Published: March 07, 2011 r2011 American Chemical Society 1058 dx.doi.org/10.1021/bm101406m | Biomacromolecules 2011, 12, 1058–1066 ARTICLE pubs.acs.org/Biomac Substrate-Independent Approach for the Generation of Functional Protein Resistant Surfaces Cesar Rodriguez-Emmenegger,* ,†,‡ Ond  rej Kyli  an, § Milan Houska, † Eduard Brynda, † Anna Artemenko, § Jaroslav Kousal, § Aldo Bologna Alles, ‡ and Hynek Biederman § † Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, v.v.i., Czech Republic ‡ College of Engineering, Universidad de la Republica, Uruguay § Faculty of Mathematics and Physics, Charles University in Prague, Czech Republic b S Supporting Information ’ INTRODUCTION Protein fouling in complex biological fluids, particularly, blood, plasma, and serum, is an adverse event that can impair the properties or functions of various biotechnological and biomedical devices. 13 Some examples include stopping flow through separation columns and porous membranes, 4 nonspe- cific response of affinity biosensors, 2,3,5 reduced circulation time of nanocarriers in bloodstream due to colloidal instability or opsonization, 610 bacteria attachment on contact lenses 11 and synthetic grafts, 12 or disabling of cardiovascular devices by thrombus formation. 12,13 Thus, the development of a technology by which antifouling interfaces between biological media and various types of materials can be prepared is an important challenge for contemporary research. Current surface modifications with antifouling self-assembled monolayers (SAMs), 14 grafted polymer layers, and polymer brushes reduce considerably or suppress the adsorption from single protein solutions. The authors have often claimed that they have obtained perfectly antifouling, 15 superlow fouling, 16 ultralow fouling, 17 and even nonfouling 18 surfaces because they had not observed any adsorption from solutions of the main plasma proteins, human serum albumin (HSA), and fibrinogen (Fbg). However, a reduction or even total prevention of the adsorption of the main blood plasma proteins (HSA and Fbg), immunoglo- bulin G (IgG) or lysozyme (Lys), is not evidence that the surface is resistant to blood plasma. 3,19,20 Only much fewer works showed a reduction of the fouling from blood plasma. 13 Grafted carboxymethyldextrane or polyethyleneglycol have been used, but just a minor decrease in blood plasma fouling was reached. Theoretical treatment predicted enhanced antifouling properties Received: November 24, 2010 Revised: March 2, 2011 ABSTRACT: A new route for coating various substrates with antifouling polymer layers was developed. It consisted in deposition of an amino-rich adhesion layer by means of RF magnetron sputtering of Nylon 6,6 followed by the well- controlled, surface-initiated atom transfer radical polymeriza- tion of antifouling polymer brushes initiated by bromoisobuty- rate covalently attached to amino groups present in the adhesion layer. Polymer brushes of hydroxy- and methoxy- capped oligoethyleneglycol methacrylate and carboxybetaine acrylamide were grafted from bromoisobutyrate initiator at- tached to a 15 nm thick amino-rich adhesion layer deposited on gold, silicon, polypropylene, and titaniumaluminum vanadium alloy surfaces. Well-controlled polymerization ki- netics made it possible to control the thickness of the brushes at a nanometer scale. Zero fouling from single protein solutions and a reduction of more than 90% in the fouling from blood plasma observed on the uncoated surfaces was achieved. The feasibility of functionalization with bioactive compounds was tested by covalent attachment of streptavidin onto poly(oligoethylene glycol methacrylate) brush and subsequent immobilization of model antibodies and oligonucleotides. The procedure is nondestructive and does not require any chemical preactivation or the presence of reactive groups on the substrate surface. Contrary to current antifouling modifications, the developed coating can be built on various classes of substrates and preserves its antifouling properties even in undiluted blood plasma. The new technique might be used for fabrication of biotechnological and biomedical devices with tailor-made functions that will not be impaired by fouling from ambient biological media.