CN101271066A - Method for researching interaction of medicament and biomolecule by surface plasma body resonance sensor - Google Patents
Method for researching interaction of medicament and biomolecule by surface plasma body resonance sensor Download PDFInfo
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Abstract
The invention belongs to the field of a method of researching the interaction between medicine and biomolecules with a wavelength detecting surface plasmon resonance sensor. A wavelength detecting SPR analyzer is adopted; a glass prism is selected as a sensing element of the sensor; a vacuum film coating method is used to coat a layer of about 50nm thick gold or silver film on a sensing face of the prism; mercaptopropionic acid (MPA) solution, 1-ethyl-3-(3-dimethylamino propyl) carbodiimide hydrochloride (EDC for short), N-hydroxysuccinimide (NHS for short) solution and serum albumin solution are subsequently injected into a flow-through cell at the bottom of a sensing film; when the resonant wavelength is basically stable, PBS buffer solution is used to repeatedly flush the surface of the sensor until the resonant wavelength does not change any more; the medicine is respectively diluted into n portions of solution in different concentration by the PBS buffer solution which are subsequently injected into the flow-through cell, and the changes of the resonant wavelength as the time are recorded. Through calculation, the kinetic constant, the thermodynamic constant and the combining percent after interaction of the medicine and the protein molecules.
Description
Technical field
The invention belongs to wavelength detection type surface plasmon resonance (Surface plasmon resonance, SPR) the method field of sensor research medicine and bio-molecular interaction, particularly the wavelength detection type SPR analyser and the application process thereof of the development of this seminar.
Background technology
Since Liedberg etc. was used for the SPR technology chemistry and biology sensor research field, spr sensor became the research focus of international sensor field gradually.Carry out the technology emerge in multitude of functional study in recent years on molecular level, wherein the SPR technology is more noticeable.Especially since the nineties Pharmacia company in last century (now being BIAcore AB company) was with this technology commercialization, the research that the SPR technology is used for bio-molecular interaction had obtained swift and violent development.In a few years, the document that uses this new technology to deliver has almost comprised the every field of modern biomedical, makes the research of interaction of molecules obtain unprecedented breakthrough, and then has promoted the progress of the every field of life science comprehensively.Because having in real time monitoring reaction dynamic process, sample, the SPR technology need not characteristics such as mark, sensitivity are higher, no background interference, be mainly used in the interaction between the big molecule of research, can obtain the bonding information in each step between the reactant molecule, measure kinetic constant, in bio-science field is used, make significant progress.According to rough statistics, in the article that field of biosensors is delivered, nearly 70% has reported about spr sensor and has been applied to detection to reacting between biomolecule.
All kinds of SPR commercial apparatuss of having reported all are to adopt fixed wave length, measure the mode of operation that the resonance angle changes, this mode of operation perhaps needs a mechanical driving device and changes incident angle of light, perhaps need disperse function, measure the variation of angle by pointolite.The former has a movable member in instrument, the angular range that the latter measures is less, has limited the application of method.The commercially available prism-type spr sensor instrument of this class costs an arm and a leg, and should not popularize.The wavelength detection type SPR analyser of design and assembly realizes that easily multi-wavelength detects simultaneously voluntarily, and the wavelength coverage that detects is unrestricted, so not only make the scope of sensor surface detected material quality unrestricted, the scope that is variations in refractive index is unrestricted, and is a significant benefit to research intermolecular or molecule and tissue (as cell) interphase interaction; Secondly, owing to be used for the device comparative maturity that wavelength is selected now, and the device that has has very high wavelength resolution ability, so be expected to further improve the sensitivity of SPR analyser.This instrument is particularly suitable for the research of intermolecular interaction, and is significant for effect, drug screening and the pharmaceutical chemical development of research medicine and biomolecule, and also significant to the development and the application of spr sensor method, apparatus.
The traditional medicine and the research method of protein bound have: equilibrium dialysis and ultrafiltration.Wherein equilibrium dialysis is classical reference method, owing to need set up binding equilibrium, uses this method length consuming time (several hrs or several days).The ultrafiltration rule is brought very big inconvenience owing to often carrying out radioactivity or fluorescence labeling.All there is the absorption of film to medicine in these two kinds of methods, and Donnan effect problems such as (because albumen and medicine are all electrically charged, making that the free drug concentration of film both sides is unequal), are difficult to accurate detection for the free drug concentration of high-bonding-ratio medicine.Additive method also has immunoassay, liquid phase chromatography, fluorescent quenching method and capillary electrophoresis.Immunoassay length consuming time wherein, and can only be used for qualitative, sxemiquantitative, can't satisfy the requirement of profound medicine and protein bound research; Liquid phase chromatography needs not only that sample size is big, post absorption is serious, and often needs to add the improvement solvent, can not accurately estimate medicine and protein bound effect under the physiological condition; When the site of medicine and albumin bound during, use the fluorescent quenching method this moment and just can not detect combining of medicine and albumen away from tryptophane; The topmost shortcoming of capillary electrophoresis is that its sensitivity is not high.
