Specific Recognition Of Protein Imprinted Coomassie Brilliant Blue Chromogenic Hydrogel To Target Molecule

For a long time,proteins are the core substances in the life activities of organisms,and they play an important role in catalysis,transportation and immune regulation.Therefore,the separation and detection of proteins have attracted more attention.Due to the limitations of the traditional single protein detection methods,such as poor specificity and cumbersome operation,more and more scientists are engaged in the field of protein detection.Lowry method has attracted much attention because of its rapid detection and other advantages,but the lack of specificity between dyes and proteins is a deficiency of this method.On the other side,commonly used immunoblotting test has good specificity in protein detection but is suffering from the difficulties in antibodies preparation and preservation,which leads to high cost and difficulty in promotion.Molecular imprinting technology derived from immunization can prepare antibody analogues(molecularly imprinted polymers).The use of this technology has the advantages of simple operation and materials easy storage.The emergence of this technology can well solve the shortcomings of immunoblotting test.Therefore,the mechanism of coomassie brilliant blue and protein interaction was studied in this paper.Finally,it was used to guide the combination of Lowry protein assay method and molecular imprinting technology to prepare a molecularly imprinted chromogenic hydrogel.Firstly,we studied the effect of pH on coomassie brilliant blue solution color,absorption peak and the interaction between coomassie brilliant blue and protein.We investigated the difference of chromogenic laws caused by changing coomassie brilliant blue and BSA concentration.The results showed that there were three components in coomassie brilliant blue molecular and all of them can interacted with bovine serum albumin.The absorption peak of the association complex was around 610 n..The interaction between double protonated cation components and proteins can be affected due to low pH.The best chromogenic effect of coomassie brnlliant blue to protein was obtained at pH 0.6.Secondly,we investigated and analyzed the gel matrix,quantitative detection methods and gel synthesis process in this paper.The results indicated that the internal molecular structure of the polyacrylamide hydrogel did not affect the color development mechanism of coomassie brilliant blue due to good transparency of gel.The use of digital number of optical image to quantitative determination had the advantages of simple operation,simple equipment.The best synthesis process was 20 w/v%monomer concentration,2 wt%crosslinker to total monomers and 0.1 mol%initiator to total monomers.Finally,we synthesized the imprinted polymer and the non-imprinted polymer under the best synthesis process and investigated the elution effect of different eluents and the specific recognition performance of imprinted hydrogel to proteins.The results showed that 5 w/v%SDS-10 v/v%HAc solution was the best eluents and the imprinted polymer had a certain specific recognition ability for the target protein.