Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1994-08-01
1995-02-14
Spiegel, Carol A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 79, 435 792, 435 7021, 43524023, 43524027, 435204241, 435240243, 436524, 436528, 436531, 436825, 5303882, 5303911, G01N 33554
Patent
active
053895185
DESCRIPTION:
BRIEF SUMMARY
The present invention in general relates to the involvement of adhesive glycoproteins, such as vitronectin and fibronectin in the attachment and growth of animal cells on non-biological surfaces, such as polymeric plastics, metals, ceramics etc. More particularly, the present invention relates to monoclonal antibodies directed against vitronectin and fibronectin and to the use of such monoclonal antibodies in the production of a biomaterial and a device for use in in vitro cell culture. The present invention also relates to a method for analysing biological samples for the presence of adhesive glycoproteins or fragments thereof and to a method of extracting vitronectin from bovine serum or plasma, to vitronectin so produced, and to a device used in the extraction method.
One of the major difficulties presently encountered in the preparation of biomaterials for certain applications is that of ensuring cellular attachment to the biomaterial. Cellular attachment to a number of polymeric materials used for biomaterials, such as teflon, is dependent on adsorption of serum or cellular glycoproteins which are adhesive for cells, however, adsorption to these surfaces is low and non-specific. It is believed that the design of a biomaterial surface would be advantaged if glycoproteins could be specifically bound and held with higher affinity by the biomaterial surface. It would also be of advantage if the glycoproteins adsorbed onto the biomaterial surface were derived from the plasma or serum of the patient, and that the plasma or serum could be used without the requirement to initially purify the adhesive glycoproteins from the plasma or serum.
Adhesive glycoproteins such as fibronectin or vitronectin are multidomain proteins, each domain having a biological activity, and so the biological activity of a surface that has adsorbed such a glycoprotein is dependent upon the way in which the domain or domains of the glycoprotein are presented. It is believed that the ability to modify the biological activity of a surface to which adhesive glycoproteins are adsorbed by choosing the orientation in which the adhesive glycoproteins are bound would also be advantageous.
As stated previously the present invention also relates to a method of extracting vitronectin and to a device for use in such a method. There are a number of prior art methods for producing vitronectin and these methods, together with the drawbacks encountered by these methods are discussed in European patent application No. 0292663. As is stated in European patent application No. 0292663 it is known to extract vitronectin from human plasma or serum using anti-human vitronectin monoclonal antibodies. However, such methods have generally resulted in a poor recovery in the order of about 10 to 20%. The present invention enables the extraction of vitronectin from bovine plasma or serum with a recovery in the order of 90%.
The present inventors have produced two hybridoma cell lines which produce monoclonal antibodies each of which is reactive with a different epitope of bovine vitronectin. Accordingly, in a first aspect the present invention consists in a monoclonal antibody reactive with vitronectin in a manner such that when the monoclonal antibody is bound to vitronectin, the vitronectin retains its cell binding activity. The present invention further consists in a hybridoma cell line producing such a monoclonal antibody.
This monoclonal antibody has been designated A27, and the hybridomas cell line producing monoclonal antibody A27 has been designated CM 30/23 D2/G4. This hybridoma cell line has been deposited with the European Collection of Animal Cell Cultures (ECACC), Vaccine Research and Production Laboratory, Public Health Laboratory Service, Centre for Applied Microbiology and Research, Porton Down, Salisbury, Wiltshire SP4 OJG, United Kingdom, on 12 Jan. 1989 and accorded accession number 89011102.
In a second aspect the present invention consists in a monoclonal antibody reactive against bovine vitronectin in a manner such that when the monoclonal a
REFERENCES:
patent: 4059685 (1977-11-01), Johnson
patent: 4376110 (1983-03-01), David et al.
patent: 4746604 (1988-05-01), Mowshowitz
patent: 4828563 (1989-05-01), Muller-Lierheim
Harrison et al., "Adhesive responses of fibroblast and neuroblastoma cells to substrata coated with polyvalent or monoclonal antibody to fibronectin," Chem. Abs. , 99, No. 85807k (1983).
Schoen, R. et al., Monoclonal Antibodies Against Human Fibronectin Which Inhibit Cell Attachment, Hybridoma, vol. 1, No. 2, 1982 pp. 99-106 New York, US.
Kajikawa, H. et al., Characterization of Monoclonal Anitbodies Against Bovine Plasma Fibronectin, Zoological Science, vol. 5, No. 6, Dec. 1988, p. 1326, Tokyo, JP Abstract Only.
Hayman, E. et al., Serum Spreading Factor-virtonectin-is present at the Cell Surface and in Tissues, Proceedings of the National Academy of Sciences of the United States of America, vol. 80, No. 13, Jul. 1983, pp. 4003-4007, Washington, D.C., US.
Schakenraad, J. et al., Kinetics of Cell Spreading on Protein Precoated Substrata: A Study of Interacial Aspects, BIomaterials, vol. 10, No. 1, Jan. 1989, pp. 43-50, Guildford, Surrey, GB.
Salonen et al. J. Immunol. Methods V. 72: (1984) pp. 145-156.
Hashida and Ishikawa Analytical Letters vol. 18 (B9) (1985) pp. 1143-1155.
Lydon and Foulger Biomaterials vol. 9 (1988) pp. 525-527.
Cell Structure and Function, vol. 13, issued Mar. 1988, Masako Izumi, et al, `Identification of the Collagen-Binding Domain of Vitronectin Using Monoclonal Antibodies`, pp. 218-219.
Thrombosis and Haemostasis, Vo. 60, No. 3, issued Apr. 1988, Perumal Thiagarajan and Kathleen Kelly, `Interaction of Thrombin-Stimulated Platelets and Vitronectin (S-Protein of Complement) Substrate: Inhibition by a Monoclonal Antibody to Glycoprotein IIb-IIIa complex`.
Steele John G.
Underwood Patricia A.
Commonwealth Scientific and Industrial Research Organisation
Spiegel Carol A.
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