In recent years, SPR has been widely used in the combination research of medicine-albumen.A series of drug effects on fixing albumen, are determined the situation that combines of medicine and albumen according to the medicine and the spr signal intensity of albumen effect.In the application of SPR, general molecule one end that requires to do sensitive membrane for can with the reactive group of gold or silver-colored bonding, and its other end is the reactive group with molecular recognition function.The sulfydryl end of mercaptopropionic acid can connect with golden symphysis, and another terminal carboxyl can be used as reactive group, connects antibody molecule with it, and the result is good.
Summary of the invention
The objective of the invention is to adopt wavelength detection type spr sensor, the interaction of research drug molecule and biomolecule.
The present invention comes to be realized by the following technical programs: adopt wavelength detection type spr sensor, the method for research drug molecule and bio-molecular interaction relates to the following step:
1. adopt the wavelength detection type SPR analyser of this seminar development: form by light source 1, light-conducting system 2,3,4,10,11, sensing element 5,6, flow cell 7,8,9, spectral detection system 12 and data handling system;
2. the following step is adopted in the preparation of sensor: in wavelength detection type SPR analyser, select the sensing element of glass prism 5 as sensor for use; Adopt Vacuum Coating method, at thick gold or the silverskin of the sensitive face about 50nm of plating of prism 5, as the sensing membrane 6 of sensor;
3. in the flow cell 7 of sensing membrane 6 bottoms, inject mercaptopropionic acid (MPA) solution, treat that MPA after golden film surface self assembly is finished, is injected into flushing repeatedly in the flow cell 7 with phosphate (PBS) buffer solution, to remove the non-specific binding of sensor surface;
4. after treating that resonant wavelength no longer changes, in flow cell 7, inject 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide hydrochloride (1-Ethyl-3-(3-dimethylaminopropyl)-carbodimide hydrochloride, C
8H
17N
3HCl, molecular weight are 191.7, are called for short EDC) and N-hydroxy-succinamide (N-Hydroxysuccinimide, C
4H
5NO
3, molecular weight is 115.1, is called for short NHS) and solution, observe SPR spectrum over time, when resonant wavelength is basicly stable, washes sensor surface to resonant wavelength repeatedly with PBS buffer solution and no longer change;
5. injection serum albumin solution, the record resonant wavelength is situation over time, after seralbumin formed stable unimolecular layer, when resonant wavelength was stablized, be injected into PBS buffer solution to wash repeatedly in the flow cell 7 to resonant wavelength and no longer change this moment;
6. be n variable concentrations with the dilution of PBS buffer solution respectively with medicine, be injected into flow cell 7 successively, the record resonant wavelength over time.After each sample monitoring finishes, all inject PBS buffer solution flushing flow cell 7.
7. pass through the response signal Δ λ of drug concentrations [D], wavelength variations Δ λ, wavelength variations
Max,, can calculate the association rate constant k that reflects drug molecule and bio-molecular interaction by following equation (1)
aWith the rate constants k of dissociating
d
dΔλ/dt＝k
a[D](Δλ
max-Δλ)-k
dΔλ (1)
The present invention can also introduce in order to improve the golden nanometer particle of transducer sensitivity after the MPA self assembly finishes.
The present invention can also introduce in order to the albumen outside the pure albumen of one or more layers dehematize that improves transducer sensitivity after the MPA self assembly finishes, and constitutes sandwich sandwich sensor.
