CN108627646A - One kind being based on two dimension MoS2Nanometer sheet and carcinomebryonic antigen aptamers structure biological sensor and for detecting carcinomebryonic antigen - Google Patents

One kind being based on two dimension MoS2Nanometer sheet and carcinomebryonic antigen aptamers structure biological sensor and for detecting carcinomebryonic antigen Download PDF

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CN108627646A
CN108627646A CN201810633861.1A CN201810633861A CN108627646A CN 108627646 A CN108627646 A CN 108627646A CN 201810633861 A CN201810633861 A CN 201810633861A CN 108627646 A CN108627646 A CN 108627646A
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carcinomebryonic antigen
aptamers
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卢革宇
赵连静
高原
刘方猛
闫旭
孙鹏
梁喜双
刘凤敏
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Jilin University
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    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57473—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving carcinoembryonic antigen, i.e. CEA

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Abstract

One kind being based on two dimension MoS2The biological sensor and the application in detecting carcinomebryonic antigen of nanometer sheet and carcinomebryonic antigen aptamers structure, belong to biological sensor technical field.The sensor on two kinds of material foundations based on building, first, having high-efficiency fluorescence quenching ability and having the two-dimentional MoS of good biological identification function to single stranded DNA, single stranded DNA and albumen composition2Nanometer sheet；Second is that having the characteristics that the carcinomebryonic antigen aptamers that specific recognition antigen, at low cost, stability is good, is easy to modification.The features such as biosensor has structure simple, quickly detects, high sensitivity, and specificity is good, of low cost, favorable repeatability.For biosensor constructed by the present invention under optimum optimizing condition, the linear detection range to carcinomebryonic antigen is 100pg/mL~100ng/mL, Monitoring lower-cut 34pg/mL.The biological sensor has good application prospect in terms of the screening of kinds of tumors, diagnosis and prognostic evaluation.

Description

One kind being based on two dimension MoS2The biological of nanometer sheet and carcinomebryonic antigen aptamers structure Sensor and for detecting carcinomebryonic antigen
Technical field
The invention belongs to biological sensor technical fields, and in particular to one kind being based on two dimension MoS2Nanometer sheet and cancer embryo Antigen aptamers structure biological sensor and for detecting carcinomebryonic antigen.
Background technology
Because the tumor cure rate in late period is very low, so it is to defeat an important method of tumour that early diagnosis, which is early controlled, and it is high Sensitive Detection tumor markers can greatly improve the accuracy rate of early diagnosis of tumor.Simply, tumour is detected quickly and sensitively Marker carcinomebryonic antigen is all extremely important for screening, diagnosis and a variety of gastroenteric tumors of prognosis evaluation.It is clinically used at present The method of carcinomebryonic antigen detection mainly has enzyme-linked immunosorbent assay, radioimmunoassay and chemiluminescence immunoassay Method, however these immunization methods, there is adult height, detection is slow, requires operating personnel's technology high, and detection process is cumbersome equal to be lacked Point.The advantages that biosensor is at low cost because of, quickly analysis simple with structure, high sensitivity and be concerned.Therefore it develops Biosensor for tumor-marker analyte detection has very important potential applicability in clinical practice.Biosensor is much to receive It is established on the basis of rice material, common material has gold nanoparticle, carbon nanotube, graphene, two-dimentional transition metal vulcanization Object, (M.C.Canbaz, the C.S. Simsek, M.K.Sezginturk, Electrochemical such as two-dimentional transition metal oxide biosensor based on self-assembled monolayers modified with gold nanoparticles for detection of HER-3,Analytica Chimica Acta,2014, 814,31-38.H.P.Peng,Y.Hu, P.Liu,Y.N.Deng,W.Peng,W.Chen,A.L.Liu,Y.Z.Chen, X.H.Lin,Label-free electrochemical DNA biosensor for rapid detection of mutidrug resistance gene based on Au nanoparticles/toluidine blue–graphene oxide nanocomposites. Sensors and Actuators B:Chemical 2015,207,269-276.).With the appearance of graphene, two-dimensional nano Material receives more and more attention.Compared with other materials, it is latent that two-dimensional metallic sulfide shows good biological applications Energy：1) higher surface-to-volume ratio, can load a variety of a large amount of functional moleculars, 2) stronger fluorescent quenching ability, 3) Have very strong absorption at near-infrared, 4) good photo-thermal effect, 5) ultra-thin transverse dimensions make it have luminescent properties, have used In research enzymatic activity and cell viability.Electrochemical immunosensor currently based on two-dimensional metallic sulfide structure divides for detecting Hydrogen peroxide is analysed, glucose, dopamine, DNA is more, and seldom for the biological sensor of protein type tumor markers. Therefore, structure is a kind of simple, quickly, at low cost, the biological sensor tool for detecting tumor markers of high sensitivity It is significant, and use two-dimentional MoS2 nanometer sheets with efficient quenching ability and good biological identification function and with cost Low, stability is good, be easy to modification the features such as aptamers be realizes this function admirable biosensor one kind effectively on the way Diameter.
