Preparation of enzyme functionalized polyHIPE monoliths for protein digestion and peptides analysis

来源 :第二十届全国色谱学术报告会 | 被引量 : 0次 | 上传用户:ZuoLuo
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  The properties of polyHIPEs were characterized for the excellent flow and mass transport combined with high surface area at the interconnected network.1 The preparation of polyHIPE foam also offers flexibility to introduce functional comonomer2 conveniently grafting at polyHIPE surface.The promising approach to modify polyHIPE with specially functional group is particularly successful for recyclable catalysts,supports for synthesis and biodegradable scaffolds.In our study,a typical single water-in-oil high internal phase emulsion(HIPE)containing polyglutaraldehyde(PGA)was used as templates to prepare interconnecting porous monolith with high enzyme loading capacity.The ratio of STY-DVB,volume fraction of the disperesd phase,the concentration of PGA in dispersed phase were investigated to obtain sufficient three-demension crosslinking polyHIPEs which had good permeability and mechanical strength for highest trypsin functionalization rate and bioactivity.The functionalized monoliths could effectively digest bovine serum albumin(BSA)and cytochrome(Cyt-C)(1 ml 0.2 mg/ml)with sequence coverage of 59%and 78%and had excellent digestion performance in ten times reusing.More importantly,the reusable monoliths could be easily regenerated by 4 mol/L HCl for breaking the-C=Nbond between the trypsin and PGA based on the Schiff base reaction followed by uploading fresh enzyme.This strategy is stable,efficient and can be extended to a variety of bioanalysis systems for protein digestion as long as the polyHIPE can be surface-activated by complexation with other kind of enzyme.
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