Mesoporous molecular sieve doped acetylated molecularly imprinted polymer and construction of micro-fluidic chip platform

A mesoporous molecular sieve and molecular imprinting technology, applied in other chemical processes, laboratory containers, chemical instruments and methods, etc., can solve the problems of cumbersome steps in multi-dimensional systems, difficulty in online sample processing, incompatible operating conditions, etc.

Active Publication Date: 2022-07-22
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The multi-dimensional separation system built on the basis of the microfluidic chip has been applied in the field of solid phase extraction, which realizes the online automatic separation and enrichment of samples, and solves the cumbersome steps of the multi-dimensional system. Sample Handling Issues

Method used

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  • Mesoporous molecular sieve doped acetylated molecularly imprinted polymer and construction of micro-fluidic chip platform
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  • Mesoporous molecular sieve doped acetylated molecularly imprinted polymer and construction of micro-fluidic chip platform

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Optimizing the preparation conditions of the molecularly imprinted polymer is to improve its specificity to the template molecule and make the imprinting effect more pronounced. In order to investigate the effect of functional monomer / crosslinker ratio and dopant type on the imprinting effect and adsorption effect of the template. The specific operation steps are as follows:

[0043] a. Preparation method of SBA-15 mesoporous molecular sieve doped acetylated molecularly imprinted polymer:

[0044] The mass fraction of SBA-15 mesoporous molecular sieve (8-12 nm) 0.50%, template KacQLAT (95%, Shanghai Qiangyao Biotechnology Co., Ltd., product number 04010047537) 1.44%, functional monomer zinc acrylate 3.97%, cross-linking agent PEGDA 15.81% and EDMA 9.50%, initiator azobisisobutyronitrile 0.46%, dissolved in a mixed solution of porogen methanol 36.05% and N, N-dimethylformamide 32.27%; ultrasonic (power 150 W) dissolved After 15 min, it was dissolved and clar...

Embodiment 2

[0050] Example 2: Application of the prepared electrochemical sensor based on molecularly imprinted electrode technology in detecting template KacQLAT. Specific steps are as follows:

[0051] a. Preparation of SBA-15@MIP / GCE electrical sensor:

[0052] The SBA-15@MIP / GCE electrical sensor was fabricated using the drop coating method. First, glassy carbon electrodes (GCE) were polished on a polishing cloth with 1.0, 0.3, and 0.05 μM GCE alumina aqueous slurries, respectively, and then washed with sulfuric acid solution (1 M), deionized water, and ethanol for 10 min, respectively. After that, 2 mg of imprinted polymer was added to 1 mL of chitosan solution (0.5% wt, containing 20 μL of acetic acid), followed by 50 μL (1 mg / mL) of MWCNT DMF solution to increase conductivity sex. In order to uniformly disperse the polymer and multi-walled carbon nanotubes, the prepared mixture was sonicated and mixed for 5 min. Finally, 10 μL of the SBA-15@MIP mixture was drop-coated on the su...

Embodiment 3

[0060] Example 3: In order to reduce sample consumption and loss, shorten analysis time and cost, and achieve the goal of automation and integration, a multi-functional microfluidic chip integration platform was built. Specific steps are as follows:

[0061] a. Sample online processing:

[0062] The denaturation chip is connected to protein fractionation before, followed by enzyme reactor and acetylation molecular imprinting enrichment column. The four units are connected in series with thin connecting tubes. The length and size requirements of the denaturation channel and the location distribution of each reservoir are comprehensively considered. The online denaturation column is divided into denaturation I area and denaturation II area. Each area contains multiple groups of serpentine channels, each channel is 2cm long, and the total length of the channel can be adjusted arbitrarily. All channels are 250 μm wide and 100 μm deep. The prepared chip is placed on the jig, and t...

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Abstract

The invention relates to construction of a mesoporous molecular sieve doped acetylated molecularly imprinted polymer and a micro-fluidic chip platform. The imprinted polymer is used as an adsorbent for solid-phase extraction and is used for analyzing acetylation modified peptide. According to the method, the SBA-15 mesoporous molecular sieve is doped into the acetylated molecularly imprinted polymer, so that the imprinting effect of the template peptide can be obviously improved. Meanwhile, polyethylene glycol dimethacrylate is added during preparation and is co-crosslinked with ethylene glycol dimethacrylate, so that the hydrophilicity of the polymer can be improved, non-specific adsorption is effectively reduced, and a remarkable imprinting effect (an imprinting factor 3.2) can be achieved. Then, a plurality of functional areas including protein grading, denaturation, enzymolysis and acetylated peptide enrichment are integrated to construct a micro-fluidic chip platform, and the micro-fluidic chip platform can be used for on-line enrichment of acetylated peptide in a biological sample.

Description

technical field [0001] The invention relates to the construction of a mesoporous molecular sieve doped acetylated molecularly imprinted polymer and a microfluidic chip platform. Background technique [0002] Lysine acetylation (Kac) is a broad dynamic and reversible protein post-translational modification (PTM), which is widely involved in gene transcription, protein degradation, cellular metabolism, stress response, etc. Cellular processes that play an important regulatory role in metabolic-related diseases and neurodegenerative diseases. However, due to the low content of acetylated proteins, their signals will be interfered by a large number of non-acetylated proteins during the identification process, which brings great challenges to their identification and functional studies. In order to better study them, There is an urgent need to develop efficient and specific lysine acetylation enrichment strategies. At present, the commonly used Kac peptide enrichment methods in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01L3/00B01J20/30
CPCB01J20/268B01L3/5027
Inventor 刘照胜张雪黄艳萍
Owner TIANJIN MEDICAL UNIV
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