title: Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface.
abstract: 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 for 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.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina 27705, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Yang
authors.forename: Stella M
authors.initials: SM
authors.lastname: Marinakos
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Chilkoti
mesh_terms: D006834:Hydrazines / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D047668:Inteins; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
chemical_list: D006834:Hydrazines; D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; C029424:hydrazine