Preparation Of Protein Molecularly Imprinted Polymers

In this work,a facile and efficient strategy was designed for preparation of molecularly imprinted polymers(c-MIPs)through acrylamied(AM)as monomer and N,N'-methylenebisacrylamide(MBA)as the cross-linker combined with the assembly(CD-PPRs)of cyclodextrin(CD)and polyvinyl alcohol(PVA)as assistant recognition.The MIPs was characterized by infrared spectroscopy(FT-IR),scanning electron microscope(SEM),thermogravimetric analysis(TG),ultraviolet spectrophotometer(UV)and sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)to study the selectivity,kinetics and reutilization of c-MIPs.Firstly,the supramolecular assembly(CD-PPRs)was assembled by CD as host molecules and PVA as guest molecules.The assembly and thermostability of CD-PPRs were tested by FT-IR and TG respectively.And UVwas used to explore the influence of CD-PPRs on template protein and the adsorption amount of template protein.It is found that the thermal stability of CD-PPRs was the highest in the proportion of 1:1.And acrylamied enter the protein gap and destroy the secondary structure of proteins,while the interaction of CD-PPRs with the protein surface stabilized the protein structure after introducing CD-PPRs.Secondly,bovine serum albumin(BSA),bovine hemoglobin(BHb),ovalbumin(OVA)and lysozyme(LYZ)was used as template molecules.The protein molecularly imprinted polymer(c-MIPs)was synthesized by free radical polymerization introducing the CD-PPRs.The results showed that CD-PPRs increased the molecular force of c-MIPs while protecting the structure of protein,and regular imprinted cavities of c-MIPs can be seen clearly different from other molecularly imprinted polymers on the surface of c-MIPs.In adsorption test,the template BSA was selected as the main research object.The adsorption performance test showed that the the maximum adsorption capacity of c-MIPs is 3.455 mg g-1 for BSA,and the adsorption of BSA reached a balance at 6 h.The adsorption process can be better described by the pseudo-second-order model,that is,the adsorption process of c-MIPs was controlled by chemical action and not limited by mass transfer resistance.What's more,the resultant materials exhibit good selectivity with a high imprinting factor(IF is 11.1)which was suitable for different template protein.And after repeated adsorption and desorption process,the utilization rate of c-MIPs is still 92%.Finally,the effect of cross-linking degree and CD-PPRs on the adsorption properties of the MIPs.When the ratio of monomer and crosslinker is 1:0.5,the maximum adsorption capacity of a-MIPs is 1.16 mg·g-1.And for c-MIPs,when the crosslinker increases to 1:0.7,the maximum adsorption capacity is 3.17 mg g-1.What's more,the adsorption capacity increased along with the concentration of CD-PPRs and decreased with the further increases of CD-PPRs.The polymers have maximum adsorption capacity when the the ratio of acrylamied and CD-PPRs is 1:0.5.And the adsorption capacity of c-MIPs was significantly higher than that of MIPs without CD-PPRs and blank molecularly imprinted polymer.It is proved that CD-PPRs can maintain the cavity shape and reduce the mass transfer resistance effect.