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Immobilized Streptavidin Gradients as Bioconjugation Platforms

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Bryan R. Coad*†‡, Krasimir Vasilev‡, Kerrilyn R. Diener§

, John D. Hayball§

#, Robert D. Short‡, and Hans J. Griesser†

^† Ian Wark Research Institute, University of South Australia, Mawson Lakes, 5095 SA, Australia

^‡ Mawson Institute, University of South Australia, Mawson Lakes, 5095 SA, Australia

^§ Experimental Therapeutics Laboratory, Hanson Institute, Royal Adelaide Hospital, Adelaide SA 5000, Australia

School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide SA 5005, Australia

Sansom Institute, University of South Australia, Adelaide SA 5000, Australia

^# School of Medicine, University of Adelaide, Adelaide SA 5000, Australia

Langmuir, 2012, 28 (5), pp 2710–2717

DOI: 10.1021/la204714p

Publication Date (Web): January 11, 2012

*E-mail: Bryan.Coad@unisa.edu.au.

Section:

Biochemical Methods

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.

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