Instrument of the present invention is made up of light source, light-conducting system (lens, polaroid, optical fiber), sensing element (glass prism, sensing membrane), flow cell, spectral detection system (CCD detecting device) and data handling system: with halogen tungsten lamp or light emitting diode as light source, the white light that sends from light source becomes parallel polarized light after through parallel polarization light pipe (being made up of two lens and a polaroid), to the side of prism, light arrives the bottom of prism after reflecting with certain angular illumination.Adopt Vacuum Coating method, at thick gold or the silverskin of the sensitive face about 50nm of plating of prism, adopt the molecule self-assembly method to be coated with one deck has the specific bond ability to determinand molecule in metallic film surface, form sensitive membrane, light is in the experiences total internal reflection at the interface of prism and metal, another side from prism reflects away then, be coupled into light transmitting fiber by next lens focus, by optical fiber flashlight is transferred to grating and charge-coupled device (CCD) detecting device again, when containing the change of determinand and testing concentration in the solution, resonant wavelength has significant change, in view of the above determinand is studied.Experimental technique is that sensor is fixed on above the flow cell mouth, the flow injection sample introduction, sensor plated film one side contacts with analyzed test solution when sample passes through flow cell, polychromatic light is radiated on the prism after by polaroid and lens, resonate, produce strong resonance absorption,, measured matter is studied by drawing intensity of reflected light with the wavelength change curve.
The drug molecule of the present invention's research and the association rate constant k of bio-molecular interaction
aWith the rate constants k of dissociating
dComputing method as follows: Δ λ is the variation of resonant wavelength in the aforementioned equation (1), i.e. the signal of SPR, it has represented the concentration of sensor surface seralbumin (P) and medicine (D).D Δ λ/dt is the rate of change of spr signal.Δ λ max is the response signal of sensor surface bound drug when saturated, promptly is equivalent to the Cmax of the PD compound that sensor surface forms.The integrated form of equation (1) is: Δ λ={ k
a[D] Δ λ
Max/ (k
a[D]+k
d) { 1-exp (([D]
k
a+k
d)t) (2)
Dissociation process can be used following The Representation Equation:
dΔλ/dt＝-k
dΔλ (3)
Its integrated form is:
Δλ＝Δλoe
-kd(t-to)
lnΔλ/Δλo＝-k
d(t-t
o) (4)
T in the formula
oBe the time of reaction beginning, its pairing spr signal is Δ λ
o, and Δ λ is the spr signal of any moment t.According to experimental data and equation (1)-(4), can obtain dynamics data k
a, k
dWith thermodynamic data binding constant K
AWith the constant K of dissociating
D
Description of drawings
Fig. 1 is the structural representation of wavelength detection type SPR analyser; 1 is light source, and 2,3 and 10 is lens, and 4 is polaroid, and 5 is glass prism, and 6 is sensing membrane, and 7 is flow cell, and 8 is injection port, and 9 are the waste discharge mouth, and 11 is optical fiber, and 12 is the CCD detecting device.
Fig. 2 is the flow cell structural representation; 5 is glass prism, and 6 is sensing membrane, and 7 is flow cell, and 8 is injection port, and 9 are the waste discharge mouth, and 13 are water-bath, and 14 are the water-bath water inlet, and 15 are the water-bath water delivering orifice.
Fig. 3 is the sensor connection diagram, also is Figure of abstract; 6 is sensing membrane, and 16 is substrate of glass (prism bottom surface), and 17 is mercaptopropionic acid and NHS, EDC decorative layer, and 18 is the seralbumin layer, and 19 is the drug molecule articulamentum.
Fig. 4 is not for containing MPA and containing the SPR spectrum that obtains when MPA solution is introduced sample cell; 20 is the SPR spectrum that obtains when not containing MPA; 21 is the SPR spectrum that obtains when containing MPA.
Embodiment
Embodiment 1: antibiotics and human serum albumins Study of Interaction
Microbiotic claims antibiotic again.Synthetic by some microorganisms, as to suppress or kill some pathogen chemical substance.Microbiotic is a kind of secondary metabolite of microorganism, can play inhibition or killing action to another kind of microbial growth and metabolism when a small amount of low concentration uses.Have now found that more than 400 kind of microbiotic, various antibiotic antibiotic mechanism and effect have nothing in common with each other.Hinder the synthetic of bacteria cell wall as penicillin and bacillus Toplink; The polypeptide antibiotics can destroy the plasma membrane of bacterium; But the synthetic or interfere RNA of Profilin such as chloromycetin, streptomysin matter is synthetic etc.The antibiotic reach that has is little, and is only effective to gram-positive bacterium as penicillin, is called narrow spectrum antibiotic.Some antibiotic coverage is wider, as tetracycline most bacteriums is all had inhibiting effect, is called broad-spectrum antibiotics.The about kind more than 100 of microbiotic of widespread use at present all generally is used for medical treatment as penicillin, streptomysin, gentamicin etc.Kasugarnycin, jinggangmycin have been widely used in the control of paddy rice germ on agricultural, and blasticidin-S is used to prevent and treat rice blast, and gibberellin is used to growth that promotes plant or the like.But will be when using microbiotic at illness, can not abuse.The microbiotic that has can produce some spinoffs, all can cause deafness as continuous use streptomysin, gentamicin, and the chloromycetin of overdose can cause leukopenia etc.Also some people have allergic reaction to certain antibiosis, when serious even fatal risk arranged.The present invention uses the wavelength detection type SPR analyser that designs voluntarily and assemble, adopt the method for amino coupling connection that seralbumin is fixed on sensor surface, studied ospen K, Oxacillin, the Amoxicillin, cefoperazone, Cefotaxime Sodium, Enoxacin, the interaction of seven kinds of antibiotic medicines of Norfloxacin and human serum albumins (HSA).Measure the kinetic constant and the equilibrium constant of reaction, calculated their bonding percent simultaneously.