Invention content
The purpose of the present invention is to provide one kind being based on two dimension MoS2The fluorescence of nanometer sheet and carcinomebryonic antigen aptamers structure Biosensor and for detecting carcinomebryonic antigen.The sensor on two kinds of material foundations based on building, first, with efficient Fluorescent quenching ability and there are two-dimentional MoS2 nanometers of good biological identification function to single stranded DNA, single stranded DNA and albumen composition Piece；Second is that have specific recognition antigen, at low cost, stability is good, be easy to modification the features such as carcinomebryonic antigen aptamers.The life The features such as object sensor has structure simple, quickly detects, high sensitivity, and specificity is good, of low cost, favorable repeatability.It will Two-dimensional MoS2Nano material detects tumor markers for building biosensor, and simultaneously with aptamers replace antibody into The identification of row targeting proteins, and aptamers have specificity very well and selectivity to targeting proteins.Connect with aptamers in the present invention The fluorescent dye connect is texas Red (Texas Red), which under appropriate conditions can be with MoS2Nanometer sheet Fluorescence resonance energy transfer occurs.
It is of the present invention a kind of based on two dimension MoS2The biological sensing of nanometer sheet and carcinomebryonic antigen aptamers structure The preparation method of device, its step are as follows：
(1) 5 '-Texas Red (moral gram saxophone is red) of design carcinomebryonic antigen aptamers probe- ATACCAGCTTATTCAATT-3 ' send bio-engineering corporation to synthesize, and obtains aptamers probe powder；Its nucleotide sequence comes From in document：S.Su,M.Zou,H.Zhao,C.Yuan,Y.Xu,C.Zhang,L.Wang,C.Fan, L.Wang,Shape- controlled gold nanoparticles supported on MoS(2)nanosheets: synergistic effect of thionine and MoS(2)and their application for electrochemical label- free immunosensing,Nanoscale 2015,7,19129-35；
(2) by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Centrifuge tube is taken to be added (buffer solution contains the NaCl of a concentration of 100mM, a concentration of 5mM in 998 μ L, 10mM Tris-HCl (pH 7.4) buffer solution KCl and a concentration of 5mM MgCl2, solvent is deionized water), 2 μ L, 5 μM of aptamers probe solutions are added to the centrifugation Guan Zhong shakes mixing；Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, the cancer embryo for obtaining known concentration is anti- Original solution；The solution is added in centrifuge tube, final concentration of 0.1~100ng/mL of carcinomebryonic antigen；Then use 10mM, Tris-HCl (pH 7.4) buffer solution supplements volume to 1400~1900 μ L, and is incubated 1~3 hour in 37 DEG C of water-baths；
(3) by MoS2Nanometer sheet solution is added in the solution of step (2), and it is 2mL to make the volume of solution in centrifuge tube, It is reacted at room temperature after mixing 5~20 minutes；In centrifuge tube, MoS2Final concentration of 100~300 μ g/mL of nanometer sheet, aptamers The final concentration of 5nM of probe；
(4) solution that step (3) obtains is added in cuvette, carries out fluorescence spectrometry, it is strong then to draw fluorescence Relation curve between degree and carcinomebryonic antigen concentration；It is of the present invention based on two dimension MoS to obtain2Nanometer sheet and cancer embryo are anti- The biological sensor of former aptamers structure；Cuvette optical length 10mm, excitation wavelength 595nm, launch wavelength are 618nm；
(5) detection of unknown concentration carcinomebryonic antigen：Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, Obtain the carcinomebryonic antigen solution of unknown concentration；The carcinomebryonic antigen solution that known concentration in step (2) is replaced with the solution, is added to In centrifuge tube, the subsequent operation described in step (2) and step (3) is then carried out, fluorescence spectrometry is carried out, then according to step (4) concentration of carcinomebryonic antigen is calculated in the relation curve between the drafting fluorescence intensity in and carcinomebryonic antigen concentration.
