Common and convenient epitope imprinting method and application of molecular imprinting polymer

An epitope and imprinting technology, applied in chemical instruments and methods, ion exchange, ion exchange regeneration, etc., can solve the problems of inapplicability of molecular imprinting methods, and achieve good universality, versatility, and easy access

Active Publication Date: 2018-11-02
NANJING UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, for non-glycoproteins without cis-dihydroxy groups, the above molecular imprinting method is not applicable due to the absence of boron affinity

Method used

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  • Common and convenient epitope imprinting method and application of molecular imprinting polymer
  • Common and convenient epitope imprinting method and application of molecular imprinting polymer
  • Common and convenient epitope imprinting method and application of molecular imprinting polymer

Examples

Experimental program
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Effect test

preparation example Construction

[0047] The following are the preparation methods of different substrate materials.

[0048] 1. The preparation method when the boric acid functionalized magnetic nanoparticles are used as the base material is as follows:

[0049] Step 1), the preparation method of magnetic nanoparticles can refer to the following methods [Chem. Sci. 2013, 4, 4298-4303; Chem. Eur. J. 2006, 12, 6341-6347];

[0050] Step 2), adding a certain amount of ammonia water and TEOS to the ethanol solution, stirring for a period of time at a suitable temperature, then adding a certain amount of ethanol solution of magnetic nanoparticles, and stirring for a period of time at a suitable temperature to obtain a surface coated silicon magnetic nanoparticles;

[0051] Step 3), dispersing the silicon-coated magnetic nanoparticles obtained in step 2) in an ethanol solution, then adding a certain amount of APTES, and stirring for a period of time at a suitable temperature to obtain amino-functionalized magnetic ...

Embodiment 1

[0080] Example 1: Preparation of glycated epitopes

[0081] Determine the amino acid sequence information of the target protein through protein databases (such as UniProt, Protein Date Bank, etc.). Taking B2M as an example, the C-terminal polypeptide KIVKWDRDM was selected as the epitope sequence, and the polypeptide sequence KIVKWDRDMK with lysine (K) connected to the end of the polypeptide was directly synthesized by solid-phase synthesis, and then the connected K residue was reacted by Schiff base. Combined with fructose (Fru), the fructosylated epitope polypeptide KIVKWDRDMK-Fru is obtained. Take Mb as an example, select its C-terminal polypeptide NYKELGFQG as the epitope sequence, and directly synthesize the polypeptide sequence NYKELGFQGK with K connected to the end of the polypeptide by solid-phase synthesis, and then combine with Fru on the connected K residue through Schiff base reaction, The fructosylated epitope polypeptide NYKELGFQGK-Fru was obtained. Taking CEA ...

Embodiment 2

[0083] Example 2: TEM characterization of glycated epitope-imprinted magnetic nanoparticles

[0084] The morphology and particle size of the C-terminal glycated epitope-imprinted magnetic nanoparticles of B2M were characterized by TEM, and the results are as follows image 3 As shown, the glycated epitope-imprinted magnetic nanoparticles have regular morphology, uniform particle size, and a size of about 150 nm.

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Abstract

The invention discloses a common and convenient epitope imprinting method. The method comprises the following steps: firstly, selecting terminal polypeptide of target protein as a characteristic epitope; carrying out saccharification treatment to obtain saccharified polypeptide and taking the saccharified polypeptide as an imprinting template; anchoring the imprinting template on a boric acid functionalized substrate material through boron affinity interaction; then selecting monomer silylation reagents with different types and ratios and carrying out non-covalent imprinting; accurately imprinting the thickness of an imprinting layer through adjusting the imprinting time, so as to obtain a molecular imprinting polymer. The imprinting technology has the advantages of good commonality, convenience for preparation, good identification performance and the like. A molecular imprinting material prepared by the method can be used for identifying an epitope segment and also can be used for specifically identifying complete target protein, so that the molecular imprinting material has an important application prospect in the fields of separation, enrichment, purification, sensing, proteomics, target identification and the like.

Description

technical field [0001] The invention belongs to the technical field of biomimetic molecular recognition materials and molecular imprinting. Background technique [0002] Antibodies are an important weapon for organisms to resist the invasion of harmful substances such as viruses and microorganisms, and they are also the most widely used biological reagents for biomolecular recognition in the field of life science research. However, antibodies have significant limitations. First, the preparation of antibodies is cumbersome and usually expensive. Secondly, it is necessary to recognize the target molecule as an antigen in the preparation to induce the animal or cell line to produce the corresponding antibody. Therefore, it is difficult to obtain the antibody of the target molecule with some pure products or with poor immunogenicity. Furthermore, antibodies are less stable and reproducible. Therefore, the development of alternatives to antibodies is not only of great scientif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01J20/30B01D15/08
CPCB01D15/08B01J20/268
Inventor 刘震邢荣荣王益佳
Owner NANJING UNIV
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