1. instrument: the wavelength detection type SPR analyser that adopts the development of this seminar;
Reagent:
3-mercaptopropionic acid (MPA), NHS, EDC
Ospen K, Oxacillin, Amoxicillin, cefoperazone, Cefotaxime Sodium, Enoxacin, Norfloxacin standard items
Human serum albumins, bovine serum albumin(BSA)
Other all reagent are homemade analytical reagent
2. the following step is adopted in the preparation of sensor: in wavelength detection type SPR analyser, select the sensing element of the bottom surface of right-angle glass prism as sensor for use; Adopt Vacuum Coating method, at thick gold or the silverskin of the sensitive face about 50nm of plating of prism, as the sensing membrane of sensor;
3. will inject PBS buffer solution in the sample cell, the record resonant wavelength, with the MPA solution injection sample cell of 10mmol/L, by the displacement of observation SPR resonant wavelength, thus monitoring MPA and the golden dynamic process that combines.Fig. 4 is not for containing MPA and containing the SPR spectrum that obtains when MPA solution is introduced sample cell.After will containing the solution injection sample cell of MPA, As time goes on, resonant wavelength constantly moves to the long wave direction, and assembling is comparatively rapid in a few minutes of beginning.In 5min, SPR spectral resonance wavelength displacement value reaches 99% of total amount, about 30min the assembling complete substantially, the maximum displacement value of resonant wavelength is 11.67nm.Substantially no longer change with PBS buffer solution flushing sensor surface to resonant wavelength, illustrate that the sulfydryl end of MPA is very stable with the S-Au key that gold forms.
4. in flow cell, add EDC and the NHS mixed solution that concentration is 40mg/mL, but the reaction of EDC catalysis MPA and NHS makes the c-terminus acidylate of MPA, is beneficial to sero-abluminous connection.
5. after the assembling fully of MPA modified membrane, wash sensor surface repeatedly, to remove specific adsorption with the PBS damping fluid.Then seralbumin is injected sample cell.Because of after the c-terminus acidylate of MPA to seralbumin Fc section can specificity combination, can make seralbumin directed self assembly on sensitive membrane.The effects human serum albumins (HSA) concentration it is carried out the influence of directed self assembly on MPA.HSA is mixed with the solution that concentration is 0.05mg/mL, 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, and the result (12h) of the HSA of variable concentrations self assembly on MPA sees Table 1.Take all factors into consideration, selecting the concentration of HSA is 0.1mg/mL.
The result (12h) of the HSA of table 1 variable concentrations self assembly on MPA
When seralbumin after being self-assembled into stable molecular film on the sensitive membrane, wash flow cell repeatedly with the PBS damping fluid, when treating that resonant wavelength no longer changes, in flow cell, inject the ospen K of variable concentrations successively, Oxacillin, the Amoxicillin, cefoperazone, Cefotaxime Sodium, Enoxacin or Norfloxacin (all preparing) with PBS, the resonant wavelength shift value of monitoring in real time and record SPR spectrum, minute is 30min.
7. according to monitoring gained data, also calculate by analysis, can obtain kinetic constant, thermodynamic equilibrium constant and the bonding percent (seeing Table 2) of bonding between these medicines and HSA and the BSA.
The thermodynamic equilibrium constant of bonding and bonding percent between table 2 medicine and the HSA
These 7 kinds of antibiotics and HSA have combining in various degree, the bonding percent scope of medicine and HSA is 91.241.1%, the order of their binding ability is ospen K＞cefoperazone＞Cefotaxime Sodium＞Oxacillin＞Amoxicillin＞Enoxacin＞Norfloxacin.
