Hostname: page-component-78c5997874-xbtfd Total loading time: 0 Render date: 2024-11-05T08:39:38.315Z Has data issue: false hasContentIssue false
  • English
  • Français

A New Enzyme Immunoassay for Activation of Platelets by Biomaterials: Reduced Activation by Phosphorylcholine-Coated Surfaces

Published online by Cambridge University Press:  15 February 2011

Ewan J. Campbell ,
Martin C. Wiles ,
Roger R. C. New  and
Stephen A. Charles
Ewan J. Campbell
Affiliation:
Biocompatibles Limited, Brunel Science Park, Kingston Lane, Uxbridge, Middlesex, UB8 3PQ, United Kingdom
Martin C. Wiles
Affiliation:
Biocompatibles Limited, Brunel Science Park, Kingston Lane, Uxbridge, Middlesex, UB8 3PQ, United Kingdom
Roger R. C. New
Affiliation:
Biocompatibles Limited, Brunel Science Park, Kingston Lane, Uxbridge, Middlesex, UB8 3PQ, United Kingdom
Stephen A. Charles
Affiliation:
Biocompatibles Limited, Brunel Science Park, Kingston Lane, Uxbridge, Middlesex, UB8 3PQ, United Kingdom
Get access

Abstract

An enzyme immunoassay has been developed to semi-quantitate the degree of activation of platelets adhering to biomaterials. The method employs a monoclonal mouse anti-GMP-140 antibody. The glycoprotein GMP-140 is contained in alpha granules of resting platelets and is translocated to the outer membrane of platelets during activation via the thrombin-dependent pathway. A polyclonal anti-mouse IgG peroxidase conjugate is then added and platelet activation can be semi-quantitated by use of a suitable substrate, such as OPD.

The results obtained by this method compare well with scanning electron microscopy, and are shown to be less prone to the misinterpretation often associated with other conventional assay systems such as the determination of platelet adhesion by detection of ATP.

The adhesion and activation of platelets by biomaterials has been compared before and after coating these materials with phosphorylcholine (PC) derivatives. PC, in its form as a headgroup in membrane phosphoglyceride (lecithin) and ceramide (sphingomyelin) is a ubiquitous component of cell membranes, and displays extremely low interaction and binding with plasma proteins such as Factor XII [1,2], complement [2], fibrinogen [3,2], immunoglobulins [2] and albumin [2,4].

Using the immunoassay described here activation of platelets is seen to be dramatically reduced after coating surfaces, such as polyethylene, with PC compared with the untreated control. A PCcoated surface should therefore be an ideal starting point for developing tissue-inducing biomaterials. In addition endothelial cells contain GMP-140 in their secretory granules [5]. This would make this assay a potential tool in the investigation of endothelial cell - biomaterial interaction.

Keywords

HaemocompatibilityPlatelet ActivationGMP-140Di-Acetylenic Phosphorylcholine
Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 252: Symposium T – Tissue-Inducing Biomaterials , 1991 , 229
Copyright
Copyright © Materials Research Society 1992

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

[1] Campbell, E.J., Boyd, T.A., Murphy, G.C., & Yianni, Y.P. Assessment of the Haemocompatibility of Biomimetic Materials. In programme, Kunststoffe in der Medizin, IV. Kolloquium uiber Biomaterialien. 1990, Aachen.Google Scholar
[2] Ishihara, K., Ziats, P.N., Tierney, B.P., et al. Protein adsorption from Human Plasma is Reduced on Phospholipid polymers. J. Biomedical Materials Research, 25, 1991:1397–1407 Google Scholar
[3] Campbell, E.J., Sullivan, A.M., Pearce, D.J., New, R.R.C & Charles, S.A. Correlation between antibody available protein and platelet adhesion on biomaterials and biomimetic surfaces. In programme, Kunststoffe in der Medizin, V. Kolloquium über Biomaterialien. 1991, Aachen.Google Scholar
[4] Campbell, E.J., Sullivan, A.M., New, R.R.C & Charles, S.A.. Biotin labelled albumin - a simple method for the study of protein adsorption on a phosphorylcholine surface. (Abstract) In: Programme, ‘9th European Conference on Biomaterials’, 1991 Chester: ESB p147.Google Scholar
[5] Mc.Ever, R.P. Properties of GMP-140, an induceable Granule Membrane Protein of Platelets and Endothelium. Blood Cells. 16, 1990,:73–83.Google Scholar
[6] Sheppeck, R.A., Bentz, M., Dickson, C., et. al. Examination of the Roles of Glycoprotein lb and Glycoprotein IUb/Ifla in Platelet Deposition on an Artificial Surface Using Clinical Antiplatelet Agents and Monoclonal Antibody Blockade. Blood 78, No. 3, 1991: 673–680.Google Scholar
[7] Cox, A.D. and Goodall, A.H. Activation-specific Neo-Antigens on Platelets Detected by Monoclonal Antibodies. Current Studies in Haematology and Blood Transfusion, Albertini, Lenfant, Mannucci & Sixma (Eds.), Pub Kager, Basel, Switzerland, 1991, No 58,: 194–199.Google Scholar
[8] Hsu-Lin, S-C, Berman, C.L., Furie, B.C., et.al. A platelet membrane glycoprotein expressed during platelet activation and secretion: Studies using a monoclonal antibody specific for thrombinactivated platelets. J Biol Chem 259, 1984: 9121–9126.Google Scholar
[9] McEver, R.P. and Martin, M.N., A monoclon antibody to a membrane glycoprotein binds only to activated platelets. J. BiolChem 259, 1984: 9799–9804.Google Scholar
[10] Sigelman, M., Sweetening the selection pot. Current Biology, 1, No.2, 1991,: 125–128.Google Scholar
[11] Larsen, E., Celi, A., Gilbert, G.E., et. al. PADGEM Protein: A Receptor That Mediates the Interaction of Activated Platelets with Neutrophils and Monocytes. Cell 59, 1989: 305–312.Google Scholar
[12] Exon Chemical Limited, Arundel Towers, Portland Terrace, Southampton, S09 2GW, U.K.Google Scholar
[13] Chapman, D. 1979 European patent 32622.Google Scholar
[14] Polymer Products (U.K), Du Pont (U.K.) Ltd., Maylands Avenue, Hemel Hempstead, Hertfordshire, HT2 7DP, United Kingdom.Google Scholar
[15] Immunotech, supplied by The Binding Site, University of Birmingham, Research Institute, 97, Vincent Drive, Edgbaston, Birmingham, U.K.Google Scholar
[16] Sigma Chemical Company Ltd., Fancy Road, Poole, Dorset, BH17 7TG.Google Scholar
[17] Bio-orbit (UK) Limited, Wessex House, 127 High Street, Hungerford, Berkshire, RG17 ODL.Google Scholar
[18] Zucker, M.B. & Peerschke. Specific binding of fibrinogen to platelets: Relationship to shape change and aggregation. 1980 In Platelets: Cellular response mechanisms and their biological significance. Ed Rotman, Mayer, Gitler and Silberberg. Pub: John Wiley & Sons, Chichester.: 157–168.Google Scholar
[19] Hirudin, Markwardt, F. and derivatives as anticoagulant agents. Thrombosis and Haemostasis 66, 1991: 141–152.Google Scholar
[20] Wako Chemicals GmbH, Nisdanstr. 2, 4040 Neuss 1, Germany.Google Scholar