title: High performance aptamer affinity chromatography for single-step selective extraction and screening of basic protein lysozyme.
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.
authors: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100086, PR China.
authors: Bin
authors: B
authors: Han
authors: Chao
authors: C
authors: Zhao
authors: Junfa
authors: J
authors: Yin
authors: Hailin
authors: H
authors: Wang
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008768:Methylmethacrylates; D009113:Muramidase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D016062:Porosity; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D052157:Aptamers, Nucleotide; D008768:Methylmethacrylates; C020347:GMA-EDMA; D009113:Muramidase