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A nucleic acid aptamer affinity monolithic column specifically recognizing F2 toxin and its preparation method

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: 2021-06-01
FUZHOU UNIV
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  • Abstract
  • Description
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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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  • A nucleic acid aptamer affinity monolithic column specifically recognizing F2 toxin and its preparation method
  • A nucleic acid aptamer affinity monolithic column specifically recognizing F2 toxin and its preparation method

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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 30min, pass through secondary water until neutral, then pass through 0.1mol / L sodium hydroxide solution for 3h, 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 substances with mercapto groups; then rinse with methanol for 30 minutes, and blow with nitrogen for 3 hours at 70 ° C and 0.4 MPa to obtain capillary tubes with mercapto groups on the inner wall of the ca...

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Abstract

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. Organic-inorganic hybrid silica gel with surface rich in mercapto groups formed by photoinitiated polymerization based on polymercapto-functional monomers, oligomeric polysilsesquipolysiloxane (POSS) and pentaerythritol tetrakis (3-mercaptopropionate) crosslinking agent The polymer is used as the substrate, and the aptamer with thiol groups is bonded to the abundant alkenyl groups on the substrate through the "mercapto-ene" click reaction, and the anti-mycotoxin nucleic acid aptamers with thiol groups are modified on POSS nanoparticles with high coverage density, thereby The obtained monolithic column with affinity for F2 toxin. The monolithic column matrix has the characteristics of a highly cross-linked structure and multiple action sites, and can efficiently immobilize high-density loading of nucleic acid aptamers with specific recognition of F2 toxin on the POSS-based monolithic column. The structure is stable, the method is simple, fast, and available. For efficient and specific recognition and separation of F2 toxin.

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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Patent Type & Authority Patents(China)
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 UNIV
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