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Preparation method of phospholipase C sensor based on atom transfer radical polymerization

A technology of atom transfer and phospholipase, applied in instruments, scientific instruments, analytical materials, etc., can solve the problems of complex post-catalyst removal process, catalyst toxicity, catalyst residue, etc., and achieve low cost, high selectivity, and avoid interference Effect

Pending Publication Date: 2022-01-14
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the conventional ATRP method has two disadvantages: first, low-valence transition metal ions are sensitive to air and are inconvenient to use, and an oxygen-free system is often required for polymerization; second, the catalyst has certain toxicity to biological macromolecules such as proteins, and the common The post-treatment process for catalyst removal is complicated, and catalysts often remain
Although electrochemical methods have been successfully applied to the detection of biomarkers due to their advantages of high specificity, high sensitivity, good stability, and convenient operation, however, there has been no related research on electrochemical detection PLC methods based on free radical polymerization. to report

Method used

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  • Preparation method of phospholipase C sensor based on atom transfer radical polymerization
  • Preparation method of phospholipase C sensor based on atom transfer radical polymerization
  • Preparation method of phospholipase C sensor based on atom transfer radical polymerization

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Embodiment 1

[0021] A method of preparing a phospholipase C sensor based on an atomic transfer radical polymerization, and the working electrode is performed in accordance with the following steps:

[0022] Step 1: Place the clean gold electrode in a L-cysteine ​​solution having a concentration of 1 mmol / L, resulting in a modified electrode, the L-cysteine ​​solution is 0.1 mmol / L, pH. = 7.0 PBS formulation;

[0023] Step 2: Place the primary modified electrode in the EDC-NHS solution 5 min, activate the L-cysteine ​​carboxylate end, to obtain a secondary modified electrode, the EDC-NHS solution is 0.1 mmol / L, pH = 7.0 The PBS is formulated, the EDC content is 20 mg / ml, the NHS content is 4 mg / ml;

[0024] Step 3: Place the secondary modified electrode for 6 hours in the phosphatidyllet alkanlamine solution, then soaked the electrode for a 2 mmol / l bovine serum albumin (BSA), and the non-specific binding site on the gold electrode is closed. Three modified electrodes, the phospholi...

Embodiment 2

[0034] A method of preparing a phospholipase C sensor based on an atomic transfer radical polymerization, and the working electrode is performed in accordance with the following steps:

[0035] Step 1: Place the clean gold electrode in a L-cysteine ​​solution having a concentration of 0.1 mmol / L, resulting in a modified electrode, and the L-cysteine ​​solution is 0.1 mmol / L, pH. = 7.0 PBS formulation;

[0036] Step 2: Place the primary modified electrode in an EDC-NHS solution for 30 min, activate the L-cysteine ​​carboxylate end, to obtain a secondary modified electrode, the EDC-NHS solution is 0.1 mmol / L, pH = 7.0 The PBS is formulated, the EDC content is 5 mg / ml, the NHS content is 1 mg / ml;

[0037] Step 3: Further, the secondary modified electrode is placed in a phosphatidyllet amine solution for 3 hours, and then the electrode is soaked in a 2 mmol / L bovine serum albumin (BSA) for 10 min, and the non-specific binding site on the gold electrode is obtained. Three m...

Embodiment 3

[0043] A method of preparing a phospholipase C sensor based on an atomic transfer radical polymerization, and the working electrode is performed in accordance with the following steps:

[0044] Step 1: Place the clean gold electrode for 12 hours in the L-cysteine ​​solution having a concentration of 0.2 mmol / L, resulting in a modified electrode, the L-cysteine ​​solution is 0.1 mmol / L, pH. = 7.0 PBS formulation;

[0045] Step 2: Place the primary modified electrode for 10 min, activate the L-cysteine ​​carboxylate end, to obtain a secondary modified electrode, the EDC-NHS solution is 0.1 mmol / L, pH = 7.0 PBS is formulated, the EDC content is 10 mg / ml, NHS content is 2 mg / ml;

[0046] Step 3: The secondary modified electrode is placed in a phosphatidyllet alkanlamine solution for 1 hour, and then the electrode is soaked with a 2 mmol / l bovine serum albumin (BSA) for 10 min, and the non-specific binding site on the gold electrode is obtained. Three-time modified electrod...

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Abstract

The invention discloses a preparation method of a phospholipase C sensor based on atom transfer radical polymerization. A polymethylacrolein modified electrode as a working electrode is prepared from a specific ATRP (Atom Transfer Radical Polymerization) polymer solution under an illumination condition, and then silver nanoparticles are effectively introduced into an MVL ATRP molecular chain for signal amplification of the working electrode of the electrochemical sensor. The prepared electrochemical biosensor is very sensitive in PLC detection, the linear detection range of phospholipase C concentration is 1-106 mU / L, and the detection limit is 0.1032 mU / L (S / N = 3). Compared with the prior art, the method has the advantages of high sensitivity, strong operability and the like. In addition, the method is low in background signal, high in selectivity, capable of avoiding interference of false positive results, low in cost and high in benefit.

Description

Technical field [0001] A phospholipase C sensor preparation method is disclosed, in particular, a phospholipase C sensor preparation method based on atomic transfer radical polymerization. Background technique [0002] Enzymes are an extremely important bio-catalyst, which can be efficient and specific in the biological body, and gradually become the focus of biosensor research and development. Phosphatase C (PLC) is a phosphate diesterase which can catalyze phosphate bond hydrolysis to produce diacyl glycerol (DAG) and phosphate groups (such as phosphorylation choline and phosphorylation inositol), in membrane transport, cellular mechanism regulation , Both digestive metabolism has biological functions. The hydrolysis reaction of PLC is often related to abnormal choline metabolism in cancer cells. Different types of PLCs are associated with human cancers, immune disorders and neurodegenerative diseases. At present, fluorescence, acid-base titration, nuclear magnetic resonance, r...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48C08F116/34
CPCG01N27/327G01N27/3277G01N27/3278G01N27/48C08F116/34C08F2438/01
Inventor 孙越李希雯那杉杉王晶晶宋玮伍丽媛赵洹影
Owner LIAONING NORMAL UNIVERSITY
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