The advantage of the invention is that：
1) by classical two-dimension nano materials MoS2Nanometer sheet is applied to structure biosensor system.Because two-dimensional MoS2Nanometer sheet not only has efficient fluorescent quenching ability, also has biometric identification capabilities, can identify aptamers, aptamers With the compound of albumen, so that the structure of sensor is simplified, eliminate the preparation process of multiple material, while keeping detection process fast Speed.
2) select aptamers that traditional protein antibodies is replaced to carry out the identification of targeting proteins so that sensor manufacturing cost It is lower, and stability is more preferable.
3) sensor has preferable selectivity and higher sensitivity, detection line 34pg/mL, linear detection range It is 0.1~100ng/mL.
4) response time is fast, and most of fluorescence is quenched in 1 minute, MoS in 5 minutes2Nanometer sheet is to aptamers probe Fluorescent quenching reaches stable state.
Operation principle：
Donors of the fluorophor Texas Red as fluorescence resonance energy transfer, excitation wavelength 595nm, transmitted wave A length of 618nm.MoS2Receptor of the nanometer sheet as fluorescence resonance energy transfer has efficient quenching ability.Fluorescent marker Aptamers pass through aptamers and MoS2Van der Waals force between nanometer sheet is adsorbed onto MoS2Nanometer sheet surface, to make fluorophor and MoS2Nanometer distance between commutator segments furthers so that fluorescence resonance energy transfer between the two occurs, and eventually leads to the glimmering of fluorophor Light is quenched.When, there are when carcinomebryonic antigen, carcinomebryonic antigen can be specifically bound with its aptamers, be caused in reaction system Aptamers conformational change, to make aptamers and MoS2Active force between nanometer sheet dies down, and makes it from MoS2Nanometer sheet surface is de- From making fluorophor and MoS in this way2Nanometer distance between commutator segments becomes larger, and fluorescence resonance energy transfer, final fluorogene cannot occur Fluorescence restore, the detection of carcinomebryonic antigen is realized by the variation of fluorescence signal.
Description of the drawings
Fig. 1 is based on MoS2The biological sensor principle for detecting carcinomebryonic antigen built on the basis of nanometer sheet is shown It is intended to.
Fig. 2 is that MoS is being added respectively in aptamers and aptamers/carcinomebryonic antigen2Fluorescent quenching curve after nanometer sheet；
Fig. 3 a are comparative example, the fluorescence spectra of 3 product of embodiment 1, embodiment 2 and embodiment；
Fig. 3 b are the fluorescence spectrum after comparative example, embodiment 1, embodiment 2 and 3 product of embodiment are incubated with carcinomebryonic antigen Figure.
Fig. 4 is the fluorescence intensity and addition cancer embryo of comparative example, embodiment 1, embodiment 2 and 3 product of embodiment at 608nm Fluorescence recovery strength (Δ F) comparison diagram after antigen at 608nm.
Fig. 5 a are fluorescence spectra of 2 product of embodiment after various concentration carcinomebryonic antigen is added；
Fig. 5 b are that the fluorescence for the biological sensor that the present invention is built restores ratio F/F0Between various concentration albumen Sensitivity analysis curve, illustration are that fluorescence restores ratio F/F0The calibration curve after logarithm is taken with various concentration albumen.F and F0Point It Wei not the front and back fluorescence intensity at 608nm of 2 product of embodiment addition carcinomebryonic antigen.
Fig. 6 is based on MoS2The selectivity of the biological sensor built on the basis of nanometer sheet and aptamers probe is illustrated Figure.BSA：Bovine serum albumin(BSA), IgG：Immunoglobulin G, ssDNA：Single stranded DNA.
