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

Abstract

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.

Competitive protein adsorption on polysaccharide and hyaluronate modified surfaces

“…Since the dextran layer thickness can vary, detection of nitrogen from within the APTES layer is possible. The amine groups may be exposed within the dextran layer, since the AFM images as seen, for instance, in (Tsai et al 2011a) for surfaces prepared identically can appear patchy. For our hyaluronized silicon surfaces made at the NaHA concentration of 1.2 mg ml −1 , XPS used under the same conditions as for dextran showed the ratio of C to N on average to be 10.8 (streptococcus zooepidemicus NaHA), 11.2 (human NaHA), and 11.4 (rooster comb NaHA).…”

Hyaluronan and dextran modified tubes resist cellular activation with blood contact

“…Oxidized dextran macromolecules were grafted onto the aminated tubes using 0.015 gm of NaBH 3 CN for 1 day. Dextrans were oxidized using the same procedure as described in our previous publications for silicon wafers [7;24;25;28–32;41;42]. The dextranized surface for this experiment was obtained by oxidizing dextran for 0.5 hr, with additional methodological detail having been given previously [25;30].…”

“…Results of surface characterization including these types of measures as well as zeta potential analysis of planar dextranized and hyaluronized surfaces have previously been reported [25;30;41;42]. …”

Quantitation of nonspecific protein adsorption at solid–liquid interfaces for single-cell proteomics

“…Vogler et al have summarized such applications 117,118 . When using 2-dimensional (2D)-gel coupled with MS, the technique can tackle proteome-wide protein adsorption 119 . The drawback of the method is its ex situ analysis that has limited its application from conducting fast kinetic studies.…”