Monolithic Chromatographic Materials:Preparation And Its Properties For Protein Separation

It is vitally important to develop the high-efficiency technique for separation of intact proteins.In recent years,capillary liquid chromatography has been paid much attention.However,the separation efficiency and selectivity of the traditional stationary phase is still limited.The present work focus on the development of new monolithic based stationary phase for capillary liquid chromatography so as to improve the separation performance of the intact proteins.The full text is divided into three chapters.The first chapter: The main technology for protein separation and the main chromatographic separation technology.The second chapter: In this chapter,ionic liquid based highly cross-linked organic polymer monolithic column poly(DPEPA-AVI)was prepared by "one pot" approach.The poly(DPEPA-AVI)column was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy,respectively.It exhibited typical reversed-phase liquid chromatography retention mechanism.Alkylbenzenes,polycyclic aromatic hydrocarbons,aromatic amines and standard protein samples were used to evaluate the chromatographic performance of the poly(DPEPA-AVI)column.The results shown that the poly(DPEPA-AVI)monolithic column has good potential for HPLC separation and we expect it can be further used for protein separation.The third chapter: In this chapter,DPEPA-POSS hybrid monolithic column was prepared for the first time by using "one-pot" approach.The DPEPA-POSS hybrid monolithic column was respectively characterized by Fourier transform infrared spectrometry,thermogravimetric analysis,scanning electron microscopy and nitrogen adsorption/desorption measurement.The new DPEPA-POSS hybrid monolith exhibited typical reversed-phase chromatography(RPLC)retention mechanism.The separation results show that the DPEPA-POSS hybrid capillary monolith has excellent column efficiency and excellent separation selectivity,when it was used for the separation of amides,thioureas and positional isomers of phenols.Subsequently,the intact proteins,egg white proteins and expressed BARD1 BRCT domains protein sample were isolated and compared to the commercialized C8 column and the separation efficiency was better than the commercial C8 with excellent selectivity.In the future work,the DPEPA-POSS hybrid monolithic column will be expected to play an important role in the separation and analysis of intact proteins.