As shown in Figure 1, aptamers by with MoS2Van der Waals force between nanometer sheet is adsorbed onto MoS2Nanometer sheet surface, to Make fluorophor and MoS2Nanometer distance between commutator segments furthers so that fluorescence resonance energy transfer between the two occurs, and eventually leads to The fluorescence of fluorophor is quenched.When, there are when carcinomebryonic antigen, carcinomebryonic antigen can occur special with its aptamers in reaction system The opposite sex combines, and leads to aptamers conformational change, to make aptamers and MoS2Active force between nanometer sheet dies down, and makes it from MoS2 Nanometer sheet surface is detached from, and fluorescence resonance energy transfer cannot occur, and the fluorescence of final fluorogene restores, and passes through fluorescence signal Change to realize the detection of carcinomebryonic antigen.
As shown in Fig. 2, MoS2Nanometer sheet is very fast to the quenching of aptamers probe and aptamers probe/carcinomebryonic antigen , reach balance in 5 minutes.
As shown in figure 3, in aptamers and MoS2Nanometer sheet system, aptamers/carcinomebryonic antigen and MoS2In nanometer sheet system, Different fluorescence spectrums is presented in comparative example and embodiment 1, embodiment 2 and embodiment 3, and compared with comparative example, in aptamers and MoS2Fluorescence in nanometer sheet system in embodiment 1, embodiment 2 and embodiment 3 is preferably quenched.When in aptamers/cancer embryo Antigen and MoS2In nanometer sheet system, comparative example and embodiment 1, embodiment 2 and 3 fluorescence of embodiment are by different degrees of extensive It is multiple..
As shown in figure 4, compared with comparative example, the fluorescence in embodiment 1, embodiment 2 and embodiment 3 is by more effectively sudden It goes out, and after the carcinomebryonic antigen that same concentrations are added, the fluorescence in comparative example, embodiment 1, embodiment 2 and embodiment 3 obtains Restore.Fluorescence wherein in embodiment 2 is in aptamers and MoS2Most fluorescence are quenched in nanometer sheet system, and suitable Ligand/carcinomebryonic antigen and MoS2Fluorescence is restored to the greatest extent in nanometer sheet system, shows best sensing characteristics.
As shown in Figure 5 a, in example 2 with the increase that carcinomebryonic antigen concentration is added, fluorescence signal is also gradually increasing By force, it is more than or equal to 80ug/mL in albumen concentration and obtains maximum fluorescence signal.
As shown in Figure 5 b, fluorescence restores ratio F/F in example 20Be added the increase of carcinomebryonic antigen concentration also by Cumulative big, as shown in being inserted into, when carcinomebryonic antigen concentration is in 0.1-100ng/mL ranges, fluorescence restores ratio F/F0It is dense with albumen The logarithm of degree is linearly related, linear equation F/F0=6.8408+6.78455log [CEA] (R2=0.9828).Show 2 sensor of embodiment has good sensitivity characteristic.
As shown in fig. 6, being separately added into control sample (BSA, IgG, ssDNA) and blank in same sensing system The carcinomebryonic antigen of group sample and various concentration finds that fluorescence intensity is all for the control sample (BSA, IgG, ssDNA) being added It is close with the fluorescence intensity of blank group sample (control), and after carcinomebryonic antigen is added, even the carcinomebryonic antigen of low concentration (1ng/mL), sensing system fluorescence intensity are apparently higher than each control group.Show that the biosensor has preferable selectivity.
Specific implementation mode
It elaborates below to the embodiment of the present invention, the present embodiment is carried out lower based on the technical solution of the present invention Implement, gives detailed embodiment and specific operating process, but protection scope of the present invention is not limited to following implementations Example.
Comparative example 1
1, aptamers probe is designed, the synthesis of Hai Sheng works bio-engineering corporation is served；Carcinomebryonic antigen aptamers probe sequence： 5 '-Texas Red (moral gram saxophone is red)-ATACCAGCTTATTCAATT-3 ', obtains aptamers probe powder；Wherein nucleosides Acid sequence comes from document：S.Su,M.Zou,H.Zhao,C.Yuan,Y.Xu,C.Zhang, L.Wang,C.Fan,L.Wang, Shape-controlled gold nanoparticles supported on MoS(2) nanosheets: synergistic effect of thionine and MoS(2)and their application for electrochemical label-free immunosensing,Nanoscale 2015,7,19129-35.
