Title
Immobilization of proteins on carboxylic acid functionalized nanopatterns
Document Type
Article
Publication Date
2-1-2013
Abstract
The immobilization of proteins on nanopatterned surfaces was investigated using in situ atomic force microscopy (AFM) and ex situ infrared reflectance-absorption spectroscopy (IRAS). The AFM-based lithography technique of nanografting provided control of the size, geometry, and spatial placement of nanopatterns within self-assembled monolayers (SAMs). Square nanopatterns of carboxylate-terminated SAMs were inscribed within methyl-terminated octadecanethiolate SAMs and activated using carbodiimide/succinimide coupling chemistry. Staphylococcal protein A was immobilized on the activated nanopatterns before exposure to rabbit immunoglobulin G. In situ AFM was used to monitor changes in the topography and friction of the nanopatterns in solution upon protein immobilization. Complementary studies with ex situ IRAS confirmed the surface chemistry that occurred during the steps of SAM activation and subsequent protein immobilization on unpatterned samples. Since carbodiimide/succinimide coupling chemistry can be used for surface attachment of different biomolecules, this protocol shows promise for development of other aqueous-based studies for nanopatterned protein immobilization. © 2012 Springer-Verlag Berlin Heidelberg (outside the USA).
Publication Source (Journal or Book title)
Analytical and Bioanalytical Chemistry
First Page
1985
Last Page
1993
Recommended Citation
Ngunjiri, J., Stark, D., Tian, T., Briggman, K., & Garno, J. (2013). Immobilization of proteins on carboxylic acid functionalized nanopatterns. Analytical and Bioanalytical Chemistry, 405 (6), 1985-1993. https://doi.org/10.1007/s00216-012-6621-3