Immobilized Streptavidin Gradients as Bioconjugation Platforms

Bryan R. Coad, Krasimir Vasilev, Kerrilyn R. Diener, John D. Hayball, Robert D. Short, Hans J. Griesser

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

Surface density gradients of streptavidin (SAV) were created on solid surfaces and demonstrated functionality as a bioconjugation platform. The surface density of immobilized streptavidin steadily increased in one dimension from 0 to 235 ng cm -2 over a distance of 10 mm. The density of coupled protein was controlled by its immobilization onto a polymer surface bearing a gradient of aldehyde group density, onto which SAV was covalently linked using spontaneous imine bond formation between surface aldehyde functional groups and primary amine groups on the protein. As a control, human serum albumin was immobilized in the same manner. The gradient density of aldehyde groups was created using a method of simultaneous plasma copolymerization of ethanol and propionaldehyde. Control over the surface density of aldehyde groups was achieved by manipulating the flow rates of these vapors while moving a mask across substrates during plasma discharge. Immobilized SAV was able to bind biotinylated probes, indicating that the protein retained its functionality after being immobilized. This plasma polymerization technique conveniently allows virtually any substrate to be equipped with tunable protein gradients and provides a widely applicable method for bioconjugation to study effects arising from controllable surface densities of proteins.

Original languageEnglish
Pages (from-to)2710-2717
Number of pages8
JournalLangmuir
Volume28
Issue number5
DOIs
Publication statusPublished - 7 Feb 2012
Externally publishedYes

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