Issue 24, 2011

Localized entrapment of green fluorescent protein within nanostructured polymer films

Abstract

Protein entrapment within ultrathin polymer matrices is of significant interest for applications in biosensing, drug delivery, and bioconversion; however, it remains a big challenge due to insufficient control over protein distribution inside the matrices. We report on nanostructured protein-polyelectrolyte (PE) films obtained through localized incorporation of green fluorescent protein (GFP) within poly(styrene sulfonate)/poly(allylamine hydrochloride) matrices assembled via the spin-assisted layer-by-layer method. Film compositional and structural analyses were performed by using a combination of techniques such as neutron and X-ray reflectometry, circular dichroism (CD), atomic force microscopy (AFM), spectroscopic ellipsometry, and Attenuated Total Reflection Fourier transform infrared spectroscopy (ATR-FTIR). By using deuterated GFP as a marker for neutron scattering contrast we have inferred the architecture of the films both in vertical and lateral directions. We found that films assembled with a single GFP layer confined at various distances from the substrate exhibit a strong localization of the GFP layer without intermixing into the PE matrix. The GFP volume fraction approached the maximum value of a monolayer packing density of randomly oriented GFP molecules. However, partial intermixing of the GFP with polymeric material is evidenced in multiple-GFP layer films which showed alternating protein-rich and protein-deficient regions. Our results yield new insights into the organization of immobilized proteins within polyelectrolyte matrices and open opportunities for fabrication of protein-containing films with well-organized structure and controllable function, a crucial requirement for advanced sensing applications.

Graphical abstract: Localized entrapment of green fluorescent protein within nanostructured polymer films

Supplementary files

Article information

Article type
Paper
Submitted
09 Sep 2011
Accepted
05 Oct 2011
First published
21 Oct 2011

Soft Matter, 2011,7, 11453-11463

Localized entrapment of green fluorescent protein within nanostructured polymer films

V. Kozlovskaya, J. F. Ankner, H. O'Neill, Q. Zhang and E. Kharlampieva, Soft Matter, 2011, 7, 11453 DOI: 10.1039/C1SM06710A

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