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

Publication ,  Journal Article
Yang, P; Marinakos, SM; Chilkoti, A
Published in: Langmuir : the ACS journal of surfaces and colloids
February 2011

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.

Duke Scholars

Published In

Langmuir : the ACS journal of surfaces and colloids

DOI

EISSN

1520-5827

ISSN

0743-7463

Publication Date

February 2011

Volume

27

Issue

4

Start / End Page

1463 / 1471

Related Subject Headings

  • Recombinant Fusion Proteins
  • Inteins
  • Immobilized Proteins
  • Hydrazines
  • Chemical Physics
 

Citation

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ICMJE
MLA
NLM
Yang, P., Marinakos, S. M., & Chilkoti, A. (2011). Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface. Langmuir : The ACS Journal of Surfaces and Colloids, 27(4), 1463–1471. https://doi.org/10.1021/la104186n
Yang, Peng, Stella M. Marinakos, and Ashutosh Chilkoti. “Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface.Langmuir : The ACS Journal of Surfaces and Colloids 27, no. 4 (February 2011): 1463–71. https://doi.org/10.1021/la104186n.
Yang P, Marinakos SM, Chilkoti A. Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface. Langmuir : the ACS journal of surfaces and colloids. 2011 Feb;27(4):1463–71.
Yang, Peng, et al. “Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface.Langmuir : The ACS Journal of Surfaces and Colloids, vol. 27, no. 4, Feb. 2011, pp. 1463–71. Epmc, doi:10.1021/la104186n.
Yang P, Marinakos SM, Chilkoti A. Spatially addressable chemoselective C-terminal ligation of an intein fusion protein from a complex mixture to a hydrazine-terminated surface. Langmuir : the ACS journal of surfaces and colloids. 2011 Feb;27(4):1463–1471.
Journal cover image

Published In

Langmuir : the ACS journal of surfaces and colloids

DOI

EISSN

1520-5827

ISSN

0743-7463

Publication Date

February 2011

Volume

27

Issue

4

Start / End Page

1463 / 1471

Related Subject Headings

  • Recombinant Fusion Proteins
  • Inteins
  • Immobilized Proteins
  • Hydrazines
  • Chemical Physics