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A nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and its preparation method

A technology for nucleic acid aptamers and mycotoxins, which is applied in the field of analytical chemistry and can solve problems such as quantitative and qualitative analysis of target substances, specific identification interference, etc.

Active Publication Date: 2021-06-01
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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  • A nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and its preparation method
  • A nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and its preparation method
  • A nucleic acid aptamer functionalized polymer column for specific recognition of mycotoxins and its preparation method

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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 functional polymer column for specific recognition of mycotoxins and its preparation method. ‑Inorganic hybrid silica gel polymer is used as the matrix, on which the oligomeric silsesquioxane nanoparticles substituted by the sodium salt of acrylic acid are modified, and the anti-mycotoxin nucleic acid aptamers with mercapto groups are modified on the sodium salt nanoparticles, thereby producing obtained affinity column material. The affinity polymerization column prepared in the present invention has a rigid silica gel matrix, and through post-column modification, the surface is rich in mercapto structures and oligomeric sesquipolysiloxane monomers with multiple action sites; through click chemistry, bridge sites can be The anti-mycotoxin nucleic acid aptamer is bonded to form a highly hydrophilic and negatively charged aptamer affinity layer on the surface, and the classification realizes the specific recognition of different mycotoxins.

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 Patents(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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