2, by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Centrifuge tube is taken to be added (buffer solution contains the NaCl of a concentration of 100mM, a concentration of 5mM in 998 μ L, 10mM Tris-HCl (pH 7.4) buffer solution KCl and a concentration of 5mM MgCl2, solvent is deionized water), 2 μ L, 5 μM of aptamers probe solutions are added to the centrifugation Guan Zhong shakes mixing；Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, obtains final concentration of 10ng/mL Carcinomebryonic antigen solution；The solution is added in centrifuge tube；Then 10mM, Tris-HCl (pH 7.4) buffer solution is used to supplement body Product is incubated 2 hours to 1900 μ L in 37 DEG C of water-baths；
3, by the MoS of commercially available a concentration of 1mg/mL2100 μ L of nanometer sheet solution are added in the solution of step (2), make from The volume of solution is 2mL in heart pipe, is reacted at room temperature 10 minutes after mixing；In centrifuge tube, MoS2Final concentration of the 50 of nanometer sheet μ g/mL, the final concentration of 5nM of aptamers probe；
4, it takes the solution 2mL that step (3) obtains to be added in cuvette, fluorescence spectrometry is carried out, by believing fluorescence Number the detection of intensity realize detection to carcinomebryonic antigen concentration.Cuvette optical length 10mm.Excitation wavelength is 595nm, transmitting Wavelength is 618nm.
Embodiment 1
1, aptamers probe is designed, the synthesis of Hai Sheng works bio-engineering corporation is served；Carcinomebryonic antigen aptamers probe sequence： 5 '-Texas Red (moral gram saxophone is red)-ATACCAGCTTATTCAATT-3 ', obtains aptamers probe powder；Wherein nucleosides Acid sequence comes from document：S.Su,M.Zou,H.Zhao,C.Yuan,Y.Xu,C.Zhang, L.Wang,C.Fan,L.Wang, Shape-controlled gold nanoparticles supported on MoS(2) nanosheets: synergistic effect of thionine and MoS(2)and their application for electrochemical label-free immunosensing,Nanoscale 2015,7,19129-35.
2, by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Centrifuge tube is taken to be added (buffer solution contains the NaCl of a concentration of 100mM, a concentration of 5mM in 998 μ L, 10mM Tris-HCl (pH 7.4) buffer solution KCl and a concentration of 5mM MgCl2, solvent is deionized water), 2 μ L, 5 μM of aptamers probe solutions are added to the centrifugation Guan Zhong shakes mixing；Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, obtains final concentration of 10ng/mL Carcinomebryonic antigen solution；The solution is added in centrifuge tube；Then 10mM, Tris-HCl (pH 7.4) buffer solution is used to supplement body Product is incubated 2 hours to 1800 μ L in 37 DEG C of water-baths；
3, by the MoS of commercially available a concentration of 1mg/mL2200 μ L of nanometer sheet solution are added in the solution of step (2), make from The volume of solution is 2mL in heart pipe, is reacted at room temperature 10 minutes after mixing；In centrifuge tube, MoS2Nanometer sheet it is final concentration of 100 μ g/mL, the final concentration of 5nM of aptamers probe；
4, it takes the solution 2mL that step (3) obtains to be added in cuvette, fluorescence spectrometry is carried out, by believing fluorescence Number the detection of intensity realize detection to carcinomebryonic antigen concentration.Cuvette optical length 10mm.Excitation wavelength is 595nm, transmitting Wavelength is 618nm.
Embodiment 2
1, aptamers probe is designed, the synthesis of Hai Sheng works bio-engineering corporation is served；Carcinomebryonic antigen aptamers probe sequence： 5 '-Texas Red (moral gram saxophone is red)-ATACCAGCTTATTCAATT-3 ', obtains aptamers probe powder；Wherein nucleosides Acid sequence comes from document：S.Su,M.Zou,H.Zhao,C.Yuan,Y.Xu,C.Zhang, L.Wang,C.Fan,L.Wang, Shape-controlled gold nanoparticles supported on MoS(2) nanosheets: synergistic effect of thionine and MoS(2)and their application for electrochemical label-free immunosensing,Nanoscale 2015,7,19129-35.
