Research Of Ion Exchange Chromatography Media For Protein Chromatography Supported By Polyacrylamide And Polyvinyl Alcohol

Ion-exchange chromatography continues to be dominant in the recovery, separation, and purification of proteins, particularly on the industrial process. As a result, there is considerable interest in improving the design and efficiency of ion-exchange adsorbents suitable for preparative and process chromatography. Polyacrylamide (PAM) has remarkable hydrophilicity, biocompatibility and chemical reactivity. Commercial PAM resin has already been used as a media of gel chromatography, which could resist the corrosion caused by microbes.Polyacrylamide resin with 20% cross-linking degree was prepared by inverse suspension polymerization. Aminomethylated polyacrylamide (APAM), a highly hydrophilic weak anion exchange resin, was prepared by modification of PAM with Mannich reaction. A highly hydrophilic strong cation exchange resin, sulfomethylated polyacrylamide (SPAM), was prepared by sulfomethylation of PAM. The influence of Time, pH and Temperature on the properties of the resins was determined.Polyvinyl acetate resin was prepared by suspension polymerization, and modified to carboxymethylated polyvinyl alcohol resin (PVAONCB) after several steps of reaction. The property of PVAONCB was characterized.The absorption behaviors of proteins on APAM, SPAM and PVAONCB have been primarily studied. The result indicate that the adsorption of protein on those resins lie on the electrostatic interaction between proteins and resins, and the adsorption binding capacity and binding strength is highly related to the surface charge density of the resin. The linear fitting curves show that BSA adsorption on those resins fitted Langmuir equation well.The separation efficiency of common proteins on APAM, SPAM and PVAONCB operated on manually assembled chromatography equipment were determined. The results show that: (1) APAM resin is suit for protein separation. The separation of BSA and Hemoglobin could be accomplished with salt concentration in elution increased gradually. (2) The interaction between protein and SPAM resin is too strong. So, the elution of proteins could not be accomplished by gradient elution with the concentration of salt increased gradually but with the pH of elution buffer increased orderly. (3)The retention time of common proteins on PVAONCB packing column is different, but the binding capacity and the resolving capacity are comparatively low, which may due to the shortage of charged functional group on the surface of resin.