Preparation of a Tetrazolyl Monolithic Column via the Combination of ATRP and Click Chemistry for the Separation of Proteins

Tetrazolyl monolithic column is first prepared through the combination of atom transfer radical polymerization (ATRP) and "click chemistry" technique. In the ATRP fabrication process, vinyl ester resin is used as both the monomer and the cross-linking agent, and cetyl alcohol is used as the porogen, carbon tetrachloride as the initiator and ferrous chloride as the catalyst. The monolith is modified by click chemistry, which forms the tetrazolyl monolithic column. The chemical group of the prepared monolith is assayed by infrared spectroscopy, and its internal morphology is investigated by scanning electron microscopy. The pore size distribution is determined by a mercury porosimeter. What is more, the monolithic column was used as the stationary phase during high-performance liquid chromatography. Moreover, the monolith is used to separate lysozyme (Lys) from egg whites in a short time period (4 min). At the same time, the influences of buffer concentration and pH value on the elution of Lys are investigated. In addition, human serum albumin is successfully separated from human plasma by the prepared monoliths. The results show that click chemistry is an efficient method to modify the monoliths prepared by ATRP.