Embodiment 2: anthraquinone class medicine and human serum albumins Study of Interaction
Rheum officinale is one of four big Chinese medicines of China, and the beginning is stated from Shennong's Herbal.The output of China's rheum officinale accounts for the No. 1 in the world, and optimal quality.China has begun to use rheum officinale before 4600.At present existing nineteen state-promulgated pharmacopoeia has been recorded rheum officinale and has been used as official drug.Rheum officinale mainly contains anthraquinone derivatives, and its content is 3～5%, and a part is a free state, as Rhein, Chrysophanol, archen, aloe-emodin etc.; Major part is a bonding state, and what wherein belong to the anthraquinone glucoside has Rhein, glycoside, a chrysophanol monoglucoside etc.Have purgating heat and bowels, removing pattogenic heat from the blood and toxic material from the body, pursue the function that the stasis of blood is stimulated the menstrual flow.Be used for real hot constipation, stagnant stomachache, to rush down dysentery not well, jaundice with damp-heat pathogen, blood-head tell that nosebleed, hot eyes, pharynx are swollen, abdominalgia with intestinal abscess, the swollen furunculosis of carbuncle, hemostasis through close, illness such as traumatic injury.Modern clinical can be used for treating epidemic meningitis, lobar pneumonia, acute biliary infection, acute parotidits, acute appendicitis, Fiedeler's disease type hepatitis, chordapsus, bacillary dysentery, hemorrhage of digestive tract, sphagitis, parulis, dermatitis, eczema, gonorrhoea, herpes zoster etc.Research anthraquinone class medicine and sero-abluminous interactional have fluorescent spectrometry, ultraviolet spectroscopy.The present invention uses the wavelength detection type SPR analyser that designs voluntarily and assemble, with 3-mercaptopropionic acid (MPA) as the fixing articulamentum of HSA, study the principal ingredient archen in the rheum officinale, Chrysophanol, reached the interaction of Physcion and HSA, and calculated their kinetic constant, thermodynamic equilibrium constant and bonding percent respectively.
Method is with embodiment 1.The results are shown in Table 3.
The kinetic constant that combines of table 3 anthraquinone class medicine and HSA, thermodynamic equilibrium constant and in conjunction with percent
Embodiment 3: panaxoside and seralbumin Study of Interaction
Genseng is commonly called as " wooden club ", belongs to the Araliaceae per nnial herb, is one of important special product of China, also is the valuable ingredient of traditional Chinese medicine that has won fame both at home and abroad, and is called " kings of hundred grass " by people, belongs to one of northeast " Triratna ".Ginseng, stem, leaf, flower and fruit are rich in multiple panaxoside, and ginseng contains volatile oil 0.12% in addition, and cauline leaf contains volatile oil 0.13%, and flower contains volatile oil 0.29%.The nonpolar part of ginseng contains Panaxynol, α-panacen, β-panacen and sterol compound.Genseng also contains several amino acids, vitamin, multiple organic acid, flavone compound and sugar such as lysine, histidine, arginine.Through traditional Chinese medical science clinical trial for many years, genseng can be enriched blood and be fostered the spirit of nobility, Gu Jin gives birth to liquid, it is neural to regulate, make eye bright happily, intelligence promoting and tranquilization, blood sugar lowering, be good for the stomach, diuresis etc.; For the valetualinary patient of treatment, very effective.Cure mainly nervous breakdown, various neuropathy, the sick imbalance of autonomic nerve, neurasthenia sexualis, hypophrenia, anaemia, diabetes, stomach trouble, hepatopathy, and the disease of cardiovascular system etc.In addition, an amount of clothes for a long time can also make the people increase resistibility to various virulence factors, and human body is not produced any spinoff and infringement.Pharmacological action in most of year of genseng is decided by panaxoside.Be separated to so far ginsenoside monomer have 40 surplus kind, it has very big influence to some institutional frameworks of human body: can the neuroprotective cell avoid the infringement of some hazardous compounds; Can usually regulate neural transmission by reducing nerve conduction; Play a part aspect the drug-induced sleep very big; Also can cancer cell be exerted an influence the growth of anticancer, or metastasis cancer cell element by different mechanism.The present invention uses the wavelength detection type SPR analyser that designs voluntarily and assemble, with 3-mercaptopropionic acid (MPA) as fixing sero-abluminous articulamentum, 6 kinds of panaxoside monomer (Rb1 have been studied, Rb2, Rb3, Rc, Rd, Re) respectively with the interaction of human serum albumins (HSA) and bovine serum albumin(BSA) (BSA).Calculate their kinetic constant, thermodynamic equilibrium constant and bonding percent respectively, and compared different panaxoside monomers and sero-abluminous binding ability.
