A quantitative and selective chromatography method for determining coverages of multiple proteins on surfaces

Ombelli, Michela; Composto, Russell J.; Meng, Qing Cheng; Eckmann, David M.

doi:10.1016/j.jchromb.2005.09.002

Cited by 14 publications

(26 citation statements)

References 37 publications

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“…The large reductions in protein adsorption observed with the coated materials is in agreement with Ombelli et al's results [9] using single protein or a mixture of two proteins. This suggests that protein adsorption experiments of single or binary components are reliable data to predict phenomena in multi-component systems.…”

Section: Resultssupporting

confidence: 91%

“…This resistance continues up to two hours of plasma incubation, as depicted in Figures 6A and 6B. This is consistent with Ombelli et al's finding that dextranized silicon resists fibrinogen adsorption [9], as well as Holland's measurements using oligosaccharides [8]. Our results suggest that experiments with a single protein or with a two-protein mixture to study competition between select species yield important clues for understanding the more complex system of multi-component protein solutions such as human plasma.…”

Section: Resultssupporting

confidence: 90%

“…Dextran is chosen as a simple approach since sugars are prevalent constituents of model the endothelial glycocalyx layer. Previously, our laboratories have demonstrated dextranized silicon inhibits adsorption of bovine serum albumin, bovine fibrinogen and their binary mixture [9]. Marchant's group has covalently linked poly (vinyl amine) with dextran and show such surfactant polymer coating reduces platelet adhesion on polycarbonate surface [10].…”

Section: Introductionmentioning

confidence: 99%

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Human plasma protein adsorption onto dextranized surfaces: A two-dimensional electrophoresis and mass spectrometry study

Tsai

Tomczyk

Eckmann

et al. 2011

Colloids and Surfaces B: Biointerfaces

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Protein adsorption is fundamental to thrombosis and to the design of biocompatible materials. We report a two-dimensional electrophoresis and mass spectrometry study to characterize multiple human plasma proteins adsorbed onto four different types of model surfaces: silicon oxide, dextranized silicon, polyurethane and dextranized polyurethane. Dextran was grafted onto the surfaces of silicon and polyurethane to mimic the blood-contacting endothelial cell glycocalyx surface. Surface topography and hydrophobicity/hydrophilicity were determined and analyzed using atomic force microscopy and water contact angle measurements, respectively. Using twodimensional electrophoresis, we show that, relative to the unmodified surfaces, dextranization significantly inhibits the adsorption of several human plasma proteins including IGHG1 protein, fibrinogen, haptoglobin, Apo A-IV, Apo A-I, immunoglobulin, serum retinal-binding protein and truncated serum albumin. We further demonstrate the selectivity of plasma protein adsorbed onto the different functionalized surfaces and the potential to control and manipulate proteins adsorption on the surfaces of medical devices, implants and microfluidic devices. This result shows that adsorption experiments using a single protein or a binary mixture of proteins are consistent with competitive protein adsorption studies. In summary, these studies indicate that coating blood-contacting biomedical applications with dextran is an effective route to reduce thrombo-inflammatory responses and to surface-direct biological activities.

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Section: Resultssupporting

confidence: 91%

Section: Resultssupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

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Human plasma protein adsorption onto dextranized surfaces: A two-dimensional electrophoresis and mass spectrometry study

Tsai

Tomczyk

Eckmann

et al. 2011

Colloids and Surfaces B: Biointerfaces

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“…We have previously demonstrated that high-performance liquid chromatography (HPLC) is an effective tool for investigating protein adsorption from multicomponent mixtures onto biomaterial surfaces (Ombelli et al 2005). HPLC chromatograms yield both quantitative and qualitative information via peak area and retention time, respectively, for analysis of protein mixtures.…”

Section: Introductionmentioning

confidence: 99%

Competitive protein adsorption on polysaccharide and hyaluronate modified surfaces

et al. 2011

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We measured adsorption of bovine serum albumin (BSA) and fibrinogen (Fg) onto six distinct bare and dextran- and hyaluronate-modified silicon surfaces created using two dextran grafting densities and three hyaluronic acid (HA) sodium salts derived from human umbilical cord, rooster comb and streptococcus zooepidemicus. Film thickness and surface morphology depended on HA molecular weight and concentration. BSA coverage was enhanced on surfaces upon competitive adsorption of BSA:Fg mixtures. Dextranization differentially reduced protein adsorption onto surfaces based on oxidation state. Hyaluronization was demonstrated to provide the greatest resistance to protein coverage, equivalent to that of the most resistant dextranized surface. Resistance to protein adsorption was independent of the type of hyaluronic acid utilized. With changing bulk protein concentration from 20 to 40 µg ml−1 for each species, Fg coverage on silicon increased by 4×, whereas both BSA and Fg adsorption on dextran and HA were far less dependent of protein bulk concentration.

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“…Meanwhile, dextran, an a(1-6)-linked glycan, is nontoxic, water soluble and neutral, with multiple reactive sites available for subsequent functionalization for targeting ligands. Importantly, dextran has demonstrated outstanding properties against nonspecific protein adhesion by our group [25,26] and others [27][28][29]. Finally, dextran coated delivery systems have shown both reduced opsonisation by proteins in the medium [30] and reduced nonspecific hydrophobic interactions between the carrier and cell membranes.…”

Section: Discussionmentioning

confidence: 97%

Cellular Uptake and Intracellular Cargo Release From Dextran Based Nanogel Drug Carriers

Ferrer

Sobolewski²,

Composto

et al. 2013

Journal of Nanotechnology in Engineering and Medicine

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Nanogels (NG) hold great promise as a drug delivery platform. In this work, we examine the potential of lysozyme-dextran nanogels (LDNG) as drug carriers in vitro using two cell lines: a model target tissue, human umbilical cord vein endothelial cells (HUVEC) and a model of the mononuclear phagocyte system (phorbol 12-myristate 13-acetate (PMA)-stimulated THP-1 cells). The LDNG ($100 nm) were prepared with rhodaminelabel dextran (LRDNG) via Maillard reaction followed by heat-gelation reaction and were loaded with a fluorescent probe, 5-hexadecanoylaminofluorescein (HAF), as a mock drug. Epifluorescence microscopy confirmed rapid uptake of LRDNG by HUVEC. Although LysoTracker Green staining indicated a lysosomal fate for LRDNG, the mock drug cargo (HAF) diffused extensively inside the cell within 15 min. Flow cytometry and confocal microscopy indicated slow uptake of LRDNG in PMA-stimulated THP-1 cells, with only 41% of cells containing LRDNG after 24 h exposure. Finally, 24 h exposure to LRDNG did not affect the viability of either cell type at the dose studied (20 lg/ml). At a higher dose (200 lg/ml), LRDNG resulted in a marked loss of viability of HUVEC and THP-1, measuring 30% and 38%, respectively. Collectively, our results demonstrate the great potential of LRDNG as a drug delivery platform, combining simple production, rapid uptake and cargo release in target cells with "stealth" properties and low cytotoxicity.

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A quantitative and selective chromatography method for determining coverages of multiple proteins on surfaces

Cited by 14 publications

References 37 publications

Human plasma protein adsorption onto dextranized surfaces: A two-dimensional electrophoresis and mass spectrometry study

Human plasma protein adsorption onto dextranized surfaces: A two-dimensional electrophoresis and mass spectrometry study

Competitive protein adsorption on polysaccharide and hyaluronate modified surfaces

Cellular Uptake and Intracellular Cargo Release From Dextran Based Nanogel Drug Carriers

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