2, by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Centrifuge tube is taken to be added (buffer solution contains the NaCl of a concentration of 100mM, a concentration of 5mM in 998 μ L, 10mM Tris-HCl (pH 7.4) buffer solution KCl and a concentration of 5mM MgCl2, solvent is deionized water), 2 μ L, 5 μM of aptamers probe solutions are added to the centrifugation Guan Zhong shakes mixing；Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, obtains final concentration of 10ng/mL Carcinomebryonic antigen solution；The solution is added in centrifuge tube；Then 10mM, Tris-HCl (pH 7.4) buffer solution is used to supplement body Product is incubated 2 hours to 1600 μ L in 37 DEG C of water-baths；
3, by the MoS of commercially available a concentration of 1mg/mL2400 μ L of nanometer sheet solution are added in the solution of step (2), make from The volume of solution is 2mL in heart pipe, is reacted at room temperature 10 minutes after mixing；In centrifuge tube, MoS2Nanometer sheet it is final concentration of 200 μ g/mL, the final concentration of 5nM of aptamers probe；
4, it takes the solution 2mL that step (3) obtains to be added in cuvette, fluorescence spectrometry is carried out, by believing fluorescence Number the detection of intensity realize detection to carcinomebryonic antigen concentration.Cuvette optical length 10mm.Excitation wavelength is 595nm, transmitting Wavelength is 618nm.
Embodiment 3
1, aptamers probe is designed, the synthesis of Hai Sheng works bio-engineering corporation is served；Carcinomebryonic antigen aptamers probe sequence： 5 '-Texas Red (moral gram saxophone is red)-ATACCAGCTTATTCAATT-3 '；Its nucleotide sequence comes from document： S.Su,M.Zou,H.Zhao,C.Yuan,Y.Xu,C.Zhang,L.Wang,C.Fan,L. Wang,Shape-controlled gold nanoparticles supported on MoS(2)nanosheets:synergistic effect of thionine and MoS(2)and their application for electrochemical label-free immunosensing,Nanoscale 2015,7,19129-35.
2, by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Centrifuge tube is taken to be added (buffer solution contains the NaCl of a concentration of 100mM, a concentration of 5mM in 998 μ L, 10mM Tris-HCl (pH 7.4) buffer solution KCl and a concentration of 5mM MgCl2, solvent is deionized water), 2 μ L, 5 μM of aptamers probe solutions are added to the centrifugation Guan Zhong shakes mixing；Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl (pH 7.4) buffer solution, obtains final concentration of 10ng/mL Carcinomebryonic antigen solution；The solution is added in centrifuge tube；Then 10mM, Tris-HCl (pH 7.4) buffer solution is used to supplement body Product is incubated 2 hours to 1400 μ L in 37 DEG C of water-baths；
3, by the MoS of commercially available a concentration of 1mg/mL2600 μ L of nanometer sheet solution are added in the solution of step (2), make from The volume of solution is 2mL in heart pipe, is reacted at room temperature 10 minutes after mixing；In centrifuge tube, MoS2Nanometer sheet it is final concentration of 300 μ g/mL, the final concentration of 5nM of aptamers probe；
4, it takes the solution 2mL that step (3) obtains to be added in cuvette, fluorescence spectrometry is carried out, by believing fluorescence Number the detection of intensity realize detection to carcinomebryonic antigen concentration.Cuvette optical length 10mm.Excitation wavelength is 595nm, transmitting Wavelength is 618nm.

