Preparation method of bovine serum albumin molecular surface imprinted polymer microspheres

Abstract

The invention provides a preparation method of bovine serum albumin molecular surface imprinted polymer microspheres. The method comprises the following steps: 1, adding methacryloyl chloride into an acetone solution containing crosslinked polyvinyl alcohol microspheres to form modified microspheres; 2, combining acryloyloxyethyl trimethyl ammonium chloride with bovine serum albumin molecules in a neutral aqueous solution by virtue of strong electrostatic interactions so as to form a host-guest complex; 3, carrying out a crosslinking copolymerization reaction on the metacrylate modified microspheres, the host-guest complex, a crosslinking agent N,N'-methylene bisacrylamide and an initiating agent so as to achieve the surface imprinting of the bovine serum albumin molecules; and 4, repeatedly flushing the imprinted microspheres with sodium chloride and then flushing the imprinted microspheres with distilled water so as to obtain the bovine serum albumin molecular surface imprinted polymer microspheres with a large quantity of template molecular imprinted cavities retained on the surfaces. According to the method, the protein imprinting is carried out on the basis of the electrostatic interactions between the template protein and each functional monomer; and a new molecular design thought for developing a protein molecular imprinting technology is provided. Thus, the method provided by the invention has a positive reference value in the protein separation and purification field.

Description

technical field [0001] The invention belongs to the technical field of protein engineering and functional macromolecules, and relates to the preparation of molecular surface imprinted polymer microspheres with the characteristics of identifying and selecting biological macromolecules, in particular to the preparation of bovine serum albumin molecular surface imprinted polymer microspheres method. Background technique [0002] Molecularly imprinted polymers (MIPs) are well-tailored functional polymer materials, in which a large number of imprinted holes of template molecules are distributed inside. and other aspects are highly matched with template molecules, so that molecularly imprinted polymers can specifically recognize template molecules at the molecular level, which is similar to the specificity of antibody-antigen recognition and binding, so it is called artificial Antibody (Artificial antibody) or artificial acceptor (Artificial acceptor). Molecular imprinting techn...

AI Technical Summary

Problems solved by technology

[0003] At present, there are obviously several problems in western blotting: (1) The traditional embedding blotting method is not suitable for protein blotting

In the highly cross-linked block polymer network, the mobility of protein macromolecules is poor, resulting in low imprinting efficiency and poor binding performance, and imprinted molecules are not easy to elute [Kryscio D R, Peppas N A. Critical review and perspective of macromolecularly imprinted polymers, Acta Biomaterialia, 2012, 8: 461–473]; (2) The choice of solvent is limited

In an organic solvent, although the hydrogen bond interaction between the template protein molecule and the monomer (so far, most protein molecular imprinting mainly relies on the hydrogen bond interaction between the template molecule and the functional monomer) is not disturbed, which is beneficial to the imprinting process , but most proteins have poor stability and solubility in organic solvents [Bossi A, Bonini F, Turner A P F, Piletsky S A. Molecularly imprinted polymers for the recognition of proteins: The state of the art, Biosensors and Bioelectronics, 2007, 22: 1131–1137]; if water is used as a solvent, although the protein is soluble in water, the hydrogen bond interaction between the template protein and the monomer will be competed and destroyed by water molecules, which is not conducive to the imprinting process; (3 ) Single monomer, small range of choices

As mentioned above, hydrogen bond interaction is still the main force basis of Western blotting, so the monomers used are mostly limited to acrylamide and acrylic monomers, and the blotting effect is affected.