Nano fluorescence biosensor as well as preparation method and application thereof
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A biosensor and nano-fluorescence technology, applied in the field of biochemistry and life science analysis, to achieve the effect of strong controllability, sensitive fluorescence signal, and simple structure
Active Publication Date: 2016-07-20
QINGDAO UNIV OF SCI & TECH
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Embodiment 1
[0037] A preparation method for preparing the nano fluorescent biosensor of the present invention, comprising the steps of:
[0038] (1) Add 10 μL of 1.0×10 -5 MDAS1 and 10 μL 1.0 x 10 -3Dilute MDTT solution to 100 μL with PBS, shake overnight at room temperature, and modify DNAS1 onto the surface of gold nanocages;
[0039] (2) Magnetically separate the above solution, wash with PBS buffer solution and add 100 μL of 1.0×10 -5 M Rhodamine B solution, shaking overnight at room temperature;
[0040] (3) Mix 200 μL gold nanoparticles with 10 μL 1.0×10 -5 MDAS2 and 10 μL 1.0 x 10 -3 The mixed solution of MDTT was shaken overnight at room temperature;
[0041] (4) Mix the solution obtained in step (3) with the solution obtained in step (2), react at room temperature for 2 hours, and the hybridization reaction of DNAS2 and DNAS1 occurs, so that the gold nanoparticles are assembled on the surface of the gold nanocage to form a gold nanocage Pore plugged caps to prevent Rhodam...
Embodiment 2
[0044] A kind of detection that utilizes nano fluorescent biosensor of the present invention to be used for ATP, method is as follows:
[0045] (1) Add 10 μL ATP sample solution to the ATP-aptamer complex, dilute to 100 μL with PBS, shake at 37°C for 1 hour, ATP specifically binds to the aptamer, and the primer DNAS3 is competitively removed by a competitive displacement reaction;
[0046] (2) Magnetically separate the above solution, take the supernatant and add it to the PBS suspension of the nano-fluorescent biosensor of the present invention, react at 37°C for 2h, hybridize between primers S3 and S2, add exonuclease ExoⅢ, and react at 37°C 2h, the enzyme digestion reaction cuts S2 in the double strand, the primer S3 enters the cycle, the hole cap switch is continuously opened, and the fluorescent molecules are released;
[0047] (3) Magnetically separate the above solution, collect the supernatant, and detect its fluorescence signal, fluorescence detection conditions: exci...
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Abstract
The invention provides a new nano fluorescence biosensor with a biological release control function as well as a preparation method and application thereof, wherein gold nano particles are used as an orifice cap of a gold nano cage. The sensor has the following characteristics: a DNA hybridization technology is adopted, and the gold nano particles are assembled on the surface of the gold nano cage to form an orifice cap switch to realize blocking of matters in the gold nano cage. The nano biosensor provided by the invention has the characteristics of simple structure, sensitive signal, strong controllability and the like. By adopting the nano fluorescence biosensor provided by the invention, high-sensitivity detection of ATP can be realized in a range of 1.0*10<-9>-1.0*10<-7>M; and the nano fluorescence biosensor has the advantages of sensitivity, economy, easiness in operation and the like, gains relatively great application potential and broad application prospects in the fields such as biomedicine and life science, and provides a new way and method for early diagnosis and treatment of serious diseases and particularly for the study and application of tumor-targeted drugs and photo-thermal therapy based on the nano carrier release control technology.
Description
technical field [0001] The invention belongs to the field of biochemistry and life science analysis, and in particular relates to a nanometer fluorescent biosensor and its preparation method and application. Background technique [0002] In recent years, nanotechnology has been extensively studied and applied in the fields of biomedicine and life sciences, among which gold nanocages, as an emerging gold nanomaterial, have attracted much attention. The nanomaterial is a hollow and porous cage-like particle structure, and small holes are distributed on the smooth cage wall surface. Compared with traditional spherical gold nanoparticles, the localized surface plasmon resonance peak of gold nanocages is located in the near-infrared region of 700-900nm, which is of great significance for biomedicine, especially for the study of living tissues and cells. The detection technology of bioactive molecules that combines gold nanocages with gold nanoparticle pore caps and enzyme digest...
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