Affinity selection of target cells from cell surface displayed libraries: a novel procedure using thermo-responsive magnetic nanoparticles.

作者: H. Furukawa , R. Shimojyo , N. Ohnishi , H. Fukuda , A. Kondo

DOI: 10.1007/S00253-003-1330-7

关键词:

摘要: Biotinylated thermo-responsive magnetic nanoparticles for use in affinity selection from yeast cell surface display libraries were prepared by coating magnetite with a polymer consisting of N-isopropyl acrylamide and biotin derivative. These particles showed reversible transition between flocculation dispersion at around the lower critical solution temperature 30°C, above which flocculated particles—which absorbed large amount avidin due to their area—were quickly separable magnet. The model library was constructed mixing control cells target co-displaying IgG binding protein (ZZ) enhanced green fluorescence protein. subsequently added library, efficiently enriched biotinylated avidin-biotin sandwich ZZ-IgG interaction. few (0.001%) up 100% only 5 days an procedure repeated four times. This novel method based on system could fulfill wide range applications analysis protein-protein interactions rapid isolation biomolecules.

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