Effect of Chain Density on Inhibition of Protein Adsorption by Poly(ethylene glycol) Based Coatings

Malmsten, Martin; Emoto, Kazuo; Alstine, James M. Van

doi:10.1006/jcis.1998.5513

Cited by 311 publications

(298 citation statements)

References 68 publications

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“…This is not totally surprising since at the isoelectric point (pI = 5) a rather large amount of BSA has been found to adsorb on cellulose surfaces [31]. In addition, similar results for the adsorption of BSA on non-cellulosic surfaces have been observed as a result of reduced solvency at the pI [25,46]. However, it should be noted that alkaline buffer (10 mM NaCl, pH 10) rinsing did not remove the bound BSA as would have been expected from the electrostatic repulsion between negatively charged BSA and NFC surfaces.…”

Section: Amine Reactivity Of Edc/nhs Activated Nfc Filmsmentioning

confidence: 52%

Surface Functionalized Nanofibrillar Cellulose (NFC) Film as a Platform for Immunoassays and Diagnostics

et al. 2012

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We introduce a new method to modify films of nanofibrillated cellulose (NFC) to produce non-porous, water-resistant substrates for diagnostics. First, water resistant NFC films were prepared from mechanically disintegrated NFC hydrogel, and then their surfaces were carboxylated via TEMPO-mediated oxidation. Next, the topologically functionalized film was activated via EDS/NHS chemistry, and its reactivity verified with bovine serum albumin and antihuman IgG. The surface carboxylation, EDC/NHS activation and the protein attachment were confirmed using quartz crystal microbalance with dissipation, contact angle measurements, conductometric titrations, X-ray photoelectron spectroscopy and fluorescence microscopy. The surface morphology of the prepared films was investigated using confocal laser scanning microscopy and atomic force microscopy. Finally, we demonstrate that antihuman IgG can be immobilized on the activated NFC surface using commercial piezoelectric inkjet printing.

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Section: Amine Reactivity Of Edc/nhs Activated Nfc Filmsmentioning

confidence: 52%

Surface Functionalized Nanofibrillar Cellulose (NFC) Film as a Platform for Immunoassays and Diagnostics

et al. 2012

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“…The concept of PEGylated proteins consisting of conjugated but distinct spheroids is not in keeping with biomedical results (32,36,37) which suggest that PEGylation even with relatively small MW PEG's can dramatically alter a protein's average surface properties. Such results are more in keeping with the single spheroid concept assumed in the present study in deriving equation (11).…”

Section: Modelmentioning

confidence: 99%

Prediction of the Viscosity Radius and the Size Exclusion Chromatography Behavior of PEGylated Proteins

2004

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Size exclusion chromatography (SEC) was used to determine the viscosity radii of equivalent spheres for proteins covalently grafted with poly(ethylene glycol) (PEG). The viscosity radius of such PEGylated proteins was found to depend on the molecular weight of the native protein and the total weight of grafted PEG but not on PEG molecular weight, or PEG-toprotein molar grafting ratio. Results suggest grafted PEG's form a dynamic layer over the surface of proteins. The geometry of this layer results in a surface area to volume ratio approximately equal to that of a randomly coiled PEG molecule of equivalent total molecular weight. Two simple methods are given to predict the viscosity radius of PEGylated proteins.Both methods accurately predicted (3% absolute error) the viscosity radii of various PEGproteins produced using three native proteins, α-lactalbumin (14.2 kDa MW), β−lactoglobulin dimer (37.4 kDa MW) and bovine serum albumin (66.7 kDa MW), three PEG reagents (2400, 5600, and 22500 MW), and molar grafting ratios of 0 to 8. Accurate viscosity radius prediction allows calculation of the distribution coefficient, K av , for PEGproteins in SEC. The suitability of a given SEC step for the analytical or preparative fractionation of different PEGylated protein mixtures may therefore be assessed mathematically. The methods and results offer insight to several factors related to the production, purification, and uses of PEGylated proteins.

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“…25,26 As demonstrated theoretically [26][27][28][29] and experimentally [30][31][32][33][34][35] protein resistance of PEG-based coatings depend on both chain length and density, the product of which determines the thickness of the PEG adlayer. Protein resistance becomes increasingly efficient the more the PEG chains overlap, 33 and longer chains have been found to have the ability to fill in the gaps between less densely grafted PEG chains. 34 Theoretical studies have revealed two modes of protein adsorption: 29 ͑i͒ primary adsorption at the PEG-substrate interface and ͑ii͒ secondary adsorption at the top of the PEG brush.…”

Section: Introductionmentioning

confidence: 99%

Phagocytosis of poly(L-lysine)-graft-poly (ethylene glycol) coated microspheres by antigen presenting cells: Impact of grafting ratio and poly (ethylene glycol) chain length on cellular recognition

et al. 2006

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Microparticulate carrier systems have significant potential for antigen delivery. The authors studied how microspheres coated with the polycationic copolymer poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) can be protected against unspecific phagocytosis by antigen presenting cells, a prerequisite for selective targeting of phagocytic receptors. For this aim the authors explored the influence of PLL-g-PEG architecture on recognition of coated microspheres by antigen presenting cells with regard to both grafting ratio and molecular weight of the grafted PEG chains. Carboxylated polystyrene microspheres (5 μm) were coated with a small library of PLL-g-PEG polymers with PLL backbones of 20 kDa, grafting ratios from 2 to 20, and PEG side chains of 1–5 kDa. The coated microspheres were characterized by their ζ-potential and resistance to IgG adsorption. Phagocytosis of these microspheres by human monocyte derived dendritic cells (DCs) and macrophages (MΦ) was quantified by phase contrast microscopy and by analysis of the cells’ side scattering in a flow cytometer. Generally, increasing grafting ratios impaired the protein resistance of coated microspheres, leading to higher phagocytosis rates. For DC, long PEG chains of 5 kDa decreased the phagocytosis of coated microspheres even in the case of considerable IgG adsorption. In addition, preferential adsorption of dysopsonins is discussed as another factor for decreased phagocytosis rates. For comparison, the authors studied the cellular adhesion of DC and Mζ to PLL-g-PEG coated microscopy slides. Remarkably, DC and Mζ were found to adhere to relatively protein-resistant PLL-g-PEG adlayers, whereas phagocytosis of microspheres coated with the same copolymers was inefficient. Overall, PLL(20)-[3.5]-PEG(2) was identified as the optimal copolymer to ensure resistance to both phagocytosis and cell adhesion. Finally, the authors studied coatings made from binary mixtures of PLL-g-PEG type copolymers that led to microspheres with combined properties. This enables future studies on cell targeting with ligand modified copolymers.

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Effect of Chain Density on Inhibition of Protein Adsorption by Poly(ethylene glycol) Based Coatings

Cited by 311 publications

References 68 publications

Surface Functionalized Nanofibrillar Cellulose (NFC) Film as a Platform for Immunoassays and Diagnostics

Surface Functionalized Nanofibrillar Cellulose (NFC) Film as a Platform for Immunoassays and Diagnostics

Prediction of the Viscosity Radius and the Size Exclusion Chromatography Behavior of PEGylated Proteins

Phagocytosis of poly(L-lysine)-graft-poly (ethylene glycol) coated microspheres by antigen presenting cells: Impact of grafting ratio and poly (ethylene glycol) chain length on cellular recognition

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