Immobilization and surface characterization of NeutrAvidin biotin-binding protein on different hydrogel interlayers

For a number of potential applications, it is desirable to immobilize avidin class molecules onto solid supports and exploit their ability to bind biotinylated molecules with high affinity. NeutrAvidin molecules were surface immobilized in various ways. In this study, NeutrAvidin was covalently attached by carbodiimide chemistry onto carboxyl groups of polyacrylic acid and carboxymethyl-dextran hydrogel interlayers. A third strategy involved the affinity "docking" of NeutrAvidin onto a biotinylated poly(ethylene glycol) interlayer. These three interlayers were selected for their low nonspecific binding of proteins, which was expected to minimize surface binding of NeutrAvidin by nonspecific interfacial adsorption. X-ray photoelectron spectroscopy (XPS) analyses allowed detailed characterization of the multilayer fabrication steps. An ELISA assay was used to measure NeutrAvidin activity, which varied with the surface immobilization route. Atomic force microcopy (AFM) force measurements showed that the hydrogel interlayer contributed to a repulsive force and verified the specific interaction between biotinylated AFM tips and the NeutrAvidin surfaces. When a solution of free biotin was injected into the AFM liquid cell, the force curve changed substantially and became identical to that recorded between surfaces carrying no NeutrAvidin, indicating that the free solution biotin had displaced NeutrAvidin proteins off the PEG-biotin layer.     

Characterization of low-fouling ethylene glycol containing plasma polymer films

Low-protein-fouling poly(ethylene glycol) (PEG-like) plasma polymer films were prepared using radio frequency glow discharge polymerization of diethylene glycol dimethyl ether (DGpp) on top of a heptylamine plasma polymer primer layer. By varying the plasma deposition conditions, the chemistry of the DGpp film was influenced, especially in regard to the level of ether content, which in turn influenced the relative levels of bovine serum albumin and lysozyme protein fouling. Surface potential measurements indicated that these surfaces carried a net negative charge. While protein fouling remained low ( approximately 10 ng/cm2), there was a slightly higher level of the positively charged protein adsorbed on these films than the negative protein. The interaction forces measured between a silica spherical surface on both "high"- and "low"-protein-fouling DGpp films were all repulsive and short ranged (2-3 nm). There was no correlation between the surface forces measured for high- and low-protein-fouling DGpp films. Thus, it appears that enthalpic effects are very important in reducing protein adsorption. We therefore conclude that it is the concentration of residual, ethylene glycol containing species that are the crucial parameter determining protein resistance due to a combination of both entropic and enthalpic effects.     

On the structure of small strength‐2 covering arrays

A covering array $\text{CA}\left(N;t,k,v\right)$ of strength $t$ is an $N\times k$ array of symbols from an alphabet of size $v$ such that in every $N\times t$ subarray, every $t$‐tuple occurs in at least one row. A covering array is optimal if it has the smallest possible $N$ for given $t$, $k$, and $v$, and uniform if every symbol occurs $?N∕v?$ or $?N∕v?$ times in every column. Before this paper, the only known optimal covering arrays for $t=2$ were orthogonal arrays, covering arrays with $v=2$ constructed from Sperner's Theorem and the Erd?s‐Ko‐Rado Theorem, and 11 other parameter sets with $v>2$ and $N>v^2$. In all these cases, there is a uniform covering array with the optimal size. It has been conjectured that there exists a uniform covering array of optimal size for all parameters. In this paper, a new lower bound as well as structural constraints for small uniform strength‐2 covering arrays is given. Moreover, covering arrays with small parameters are studied computationally. The size of an optimal strength‐2 covering array with $v>2$ and $N>v^2$ is now known for 21 parameter sets. Our constructive results continue to support the conjecture.     

One‐Reactant Photografting of ATRP Initiators for Surface‐Initiated Polymerization

Self‐initiated photografting polymerization is used to couple the polymerizable initiator monomer 2‐(2‐chloropropanoyloxy)ethyl acrylate to a range of polymeric substrates. The technique requires only UV light to couple the initiator to surfaces. The initiator surface density can be varied by inclusion of a diluent monomer or via selection of initiator and irradiation parameters. The functionality of the initiator surface is demonstrated by subsequent surface‐initiated atom transfer radical polymerization. Surfaces are characterized by x‐ray photoelectron spectroscopy (XPS), ellipsometry, and atomic force microscopy (AFM), and UV‐induced changes to the initiator are assessed by ¹H NMR and gel permeation chromatography (GPC). This is the first time this one‐reactant one‐step technique has been demonstrated for creating an initiator surface of variable density.