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[Langmuir] Spatially Addressable Chemoselective C-Terminal Ligation of an Intein Fusion Protein from a Complex Mixture to a Hydrazine-Terminated Surface
作者:Peng Yang, Stella M. Marinakos, and Ashutosh Chilkoti
关键字:doi. 10.1021/la104186n
论文来源:期刊
具体来源:Langmuir (Supramolecular Chemistry at Interfaces Special Issue)
发表时间:2010年

Protein immobilization on surfaces is useful in many areas of research, including biological characterization, antibody purification, and clinical diagnostics. A critical limitation in the development of protein microarrays and heterogeneous protein-based assays is the enormous amount of work and associated costs in the purification of proteins prior to their immobilization onto a surface. Methods to address this problem would simplify the development of interfacial diagnostics that use a protein as the recognition element. Herein, we describe an approach to the facile, site- specific immobilization of proteins on a surface without any preprocessing or sample purification steps that ligates an intein fusion protein at its C-terminus by reaction with a hydrazine group presented by a surface. Furthermore, we demonstrate that this methodology can directly immobilize a protein directly from cell lysate onto a protein-resistant surface. This methodology is also compatible with soft lithography and inkjet printing so that one or more proteins can be patterned on a surface without the need for purification.