1. High performance aptamer affinity chromatography for single-step selective extraction and screening of basic protein lysozyme
- Author
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Han, Bin, Zhao, Chao, Yin, Junfa, and Wang, Hailin
- Subjects
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AFFINITY chromatography , *DNA , *APTAMERS , *GLYCIDYL methacrylate , *ADDITION polymerization , *LYSOZYMES , *POLYMETHACRYLATES , *AMINO acid sequence - Abstract
Abstract: A DNA aptamer based high-performance affinity chromatography is developed for selective extraction and screening of a basic protein lysozyme. First, a poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolithic column was synthesized in situ by thermally initiated radical polymerization, and then an anti-lysozyme DNA aptamer was covalently immobilized on the surface of the monolith through a 16-atom spacer arm. The target protein lysozyme but non-target proteins can be trapped by the immobilized anti-lysozyme DNA aptamer. In contrast, lysozyme cannot be trapped by the immobilized oligodeoxynucleotide that does not contain the sequence of the anti-lysozyme DNA aptamer. The study clearly demonstrates the trapping of lysozyme by the immobilized anti-lysozyme DNA aptamer is mainly due to specific recognition rather than simple electrostatic interaction of positively charged protein and the negatively charged DNA. The inter-day precision was determined as 0.8% for migration time and 4.2% for peak area, respectively. By the use of aptamer affinity monolith, a screening strategy is developed to selectively extract lysozyme from chicken egg white, showing the advantages of high efficiency, low cost and ease-of-operation. [Copyright &y& Elsevier]
- Published
- 2012
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