Nucleic acid aptamer functionalized polymerization column for specific recognition of mycotoxin and preparation method of column

A nucleic acid aptamer and mycotoxin technology, applied in the field of analytical chemistry, can solve problems affecting quantitative and qualitative analysis of target substances, specific identification interference, etc., to achieve high-density loading and reduce non-specific adsorption.

Active Publication Date: 2019-08-13
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the two major factors of hydrophobic interaction and non-specific adsorption caused by silanol, non-specific adsorption will cause certain interference to the specific recognition of analytes and affect the quantitative and qualitative analysis of the target.

Method used

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  • Nucleic acid aptamer functionalized polymerization column for specific recognition of mycotoxin and preparation method of column
  • Nucleic acid aptamer functionalized polymerization column for specific recognition of mycotoxin and preparation method of column
  • Nucleic acid aptamer functionalized polymerization column for specific recognition of mycotoxin and preparation method of column

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Embodiment 1

[0039] A nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and its preparation method. The specific steps are as follows:

[0040] (1) Preparation of organic-inorganic hybrid silica gel matrix

[0041] Weigh 880 mg of polyethylene glycol (PEG-10000) according to the ratio in Table 1, and dissolve 900 mg of urea in 25 mL of 0.01mol / L acetic acid aqueous solution, and add the volume ratio (v / v) at 3:1~5 : 1 mixture of tetramethoxysilane (TMOS) and 3-mercaptopropyltriethoxysilane (MPTMS) was stirred in an ice-water bath for 45 minutes to obtain a sol solution; the prepared solution was placed in a centrifuge tube Mix well, degassed by ultrasonic, inject into quartz capillary, seal both ends and place in 50 ℃ water bath for constant temperature reaction for 16 h; take out the prepared monolithic column, connect to liquid chromatograph with high-pressure solvent pump, Under the state of 11.0 Mpa, methanol was used as the mobile phase to wash...

Embodiment 2

[0054] Select formula C in Table 1 to prepare blank columns of unmodified nucleic acid aptamers, respectively modify the anti-ochratoxin A nucleic acid aptamer, anti-zearalenone nucleic acid aptamer, and anti-aflatoxin B1 nucleic acid aptamer The affinity polymerization column of the body, the column length is 5cm, and the equilibration, sample loading, washing and elution are carried out respectively. The specific steps are as follows: (1) Equilibration: first equilibrate with pH 8.50 buffer for 30min, and the flow rate is 0.10 mL / min. Pressure 500psi, described buffer solution is by 10mmol / L Tris-HCl, 120 mmol / LNaCl, 5 mmol / L KCl and 20mmol / L CaCl 2 Composition; (2) Loading: Inject 20 μL of 10 ng / mL ochratoxin A (OTA), zearalenone (ZEN), and aflatoxin B respectively 1 (AFB 1 ) solution to be enriched on the monolithic column for 1 hour, the injection conditions are flow rate 0.05 mL / min, pressure 500psi; (3) Cleaning: install the enrichment column on the liquid chromatograp...

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Abstract

The invention discloses a nucleic acid aptamer functionalized polymerization column for specific recognition of mycotoxin and a preparation method of the column. The polymerization column is an affinity column material prepared by adopting an organic-inorganic hybrid silica gel polymer which is formed by polycondensation of organic siloxane gel and contains abundant sulfhydryl on the surface to serve as a matrix, modifying oligomeric silsesquioxane nanoparticles substituted by sodium acrylate on the basis of the matrix and then modifying anti-mycotoxin nucleic acid aptamers with sulfhydryl onsodium salt nanoparticles. The affinity polymerization column prepared by means of the method is provided with the rigid silica gel matrix; through post-modification of the column, abundant sulfhydrylstructures and multiple acting sites of oligomeric polysilsesquioxane monomers are formed on the surface. The anti-mycotoxin nucleic acid aptamers can be bonded to bridging sites through click chemistry to form aptamer affinity interaction layers with high hydrophilicity and negative charges on the surfaces, and the specific identification of different mycotoxins is realized by class.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and a preparation method thereof. Background technique [0002] Selective identification of targets is one of the key technologies to achieve accurate analysis of complex samples. Aptamer-functionalized affinity monolithic columns have the advantages of fast mass transfer resistance, good affinity, and easy preparation. As an emerging affinity technology, they are widely used in the selective extraction, separation, and detection of target analytes in complex samples. However, due to the two major factors of hydrophobic interaction and non-specific adsorption caused by silanol, non-specific adsorption will cause certain interference to the specific recognition of analyte, and affect the quantitative and qualitative analysis of the target. The ideal nucleic acid aptamer affi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/285B01D15/38G01N30/02G01N33/543G01N33/544G01N33/551B01J20/30
CPCB01D15/3819B01J20/285G01N30/02G01N33/54346G01N33/54393G01N33/544G01N33/551
Inventor 林旭聪沈浩强朱丹丹谢增鸿
Owner FUZHOU UNIV
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