A nucleic acid aptamer-molecularly imprinted fluorescent sensor for detecting cytochrome c and its preparation method
A nucleic acid aptamer and fluorescent sensor technology, applied in the field of biomedical analytical chemistry, can solve the problems of large size of imprinting template, increased imprinting difficulty, easy to change, etc., and achieves enhanced dual recognition effect, fast reaction speed, and good selection. sexual effect
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[0062] Such as figure 1 As shown, the present invention provides a method for preparing a nucleic acid aptamer-molecularly imprinted fluorescent sensor for detecting cytochrome c, comprising the following steps:
[0063] 1) Preparation of pre-action complex
[0064] Dissolve the template molecule cytochrome c and sulfhydryl-modified cytochrome c nucleic acid aptamer in phosphate buffer, then add methacrylic acid and a cross-linking agent, and react for 12 hours at a temperature of 37°C to prepare the preformed Action complex;
[0065] 2) Preparation of nucleic acid aptamer-quantum dot complex
[0066] Introduce nitrogen into the pre-action compound obtained in step 1) for 20 minutes, remove the air, add double bond-modified manganese-doped zinc sulfide quantum dots and an initiator, and then add phosphate buffer, at a temperature of 37 ° C under nitrogen protection. Under the conditions, react for 12 hours to prepare the nucleic acid aptamer-quantum dot complex;
[0067] 3...
Embodiment 1
[0074] (1) Preparation of manganese-doped zinc sulfide quantum dots modified with mercaptopropionic acid
[0075] (1) Add 5mL 0.1mol / L zinc sulfate solution, 2mL 0.01mol / L manganese chloride solution and 20mL 0.1mol / L 3-mercaptopropionic acid solution into a 50mL three-necked flask, mix the three solutions thoroughly, and then add Stir 23mL deionized water evenly, add 2mol / L sodium hydroxide solution dropwise to the mixed solution to adjust the pH value of the solution to 10;
[0076] (2) Inject 5mL of 0.1mol / L sodium sulfide aqueous solution, pass in nitrogen, and continue stirring for 30min;
[0077] (3) Expose the reaction solution to air atmosphere, heat in a water bath at 50°C, and age for 4 hours;
[0078] (4) The obtained quantum dot solution is precipitated with an equal volume of ethanol, and the supernatant is centrifuged at 12000rpm, and then washed several times with ethanol; the manganese-doped zinc sulfide quantum dots modified by mercaptopropionic acid are disp...
Embodiment 2
[0087] (1) Preparation of manganese-doped zinc sulfide quantum dots modified with mercaptopropionic acid
[0088] In this example, the preparation method of the manganese-doped zinc sulfide quantum dots modified with mercaptopropionic acid is the same as that in Example 1.
[0089] (2) Preparation of double bond-modified Mn-doped ZnS quantum dots
[0090] The preparation method of the above-mentioned double bond-modified manganese-doped zinc sulfide quantum dots in this embodiment is the same as that in Embodiment 1.
[0091] (3) Preparation of nucleic acid aptamer-molecularly imprinted fluorescent sensor (aptamer-MIP / QDs) for detection of cytochrome c
[0092] 1) Dissolve 2.8 nmol of the template molecule cytochrome c and 2.8 nmol of the thiol-modified cytochrome c aptamer in 100 μL of phosphate buffer (pH=7.5, 10 mmol / L), and then add 560 nmol of methacrylic acid (MAA ) and 2mg of N,N'-methylenebisacrylamide (MBA), reacted for 12 hours at a temperature of 37°C to prepare a...
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