Claims (3)

1. one kind being based on two dimension MoS2The biological sensor of nanometer sheet and carcinomebryonic antigen aptamers structure, it is characterised in that：It is It is prepared by following steps,
(1) 5 '-Texas Red-ATACCAGCTTATTCAATT-3 ' of design carcinomebryonic antigen aptamers probe send bioengineering public Department's synthesis, obtains aptamers probe powder；
(2) by the distillation water dissolution of aptamers probe powder, 5 μM of aptamers probe solution is obtained；Take centrifuge tube that 998 μ are added 2 μ L, 5 μM of aptamers probe solutions are added in above-mentioned centrifuge tube by L, 10mM Tris-HCl buffer solutions, shake mixing；By cancer Embryonal antigen is dissolved in 10mM, Tris-HCl buffer solution, obtains the carcinomebryonic antigen solution of known concentration；By the solution be added to it is above-mentioned from In heart pipe, final concentration of 0.1~100ng/mL of carcinomebryonic antigen；Then 10mM, Tris-HCl buffer solution supplement volume are used extremely 1400~1900 μ L, and be incubated 1~3 hour in 37 DEG C of water-baths；
(3) by MoS2Nanometer sheet solution is added in the solution of step (2), makes the volume of solution in centrifuge tube for 2mL, after mixing It reacts 5~20 minutes at room temperature；In centrifuge tube, MoS2Final concentration of 100~300 μ g/mL of nanometer sheet, the end of aptamers probe A concentration of 5nM；
(4) solution that step (3) obtains is added in cuvette, carries out fluorescence spectrometry, then according to the fluorescence measured The numerical relation of intensity and carcinomebryonic antigen concentration is fitted to obtain fluorescence intensity-carcinomebryonic antigen concentration standard curve；To be prepared into To based on two-dimentional MoS2The biological sensor of nanometer sheet and carcinomebryonic antigen aptamers structure.
2. described in claim 1 a kind of based on two dimension MoS2The biological sensing of nanometer sheet and carcinomebryonic antigen aptamers structure Application of the device in detecting carcinomebryonic antigen.
3. as claimed in claim 2 a kind of based on two dimension MoS2The biological of nanometer sheet and carcinomebryonic antigen aptamers structure passes Application of the sensor in detecting carcinomebryonic antigen, it is characterised in that：Carcinomebryonic antigen is dissolved in 10mM, Tris-HCl buffer solution, is obtained The carcinomebryonic antigen solution of unknown concentration；The carcinomebryonic antigen solution of known concentration in claim 1 step (2) is replaced with the solution, It is added in centrifuge tube, carries out the operations described in claim 1 step (2)~(4), then carry out fluorescence spectrometry, The concentration of carcinomebryonic antigen is calculated by standard curve according to the fluorescence intensity measured.
CN201810633861.1A 2018-06-20 2018-06-20 One kind being based on two dimension MoS2Nanometer sheet and carcinomebryonic antigen aptamers structure biological sensor and for detecting carcinomebryonic antigen Pending CN108627646A (en)

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CN114624442A (en) * 2022-05-16 2022-06-14 南京颐兰贝生物科技有限责任公司 Chemiluminescence carcinoembryonic antigen detection reagent based on aptamer signal amplification and preparation method thereof

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CN111721723A (en) * 2019-03-20 2020-09-29 天津大学 Preparation method of hydrogel nano preparation loaded with monitoring probe and medicine
CN111948391A (en) * 2019-05-16 2020-11-17 南京大学 Array sensor based on nano metal organic framework for histological diagnosis of colon cancer
CN111175506A (en) * 2020-02-05 2020-05-19 山西大同大学 Sensor based on hybrid chain reaction and ribozyme and carcinoembryonic antigen detection method
CN111175506B (en) * 2020-02-05 2023-10-27 山西大同大学 Sensor based on hybridization chain reaction and ribozyme and carcinoembryonic antigen detection method
CN111617097A (en) * 2020-06-17 2020-09-04 福州大学 Preparation method and application of [2] -reticular catenane DNA (deoxyribonucleic acid) single-layer array
CN111617097B (en) * 2020-06-17 2021-06-29 福州大学 Preparation method and application of [2] -reticular catenane DNA (deoxyribonucleic acid) single-layer array
CN113390842A (en) * 2021-06-16 2021-09-14 广西医科大学 Aptamer biosensor for rapidly detecting carcinoembryonic antigen in high flux
CN113390842B (en) * 2021-06-16 2024-03-08 广西医科大学 Aptamer biosensor for rapidly detecting carcinoembryonic antigen in high flux
CN114624442A (en) * 2022-05-16 2022-06-14 南京颐兰贝生物科技有限责任公司 Chemiluminescence carcinoembryonic antigen detection reagent based on aptamer signal amplification and preparation method thereof
CN114624442B (en) * 2022-05-16 2022-08-12 南京颐兰贝生物科技有限责任公司 Chemiluminescence carcinoembryonic antigen detection reagent based on aptamer signal amplification and preparation method thereof

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