Preparation method of sensitive western blotting material based on block macromolecular chain monomer

Abstract

The invention discloses a method for preparing a sensitive protein imprinting material based on a block macromolecular chain monomer, which specifically comprises the following steps: Step 1, O-ethyl-S-(1-phenylethyl) dithio Carbonate and sensitive monomers are dissolved in an organic solvent, and react under anaerobic conditions to obtain block macromolecular chain precursor 1; step 2, respectively add block macromolecular chain precursor 1 and hydroxyethyl acrylate to the organic solvent Esters, zwitterionic monomers and initiators are reacted under anaerobic conditions to obtain a block copolymerization macromolecular chain precursor 2; step 3, reacting the obtained block copolymerization macromolecular chain precursor 2 with acryloyl chloride to obtain Macromolecular functional monomer; step 4, assembling the protein and macromolecular functional monomer in phosphate buffer solution, adding ammonium persulfate and tetramethylethylenediamine to obtain a western blot hydrogel. Macromolecular chain monomers can obtain imprinted hole structures with highly specific interactions in molecularly imprinted materials.

Description

Preparation method of sensitive western blotting material based on block macromolecular chain monomer technical field The invention belongs to the field of polymer materials technology, and relates to a sensitive Western blotting material based on block macromolecular chain monomers. material preparation method. Background technique Proteomics is the study of the characteristics of proteins at different scales, including protein expression, protein translation Translation and protein-protein interactions have received extensive attention in recent years. by healthy individuals and pathological The proteomes of individuals were compared and several protein biomarkers associated with pathology were found. These proteins can be New molecular targets for drug design or molecular markers for early diagnosis of diseases. In order to achieve the separation or separation of target proteins Analysis, immunoassay based on antigen-antibody specific recognition is considere...

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Problems solved by technology

In addition, Krysico believes that the structural destruction of proteins during imprinting is mainly caused by the destruction of functional monomers and cross-linking agents (Kryscio D R, Fleming M Q, Peppas N A. Protein conformational studies for macromolecularly imprinted polymers[J].Macromolecular Bioscience, 2012, 12(8):1137-1144), the smaller molecular volume of traditional functional monomers and cross-linking agents relative to proteins endows them with strong flexibility, making it easier to penetrate into proteins, thus Destroys the hydrogen bonds that maintain the stability of the structure and conformation inside the protein, resulting in changes in the protein structure

On the other hand, the protein structure is complex, and the surface is rich in amino acid residues of various properties, so it is inevitable that it can interact with the surface of the material other than the imprinted hole, resulting in non-specific adsorption and a decrease in the recognition of the imprinted material.

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