Method is with embodiment 1.The results are shown in Table 4.
The kinetic constant that table 4 panaxoside combines with HSA and BSA, thermodynamic equilibrium constant and in conjunction with percent
Claims (3)
1. with the method for surface plasmon resonance sensor research medicine and bio-molecular interaction, it is characterized in that adopting the following step:
(1) the wavelength detection type SPR analyser that adopts this seminar to develop: form by light source (1), light-conducting system (2,3,4,10,11), sensing element (5,6), flow cell (7,8,9), spectral detection system (12) and data handling system;
(2) the following step is adopted in the preparation of sensor: in wavelength detection type SPR analyser, select the sensing element of glass prism (5) as sensor for use; Adopt Vacuum Coating method, at thick gold or the silverskin of the sensitive face about 50nm of plating of prism (5), as the sensing membrane (6) of sensor;
(3) in the flow cell (7) of sensing membrane (6) bottom, inject mercaptopropionic acid (MPA) solution, treat that MPA is after golden film surface self assembly is finished, phosphate (PBS) buffer solution is injected into flushing repeatedly in the flow cell (7), to remove the non-specific binding of sensor surface;
(4) treat that resonant wavelength no longer changes after, in flow cell (7), inject 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide hydrochloride (1-Ethyl-3-(3-dimethylaminopropyl)-carbodimide hydrochloride, C
8H
17N
3HCl, molecular weight are 191.7, are called for short EDC) and N-hydroxy-succinamide (N-Hydroxysuccinimide, C
4H
5NO
3, molecular weight is 115.1, is called for short NHS) and solution, observe SPR spectrum over time, when resonant wavelength is basicly stable, washes sensor surface to resonant wavelength repeatedly with PBS buffer solution and no longer change;
(5) inject serum albumin solution, the record resonant wavelength is situation over time, after seralbumin formed stable unimolecular layer, when resonant wavelength was stablized, be injected into PBS buffer solution to wash repeatedly in the flow cell (7) to resonant wavelength and no longer change this moment;
(6) be n variable concentrations with the dilution of PBS buffer solution respectively with medicine, be injected into flow cell (7) successively, the record resonant wavelength after each sample monitoring finishes, all injects PBS buffer solution flushing flow cell (7) over time.
(7) the response signal Δ λ by drug concentrations [D], wavelength variations Δ λ, wavelength variations
Max,, can calculate the association rate constant k that reflects drug molecule and bio-molecular interaction by following equation (1)
aWith the rate constants k of dissociating
d
dΔλ/dt＝k
a[D](Δλ
max-Δλ)-k
dΔλ (1)。
2. according to the method for claim 1, it is characterized in that the MPA self assembly finishes after, introduce in order to improve the golden nanometer particle of transducer sensitivity.
3. according to the method for claim 1, it is characterized in that the MPA self assembly finishes after, introduce in order to the albumen outside the pure albumen of one or more layers dehematize that improves transducer sensitivity, constitute sandwich sandwich sensor.
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| CN105203504A (en) * | 2015-09-21 | 2015-12-30 | 清华大学深圳研究生院 | Method for improving sensitivity of surface plasmon resonance sensor |
| CN105203504B (en) * | 2015-09-21 | 2017-10-27 | 清华大学深圳研究生院 | A kind of method for improving surface plasma resonance sensor sensitivity |
| CN108139321A (en) * | 2015-10-12 | 2018-06-08 | 通用电气健康护理生物科学股份公司 | For determining the method for the instrument of surface plasma resonance biosensor system dependence parameter |
| US11567005B2 (en) | 2015-10-12 | 2023-01-31 | Cytiva Sweden Ab | Method in a surface plasmon resonance biosensor system |
| CN106918543A (en) * | 2017-02-28 | 2017-07-04 | 同济大学 | A kind of device and method for detecting single gold nano grain surface biomolecules |
| CN107356561A (en) * | 2017-06-29 | 2017-11-17 | 暨南大学 | Surface plasma resonance sensor of tungsten disulfide enhanced sensitivity and preparation method thereof |
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