Molecularly-imprinted monolithic column modified and enriched through benzoylation as well as preparation method and application of molecularly-imprinted monolithic column

A technology of molecular imprinting and benzoylation, which is applied in separation methods, chemical instruments and methods, and other chemical processes, and can solve problems such as weak force, interference, and complexity of MIPs

Active Publication Date: 2021-09-10
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Due to the complex sample systems such as proteins and peptides, the interference is serious, and the MIPs prepared only with DESs monomers as functional monomers in the imprinting of proteins and peptides not only have weak force, but also cannot fix enough templates to form effective recognition sites. And there is a serious problem of non-specific adsorption

Method used

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  • Molecularly-imprinted monolithic column modified and enriched through benzoylation as well as preparation method and application of molecularly-imprinted monolithic column
  • Molecularly-imprinted monolithic column modified and enriched through benzoylation as well as preparation method and application of molecularly-imprinted monolithic column
  • Molecularly-imprinted monolithic column modified and enriched through benzoylation as well as preparation method and application of molecularly-imprinted monolithic column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Evaluation of the recovery rate of the benzoylated peptide SGRGKbz on the molecularly imprinted monolithic column enriched by benzoylation modification prepared in the presence of monofunctional and bifunctional monomers.

[0034] First, molecularly imprinted monolithic columns and non-imprinted monolithic columns enriched by benzoylation modification were synthesized by in situ thermal polymerization. The recovery rate of the imprinted monolithic column to SGRGKbz was evaluated under appropriate solid-phase extraction conditions and high-performance liquid phase conditions. The synthesis reaction conditions and treatment methods were as follows:

[0035] Synthesis of benzoylated-enriched molecularly imprinted monolithic columns containing bifunctional monomers by in situ thermal polymerization:

[0036] Mix choline chloride and methacrylic acid evenly at a molar ratio of 1:2, heat in an oil bath at 90°C for 1 hour, and obtain a uniform and transparent solution that is ...

Embodiment 2

[0043] In order to clarify the effect of the content ratio of zinc acrylate and DESs on the recovery rate of SGRGKbz enriched by molecularly imprinted monolithic column solid phase extraction with benzoylation modification, benzoylation modification enrichment at different contents of zinc acrylate and DESs was prepared Molecularly imprinted monolithic columns. The specific operation steps are as follows:

[0044] a. Synthesize molecularly imprinted monolithic columns and non-imprinted monolithic columns containing different contents of zinc acrylate and DESs with different contents of benzoylation modification and enrichment by the same method as above (Example 1). Except for the addition of functional monomers with different ratios of zinc acrylate and DESs, the rest are the same. The mass ratios of zinc acrylate to DESs were 1.061, 0.68, 0.454, and 0.303, respectively.

[0045] b. The same method as above (Example 1) was used to carry out solid phase extraction of SGRGKbz...

Embodiment 3

[0048] In order to clarify the effect of different template and functional monomer content on the recovery rate of SGRGKbz enriched by molecularly imprinted monolithic column solid phase extraction of benzoylation modification, the benzoylation modification enrichment method under different template and functional monomer content was prepared. Molecularly imprinted monolithic columns. The specific operation steps are as follows:

[0049] a. Synthesize molecularly imprinted monolithic columns and non-imprinted monolithic columns containing different templates and functional monomer contents enriched by benzoylation by the same method as above (Example 1). In addition to adding different amounts of templates, the contents of the fixed functional monomers and other substances are kept constant so as to change the ratio of templates to functional monomers. The mass ratios of templates and functional monomers were 0.24, 0.29, and 0.49, respectively.

[0050] b. The same method as...

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Abstract

The invention relates to a molecularly-imprinted monolithic column modified and enriched through benzoylation as well as preparation method and application of the molecularly-imprinted monolithic column. According to the molecularly-imprinted monolithic column modified and enriched through benzoylation, a synthesized benzoylation-modified peptide SGRGKbz is used as a template, zinc acrylate and DESs monomers (choline chloride-methacrylic acid and ChCl-MAA) are selected as binary functional monomers, and the imprinted monolithic column which is high in recognition capacity, high in selectivity and high in affinity is prepared. The imprinted monolithic column prepared by using the method disclosed by the invention has a high recovery rate of SGRGKbz, and the recovery rate can reach 97.3%; and 12 benzoylation-modified peptide fragments can be detected after mouse liver protein enzymatic hydrolysate is enriched by the imprinted monolithic column, and only one benzoylation modified peptide fragment is detected when the mouse liver protein enzymatic hydrolysate is not enriched. The imprinted monolithic column prepared by the invention can realize specific enrichment of benzoylation-modified peptides, provides a new choice for benzoylation modification and enrichment, and has very important significance for research of benzoylation modification proteomics.

Description

technical field [0001] The invention relates to a molecularly imprinted monolithic column for benzoylation modification and enrichment, a preparation method and application thereof. Background technique [0002] Protein post-translational modification (PTMs) refers to a complex chemical modification process experienced by proteins after translation. Several amino acid residues are modified with specific chemical groups, or the properties of proteins are changed by enzymatic cleavage by proteolytic enzymes. . After post-translational modification, the protein structure is more complex, the molecular regulation is more refined, and its biological function is more perfect. Common post-translational modifications include phosphorylation, acetylation, methylation, ubiquitination, glycosylation, etc. In recent years, thanks to the development of biological mass spectrometry technology, some new post-translational modifications have been reported successively, such as succinylati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F222/14C08F220/06C08F220/34B01J20/26B01J20/30B01D15/20
CPCC08F222/102B01J20/268B01D15/20C08F220/06C08F220/34
Inventor 刘照胜蒲婉蓉黄艳萍
Owner TIANJIN MEDICAL UNIV
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