期刊
CHEMISTRY-A EUROPEAN JOURNAL
卷 17, 期 5, 页码 1696-1704出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/chem.201000875
关键词
antigens; imprinting; monolith columns; proteins; thermosensitivity
资金
- National Basic Research Program of China (973 Program) [2007CB914100, 2006CB705703]
- National Nature Science Foundation of China [20875049, 21075069]
- Tianjin Natural Science Foundation [06YFJMJC02800]
- Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences
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 gamma-methacryloxypropyltrimethoxysilane (gamma-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 non-imprinting 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.
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