A kind of preparation method of L-cysteine self-powered biosensor
A biosensor, cysteine technology, applied in instruments, scientific instruments, measuring devices, etc., can solve problems such as difficulty in meeting accurate, simple, fast, and highly sensitive detection requirements, high analysis costs, and dependence on large-scale instruments, etc. Achieve the effect of improving anti-interference ability, improving detection sensitivity and avoiding reaction
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[0028] Step 1. Preparation of N-GR: Dissolve 30~60 mg graphene oxide (GO) in 30~50 mL deionized water (pH10, adjust pH with 30% ammonia water), then add 2~5 mL hydrazine hydrate, stir After 10-30 min, heat at 200 °C for 3-5 h, wash with ultrapure water, and dry at 60-80 °C for 8 h under vacuum; then add 2-5 mL of PDDA, and sonicate for 0.5-1 h. washing and drying;
[0029] Step 2. Preparation of N-GR / AuNPs: Mix 30-50 μL of prepared N-GR with 5-10 mL of 10-100 nM AuNPs, shake gently overnight at room temperature, and centrifuge (8000-12000, 5- 20 min) to remove excess AuNPs to obtain N-GR / AuNPs;
[0030] Step 3. Preparation of DNA2-GOD: Mix 50~100 μL of 1~5 mM DNA2 with 2~5 mL of 5~10 mg / mL EDC / NHS solution at room temperature for 1~2 h, then wash with phosphate buffer solution ( PBS) for 8-10 times of purification; mix 400-600 μL of 5-10 mg / mL GOD with 1-2 mg of 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinate Mix imide ester sodium salt (sulfo-SMCC) for 1-...
Embodiment 1
[0048] By capturing Ag + Immobilized conjugates to construct self-powered biosensors for the detection of L-Cys:
[0049] (1) Preparation of N-GR / AuNPs: Dissolve 60 mg GO in 30 mL deionized water (pH 10, adjust pH with 30% ammonia water), then add 2 mL hydrazine hydrate, stir for 10 min, Heated for 3 h, washed with ultrapure water, and dried under vacuum at 80 °C for 8 h; 5 mg of the above-mentioned N-GR was dispersed in 5 mL of 1% PDDA saline solution, dissolved by ultrasonication for 1 h, and the above The solution was centrifuged to remove the remaining PDDA (8000, 10 min) to obtain PDDA-modified N-GR; the above precipitate was mixed with 5 mL of AuNPs at a concentration of 10 nM, shaken gently at room temperature overnight, and centrifuged (8000, 10 min ) to remove excess AuNPs, and the final product was redispersed to a concentration of 1 mgmL -1 N-GR / AuNPs;
[0050] (2) Preparation of DNA2-GOD: Mix 60 μL of 1 mM DNA2 with 2 mL of 10 mg / mL EDC / NHS solution at room tem...
Embodiment 2
[0061] by trapping Hg 2+ Immobilized conjugates to construct self-powered biosensors for the detection of L-Cys
[0062] (6) DNA2-GOD bioconjugate NG-AuNPs / Hg 2+ Preparation of / MCH / DNA1 / UPCS / AuNPs bioanode: Take 50 μL UPCS / AuNPs drop-coated on the surface of carbon cloth electrode, dry at 37 ℃ for 2 h, then immerse the electrode in 10 mg / mL EDC / NHS After 0.5 h in the solution, the excess EDC / NHS solution was rinsed with ultrapure water; 75 μL of 1 mM DNA1 was applied onto the surface of the above-mentioned activated electrode, stored at room temperature for 12 h, and then 20 μL of 1 mM MCH was added to react 0.5 h, to block non-specific binding sites, and centrifuge to remove excess MCH; when 15 μM Hg 2+ When present, 50 μL DNA2-GOD conjugate N-GR / AuNPs solution was drop-coated on the electrode surface, and reacted at 37 °C for 1.5 h to prepare DNA2-GODbioconjugate NG-AuNPs / Hg 2+ / MCH / DNA1 / UPCS / AuNPs bioanode, stored at 4 ℃ for future use;
[0063] DNA1 aptamer sequence: ...
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