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A thermosensitive monolithic column as an artificial antibody for the on-line selective separation of the protein
Authors:Qin Lei  He Xi-Wen  Jia Man  Li Wen-You  Zhang Yu-Kui
Institution:Department of Chemistry, Nankai University, Tianjin 300071, China.
Abstract:The main objective of this study was to develop a new methodology for the preparation of a protein (antigen) that is a molecularly imprinted polymer (MIP, an artificial antibody) modified onto the surface of a silica skeleton in which the resulting stationary phase is thermosensitive. The silica monolithic skeleton with vinyl groups was synthesized in a stainless-steel column by using a mild one-step sol-gel process with two types of precursor: methyltrimethoxysilane (MTMS) and γ-methacryloxypropyltrimethoxysilane (γ-MAPS). Subsequently, three types of the thermosensitive protein MIP were anchored onto the surface of the silica skeleton to prepare the MIP monoliths, which were systematically investigated for back pressure and separation ability at different temperatures to establish good imprinting conditions. Under the optimized imprinting conditions, the chromatographic behavior of the thermosensitive MIP monolith exhibited strong retention ability for the lysozyme template (target antigen) in relation to the nonimprinting monolith (NIP monolith). The imprinting factor (IF) for lysozyme reached 3.48 at 20 °C. Moreover, this new type of artificial antibody displayed favorable binding characteristics for lysozyme over competitive proteins and was further evaluated to selectively separate lysozyme in a real sample by using an on-line method. The run-to-run and column-to-column repeatability measurements of the thermosensitive MIP monoliths were also satisfactory.
Keywords:antigens  imprinting  monolith columns  proteins  thermosensitivity
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