Immunofluorescent chromatographic test strip for copper ion detection and application thereof
An immunofluorescence and chromatography test paper technology, applied in measurement devices, analytical materials, instruments, etc., can solve the problems of difficult to adapt to environmental pollution events, environmental quality screening and detection, long analysis time, low efficiency, etc., to achieve rapid and efficient detection, High sensitivity and easy to use effect
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Embodiment 1
[0052] The preparation of embodiment 1 immunofluorescence chromatography test strip
[0053] Preparation and Identification of Complete Antigen for Copper Ion Detection
[0054] Coupling of Cu ions to the carrier protein OVA via the bifunctional chelator p-SCN-Bn-NOTA
[0055] Prepare pH8.9, 0.1M HEPES buffer solution, dissolve OVA protein powder in HEPES buffer solution to obtain OVA protein solution with a concentration of 10mg / ml; dissolve NOTA powder in DMSO solution to obtain NOTA with a concentration of 25mg / ml solution; add the NOTA solution dropwise to the OVA protein solution, stir and chelate in the ice box for 3 hours, and prepare the NOTA-OVA solution overnight at 4°C (the molar ratio of NOTA to OVA is about 70:1);
[0056] CuSO 4 ·5H 2 O powder was dissolved in ultrapure water to obtain a copper ion solution with a concentration of 1.0 mg / ml; the Cu 2+ The solution was added dropwise to the NOTA-OVA solution (Cu 2+ The molar ratio of Cu-NOTA-OVA to NOTA is ab...
Embodiment 2
[0067] The performance evaluation of the copper ion concentration in the detection sample with the immunofluorescence chromatography test strip in embodiment 1
[0068] Preparation of the standard curve: add a series of gradient concentrations (0, 0.8, 1.6, 3.12, 6.25, 12.5, 25, 50, 100ng / ml) competitor standard substance Cu-NOTA dropwise to 75ul per card. After 15 minutes, read the fluorescent signal on the machine. For each detection, 10 parallel groups were used to obtain the mean value, and the standard curve and inhibition rate (sensitivity) were calculated. Set the fluorescent signal T / C value of 0ng / ml standard well as B 0 , the T / C value of other wells is B, to detect the inhibition rate (B 0 -B) / B 0 ×100% is the ordinate, with Cu2+ Concentration log10 logarithm as the abscissa, draw the standard inhibition curve.
[0069] The photo after detecting copper ion in described immunofluorescence chromatography test strip product standard curve preparation process is as ...
Embodiment 3
[0097] Utilize the immunofluorescence chromatography test strip described in embodiment 1 to detect copper ion concentration in Chinese herbal medicine sample, soil sample, and water sample
[0098] 1. Microwave digestion recovery rate of Chinese herbal medicine and soil samples
[0099] Conditions of microwave digestion Accurately weigh 0.2000g of the sample, add 8ml of nitric acid, and pre-digest in a fume hood overnight or preheat the acid catcher at 130°C for 30 minutes; select the digestion program (climb at 130°C for 10 minutes, hold at 130°C for 3 minutes; ℃ for 5 minutes; 180 ℃ for 3 minutes, 180 ℃ for 40 minutes; cooling); the specific operation of coupling with the chelating agent after the microwave digestion of the sample described in the present invention: after the Chinese herbal medicine sample is digested, use a 10ml volumetric flask to make up the volume, and take 5ml for digestion Add 1-2 μl of 5 μg / μl single NOTA solution to the solution, mix and react for 1...
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