Identification of proteins adsorbed to hemodialyser membranes from heparinized plasma

Cornelius, Rena M.; Brash, John L.

doi:10.1163/156856293x00573

Cited by 68 publications

(30 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cornelius and Brash [43] demonstrated that C3 was present in SDS eludates after plasma incubations of a number of hemodialysis membranes. In accordance with this, we show in the present study that SDS has a dramatic washing effect on different complement activator surfaces.…”

Section: Discussionmentioning

confidence: 99%

On the binding of complement to solid artificial surfaces in vitro

et al. 2002

View full text Add to dashboard Cite

Since the realization of a complement activation capacity by artificial surfaces upon contact with blood, a common belief has evolved that charged nucleophilic surface groups such as amine (-NH 2 ) and hydroxyl (-OH) react with and eventually bind to the internal thioester in complement factor 3 (C3). A covalent ester or amide linkage is thereby supposed to form between C3b and the surface itself. In this report,we present complement surface binding data by null-ellipsometry for two nucleophilic surfaces (-NH 2 and -OH), for surfaces with immunoglobulin G (IgG) covalently bound, and for IgG spontaneously pre-adsorbed to hydrophobic silicon. The results reveal that the plasma proteins that were deposited during complement activation became eluted by sodium dodecyl sulfate (SDS). Hence the direct covalent binding between C3 and solid nucleophilic surfaces seems to be only of moderate importance, at least during shorter serum incubations. This strongly suggests that the prevalent covalent linkage model between solid artificial surfaces and C3b is not accurate.Instead we suggest a more pronounced role for C3 associations to other adsorbed proteins and/or electrostatic and hydrophobic protein-surface interactions.

show abstract

Section: Discussionmentioning

confidence: 99%

On the binding of complement to solid artificial surfaces in vitro

et al. 2002

View full text Add to dashboard Cite

show abstract

“…Reduced SDS PAGE (12% gels) and immunoblot analysis were performed on the eluted proteins. 61 The proteins were transferred to immobilon PVDF membranes (Millipore, Bedford, MA) and stained using a stabilized gold sol (Protogold, Cedarlane Laboratories, Hornby, Ontario, Canada). Primary antibodies to various plasma proteins, the majority of which were in the form of fractionated antisera developed in goats were used at a dilution of 1:1000.…”

Section: Sds Page and Immunoblottingmentioning

confidence: 99%

Interactions of corneal cells with transforming growth factor β2‐modified poly dimethyl siloxane surfaces

Merrett

Griffith

Deslandes

et al. 2003

J Biomedical Materials Res

View full text Add to dashboard Cite

The downgrowth of corneal epithelial cells at the interface of an artificial cornea and the host eye tissue poses a significant problem to be overcome in developing a successful implant. As a means of inhibiting the proliferation of corneal epithelial cells on the stromal surface of the implant, we examined the immobilization of transforming growth factor beta-2 (TGF-beta2) via a bifunctional poly ethylene glycol (PEG) spacer to poly dimethyl siloxane (PDMS) surfaces. Growth factor immobilization was confirmed by modification with (125)I-labeled TGF-beta 2. The modified surfaces were also characterized by advancing water contact angles, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Although the amount of growth factor covalently bound to the surface was difficult to quantify apparently due to strong interactions between the growth factor and the PEG layer and high levels of adsorption, differences in the modified surfaces, suggestive of the presence of a significant amount of TGF-beta 2, were found. In vitro interactions of the modified surfaces with human corneal epithelial and stromal cells were examined. Growth factor surface concentrations as well as culture in the absence and presence of serum and other adhesive proteins were examined. Corneal stromal and epithelial cells cultured on the TGF-beta 2-modified surfaces consistently gave results opposite to those expected. Likely, the most notable and surprising result was the almost complete lack of adhesion of the stromal cells, with coverage averaging between 3 and 5%. In comparison, corneal epithelial cell growth appeared to be promoted by the presence of the immobilized growth factor, with cell coverage averaging 50-60% at 7 days of culture. A TGF-beta 2 concentration effect was noted with both cell types in the absence of serum, with increases in the coverage at higher TGF-beta 2 concentrations. The observed cell growth appeared to be the result of interactions between the cells and active growth factor, because the addition of anti-TGF-beta 2 to the culture medium reduced cell coverage to levels similar to those noted on control surfaces. Therefore, although TGF-beta 2-modified surfaces may not be suitable as corneal epithelial cell inhibiting surfaces, interactions of surface immobilized growth factor and corneal cells are complex and should be further examined.

show abstract

“…The blots for the surfaces are much less complex than that for plasma itself, 27 suggesting some selectivity in their interactions with the plasma proteins. The gold surface shows virtually no proteins except for trace amounts of albumin, IgM, C3c, and complement factor I.…”

Section: Protein Interactionsmentioning

confidence: 99%

Peptide modified gold-coated polyurethanes as thrombin scavenging surfaces

Sun

Sheardown

Tengvall

et al. 2000

J. Biomed. Mater. Res.

Self Cite

View full text Add to dashboard Cite

Thin layers of gold were deposited on polyurethane film and chemisorbed with three peptides having an N-terminal cysteine: Cys-Pro-Arg, Cys-(L)Phe-Pro-Arg, and Cys-(D)Phe-Pro-Arg. The ability of these surfaces to act as thrombin scavengers was evaluated. The peptides are related to the known thrombin inhibitor Phe-Pro-Arg chloromethyl ketone and were shown to have significant thrombin inhibitory activity in solution. Attachment of the peptides to gold was confirmed by water contact angle and X-ray photoelectron spectroscopy measurements. Thrombin adsorption from a buffer and plasma was investigated, and chromogenic substrate assays were carried out for thrombin activity on the surfaces and in the supernatant following adsorption. The data suggest that the peptide-modified surfaces are able to adsorb thrombin with high affinity from a buffer and that thrombin is taken up selectively from plasma. The Cys-(D)Phe-Pro-Arg modified surfaces showed particularly high affinity for thrombin. It was also found that the activity of thrombin adsorbed on the peptide surfaces was inhibited, and inhibition was greatest on the Cys-(D)Phe-Pro-Arg surface. We concluded that the peptide surfaces may have potential as antithrombogenic materials via their ability to scavenge and inhibit thrombin generated as a result of blood-material contact.

show abstract

Identification of proteins adsorbed to hemodialyser membranes from heparinized plasma

Cited by 68 publications

References 34 publications

On the binding of complement to solid artificial surfaces in vitro

On the binding of complement to solid artificial surfaces in vitro

Interactions of corneal cells with transforming growth factor β2‐modified poly dimethyl siloxane surfaces

Peptide modified gold-coated polyurethanes as thrombin scavenging surfaces

Contact Info

Product

Resources

About