Nucleic acid aptamer affinity monolithic column capable of specifically recognizing F2 toxins and preparing method of nucleic acid aptamer affinity monolithic column

A nucleic acid aptamer and monolithic column technology, applied in the field of analytical chemistry, can solve the problems of less research on organic-inorganic hybrid monolithic columns, and achieve the effects of reducing non-specific adsorption and high coverage density

Active Publication Date: 2019-08-09
FUZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the research on nucleic acid aptamer-modified monolithic columns mainly focuses on inorganic silica gel monolithic columns and organic polymeric monolithic columns, while there are few studies on organic-inorganic hybrid monolithic columns.

Method used

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  • Nucleic acid aptamer affinity monolithic column capable of specifically recognizing F2 toxins and preparing method of nucleic acid aptamer affinity monolithic column
  • Nucleic acid aptamer affinity monolithic column capable of specifically recognizing F2 toxins and preparing method of nucleic acid aptamer affinity monolithic column

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Embodiment 1

[0032] A nucleic acid aptamer affinity monolithic column specifically recognizing F2 toxin and the specific steps of its preparation method are as follows:

[0033] (1) Derivatization by thiol on the inner wall of the capillary column

[0034] Pass through the capillary empty column sequentially with 0.1mol / L hydrochloric acid for 30 minutes, pass through secondary water until neutral, then pass through 0.1mol / L sodium hydroxide solution for 3 hours, then pass through again and pass through secondary water until neutral. Inject methanol for 30 minutes, blow dry with nitrogen at 25°C and 0.4MPa; inject a mixture of methanol and mercaptopropyltrimethoxysilane in a volume ratio of 1:1, and react in a constant temperature water bath at 60°C for 24 hours to bond the capillary wall. silanized substance with mercapto groups on it; then rinse with methanol for 30 minutes, blow with nitrogen for 3 hours at 70°C and 0.4MPa, to obtain capillary tubes with mercapto groups on the inner wal...

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Abstract

The invention belongs to the field of analysis chemistry, and particularly relates to a nucleic acid aptamer affinity monolithic column capable of specifically recognizing F2 toxins and a preparing method of the nucleic acid aptamer affinity monolithic column. An organic-inorganic hybrid silica gel polymer which is rich in sulfydryl on the surface and is formed through photo-initiation polymerization of a multi-sulfydryl functional monomer, polyhedral oligomeric silsesquioxanes (POSS), a pentaerythritoltetrakis(3-mercaptopropionate) cross-linking agent and the like is taken as a substrate, anaptamer with sulfydryl is bonded with rich alkenyl on the substrate through a sulfydryl-alkenyl click reaction, an anti-mycotoxin nucleic acid aptamer with sulfydryl is modified on POSS nanometer particles with high coverage density, and thus the monolithic column with an affinity effect on the F2 toxins is prepared. The monolithic column substrate has the advantages of highly-cross-linked structure and multiple acting sites, the nucleic acid aptamer with the function of specific recognition on the F2 toxins can be efficiently loaded and fixed on the POSS-based monolithic column with high density, the structure is stable, the method is simple and quick, and the monolithic column can be used for efficient specific recognition and separation of the F2 toxins.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a nucleic acid aptamer affinity monolithic column for specifically recognizing F2 toxin and a preparation method thereof. Background technique [0002] The monolithic column is a continuous porous monolithic material formed by in-situ polymerization or immobilization in the column, and the surface of the monolithic material can be derivatized according to actual needs. Affinity chromatography is a chromatography method established based on the principle of affinity adsorption and dissociation between biomolecules. The solid-phase carrier is coupled to prepare a specific affinity adsorbent, which is then used as a stationary phase to fill the chromatographic column for the purification of the target substance, showing great potential for development in the field of monolithic column research. [0003] Nucleic acid aptamer is a short-chain DNA or RNA sequence that h...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/30B01J20/285B01J20/28G01N30/60
CPCB01J20/267B01J20/28011B01J20/285G01N30/6052
Inventor 谢增鸿许金花沈浩强林旭聪
Owner FUZHOU UNIVERSITY
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