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ZnTCPP@MOF-based electrochemical immunoassay method for microcystic toxins

A microcystin and immunoassay technology, applied in the field of electrochemical sensing, can solve the problems of low detection limit, narrow linear range and high cost, and achieve the effects of convenient operation, high detection sensitivity and low cost

Active Publication Date: 2017-10-24
迈科若(苏州)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problems of high cost, narrow linear range and low detection limit in existing microcystin detection methods, the present invention provides a simple, fast and low-cost electrochemical immunoassay microcapsule based on ZnTCPP@MOF Algal toxin method

Method used

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  • ZnTCPP@MOF-based electrochemical immunoassay method for microcystic toxins
  • ZnTCPP@MOF-based electrochemical immunoassay method for microcystic toxins
  • ZnTCPP@MOF-based electrochemical immunoassay method for microcystic toxins

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Experimental program
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Effect test

Embodiment 1

[0026] The preparation method of porphyrin functionalized metal-organic framework material ZnTCPP@MOF can be obtained by referring to the literature [Porphyrin-Encapsulated Metal-Organic Frameworks as Mimetic Catalysts for Electrochemical DNA Sensing via Allosteric Switch of Hairpin DNA]. The specific steps are as follows:

[0027] (1) 0.5g trimesic acid (H 3 BTC) and 30mg zinc porphyrin (ZnTCPP), added to 15ml 1:1 (V:V) mixture of absolute ethanol and DMF, and then mixed with 7.5ml Cu(NO 3 ) 2 ·3H 2 O (1.04g) aqueous solution was mixed, and the above-mentioned mixed solution was ultrasonicated for 20 minutes to make it fully homogeneously dispersed;

[0028] (2) Divide the mixed liquid into 50ml polytetrafluoroethylene tanks and seal them with a high-pressure reactor, put the reactor into an electric heating constant temperature blast drying oven, heat at 60°C for 7 hours, and then cool to room temperature.

[0029] (3) Centrifuge the cooling mixture in step (2) at a speed...

Embodiment 2

[0032]Dissolve 1 mg, 5 mg, 10 mg, 15 mg and 20 mg of TOAB in 1 mL of 10 mg / mL ZnTCPP@MOF toluene solution, respectively, take 20 μL of the above solution to modify on the glassy carbon electrode, place it to dry at room temperature, and use the platinum wire electrode as the counter electrode , the Ag / AgCl electrode is used as the reference electrode, and the above-mentioned modified electrode is used as the working electrode to contain 0.1M KNO 3 The 0.1M pH7.4PBS is used as the electrolyte, and the ECL signal intensity is detected. Electrochemiluminescence system parameter setting: bias voltage of photomultiplier tube is 1kV, amplification stage is 3, scanning potential range is set to -1.7~-0.5V, and scanning rate is 100mV·s-1. Test results such as Figure 4 As shown in (A), when the amount of TOAB is too small, the stability of ZnTCPP@MOF on the electrode is not enough and it is easy to fall off. When the amount of TOAB is too large, the film of TOAB on the surface of the...

Embodiment 3

[0034] The assembly process of MC-LR electrochemical immunosensor is as follows: Figure 5 As shown, the main steps are as follows:

[0035] 20 μL of 10 mg / mL ZnTCPP@MOF and 10 mg / mL tetraoctylammonium bromide (TOAB) toluene mixed solution was dropped to modify the surface of GCE electrode, and dried naturally at room temperature to prepare ZnTCPP@MOF(TOAB) / GCE electrode; Add 20 μL of a mixed aqueous solution of 400 mM EDC and 100 mM NHS to the ZnTCPP@MOF(TOAB) / GCE electrode dropwise to activate the carboxyl group of the metal-organic framework ZnTCPP@MOF. After reacting at room temperature for 30 minutes, rinse with 0.01M PBS solution of pH 7.4 On the electrode surface, modify 20 μL antigen MC-LR on the electrode, and incubate in a 37°C incubator for 1 hour, then rinse the electrode surface with PBS solution to remove excess MC-LR, and then prepare MC-LR / ZnTCPP@MOF (TOAB) / GCE; Finally, 20 μL of 10 μg / mL anti-MC-LR monoclonal antibody (anti-MC-LR) was added dropwise on the mo...

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Abstract

The invention discloses a ZnTCPP@MOF-based electrochemical immunoassay method for microcystic toxins. The method comprises the following steps: modifying an electrode by a porphyrin-functionalized metal-organic framework ZnTCPP@MOF; increasing the stability of the ZnTCPP@MOF on the electrode by utilizing tetra-n-octylammonium bromide; and dripping microcystic toxin MC-LR antigen and monoclonal antibody, and assembling to form an electrochemical immunosensor. The electrochemical immunosensor prepared by the method disclosed by the invention is capable of realizing high-sensitivity and high-specificity detection of the microcystic toxin MC-LR, the generated electrochemical luminescence signal is stable, the lower limit of detection on the microcystic toxin MC-LR is low, and the method has excellent application prospects in the field of detection of biological and environmental microcystic toxins.

Description

Technical field [0001] The invention belongs to the field of electrochemical sensing technology, and specifically relates to a ZnTCPP@MOF-based electrochemical immune detection method for microcystin. Background technology [0002] For a long time, eutrophication of water bodies and cyanobacterial blooms have occurred frequently, resulting in increasingly serious pollution of microcystins (MCs) in water bodies. Microcystin MC-LR is extremely harmful to aquatic organisms and human health. The World Health Organization has stipulated that the safety limit of MC-LR in drinking water is 1.0 μg / L, therefore the monitoring of MC-LR concentration in drinking water sources is particularly important. [0003] Zhang Xinai and others modified the carbon nanotube nano-gold composite film on the surface of the glassy carbon electrode, and used the specific recognition between MC-LR and antibodies to construct an immunoassay mode with a "sandwich" structure. The linear detection range o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N21/76
Inventor 邓盛元宋宏鑫姚传广郑晨昱崔宏达
Owner 迈科若(苏州)医疗科技有限公司
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