Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Yang, P; Marinakos, SM; Chilkoti, A

Published Date

  • February 2011

Published In

Volume / Issue

  • 27 / 4

Start / End Page

  • 1463 - 1471

PubMed ID

  • 21142101

Pubmed Central ID

  • PMC3189817

Electronic International Standard Serial Number (EISSN)

  • 1520-5827

International Standard Serial Number (ISSN)

  • 0743-7463

Digital Object Identifier (DOI)

  • 10.1021/la104186n

Language

  • eng