diluted, the pyridine solution of various concentrations is obtained as contrast solution.
8. analysis method as claimed in claim 7, it is characterised in that be no more than 5wt% containing concentration in the retarder thinner
Additive.
9. analysis method as claimed in claim 7, it is characterised in that the retarder thinner is to contain 10mmol/L
HEPES, 0.15mol/L sodium chloride, 3mmol/L EDTA, 0.005%v/v surfactant P20, pH is 7.4 at 25 DEG C
8%v/v pyridines are with the addition of in buffer solution.
10. analysis method as claimed in claim 7, it is characterised in that step 6) by step 5) solution prepared is according to first right
Order according to rutin solution after solution is sequentially sent to surface plasma body resonant vibration instrument and analyzed, and flow velocity is between 0.1 mul/min
To between 1 ml/min, between 0 DEG C to 60 DEG C of chip area temperature control, equal tracer signal after each sample introduction until no longer change,
And at least maintain 1 minute.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603828A (en) * | 2004-10-20 | 2005-04-06 | 国家海洋环境监测中心 | Surface plasma resonance rapid detection method for paralytic shellfish poisoning |
| CN104962633A (en) * | 2015-07-03 | 2015-10-07 | 南京理工大学 | Nucleic acid detection method based on surface plasmon resonance technology |
| US20160011216A1 (en) * | 2014-07-10 | 2016-01-14 | International Business Machines Corporation | Biosensors including surface resonance spectroscopy and semiconductor devices |
| US20170176450A1 (en) * | 2012-02-27 | 2017-06-22 | The University Of Kansas | Systems and methods for identifying protein stabilizers |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603828A (en) * | 2004-10-20 | 2005-04-06 | 国家海洋环境监测中心 | Surface plasma resonance rapid detection method for paralytic shellfish poisoning |
| US20170176450A1 (en) * | 2012-02-27 | 2017-06-22 | The University Of Kansas | Systems and methods for identifying protein stabilizers |
| US20160011216A1 (en) * | 2014-07-10 | 2016-01-14 | International Business Machines Corporation | Biosensors including surface resonance spectroscopy and semiconductor devices |
| CN104962633A (en) * | 2015-07-03 | 2015-10-07 | 南京理工大学 | Nucleic acid detection method based on surface plasmon resonance technology |
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