Hostname: page-component-78c5997874-m6dg7 Total loading time: 0 Render date: 2024-11-15T02:22:02.419Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and partial characterization of dogfish (Scyliorhinus canicula) antibody

Published online by Cambridge University Press:  16 October 2009

W. J. W. Morrow ,
J. E. Harris ,
D. Davies  and
A. Pulsford†
W. J. W. Morrow
Affiliation:
School of Environmental Sciences, Plymouth Polytechnic, Drake Circus, Plymouth, PL4 8AA
J. E. Harris
Affiliation:
School of Environmental Sciences, Plymouth Polytechnic, Drake Circus, Plymouth, PL4 8AA
D. Davies
Affiliation:
Department of Biological Sciences, University of Salford, Salford
A. Pulsford†*
Affiliation:
The Laboratory, Marine Biological Association, Citadel Hill, Plymouth, PL1 2PB
*
† Dr Ann Pulsford, The Laboratory, Citadel Hill, Plymouth, PL1 2PB.
Get access Rights & Permissions [Opens in a new window]

Abstract

A humoral immune response was elicited in adult dogfish by intramuscular injection of keyhole limpet haemocyanin (KLH). The antibody raised to this antigen was measured by an agglutination technique. The antibody-containing fraction of the serum was isolated in a two-step procedure utilizing gel nitration on Sepharose 6B and slab gel electrophoresis. A rabbit antiserum was then raised to the purified immunoglobulin. Further gel filtration studies revealed that the anti-KLH agglutinating activity was restricted to a highmolecular-weight serum fraction (approximately 800000) in fish immunized for up to 18 months. No anti-KLH precipitating activity was detected in fish at any stage of immunization; however immunodiffusion studies with the rabbit antiserum raised against dogfish antibody confirmed the existence of an 800000 molecular weight form and also a lighter species weighing 160000 daltons in both normal and immunized fish. The antigenic specificity of this latter molecule was not demonstrated. Immunoelectrophoretic analysis of the antibody revealed a neutral/slight anodic mobility, and after reduction and alkylation the presence of two polypeptide chains weighing 18200 and 75 860 daltons was demonstrated by thin-layer gel chromatography. These physicochemical characteristics make dogfish antibody analogous to mammalian immunoglobulin M (IgM).

Type
Research Article
Information
Journal of the Marine Biological Association of the United Kingdom , Volume 63 , Issue 2 , May 1983 , pp. 409 - 418
Copyright
Copyright © Marine Biological Association of the United Kingdom 1983

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

Clem, L. W. & Leslie, G. H., 1969. Phylogeny of immunoglobulin structure and function. In Immunology and Development (ed. Adinolfi, M.), pp. 62–84. London: Spastic International Medical Publications.Google Scholar
Corbel, M. J., 1975. The immune response in fish: a review. Journal of Fish Biology, 7, 539–563.CrossRefGoogle Scholar
Culliford, B. J., 1964. Precipitin reactions in forensic problems. Nature, London, 201, 1092–1094.CrossRefGoogle ScholarPubMed
Davies, D., 1975. Isolation and Characterization of the ‘Light’ Antibody from Atlantic Salmon. Ph.D. Thesis, University of Salford.Google Scholar
Elsen, H. M., 1974. Immunology – an Introduction to Molecular and Cellular Principles of the Immune Responses. 284 pp. Maryland, U.S.A.: Harper & Row Inc.Google Scholar
Ellis, A. E. & Parkhouse, R. M. E., 1975. Surface immunoglobulins on, the lymphocytes of the skate Raja naevus. European Journal of Immunology, 5, 726–728.CrossRefGoogle ScholarPubMed
Fänge, R. & Pulsford, A., 1983. Structural studies on lymphomyeloid tissue of the dogfish, Scyliorhinus canicula L. Cell and Tissue Research. (In the Press.)CrossRefGoogle Scholar
Gitlin, D., Perricelli, A. & Gitlin, J. D., 1973. Multiple immunoglobulin classes among sharks and their evolution. Comparative Biochemistry and Physiology, 44 B, 225–239.Google ScholarPubMed
Hale, L. J., 1965. Biological Laboratory Data, x, 147 pp. London: Methuen.Google Scholar
Mancini, G., Carbonara, A. O. & Heremans, J. F., 1965. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry, 2, 235–254.CrossRefGoogle ScholarPubMed
Miller, F. & Metzger, H., 1965. Characterization of a human macroglobulin. II. Distribution of the disulfide bonds. Journal of Biological Chemistry, 240, 4740–4745.CrossRefGoogle ScholarPubMed
Morrow, W. J. W., 1978. The Immune Response of the Dogfish Scyliorhinus canicula L. Ph.D. Thesis, Plymouth Polytechnic (CNAA).Google Scholar
Morrow, W. J. W., Harris, J. E. & Pulsford, A., 1982. Immunological responses of the dogfish (Scyliorhinus canicula L.) to cellular antigens. Acta zoologica, 63, 153–159.CrossRefGoogle Scholar
Morrow, W. J. W. & Pulsford, A., 1980. Identification of peripheral blood leucocytes of the dogfish (Scyliorhinus canicula L.) by electron microscopy. Journal of Fish Biology, 17, 461–475.CrossRefGoogle Scholar
Ouchterlony, O., 1948. In vitro method for testing the toxin producing capacity of diphtheria bacteria. Acta pathologia et microbiologica scandinavica, 25, 186–191.CrossRefGoogle ScholarPubMed
Pulsford, A., Fånge, R. & Morrow, W. J. W., 1982. Cell types and interactions in the spleen of the dogfish Scyliorhinus canicula L.: an electron microscopic study. Journal of Fish Biology, 21, 649–662.CrossRefGoogle Scholar
Radola, B. J., 1968. Thin-layer gel filtration of proteins. Journal of Chromatography, 38, 61–77.CrossRefGoogle ScholarPubMed
Rydon, H. N. & Smith, P. W. G., 1952. A new method for the detection of peptides and similar compounds on paper chromatograms. Nature, London, 169, 922–923.CrossRefGoogle Scholar
Scheidegger, J. J., 1955. Une micro-méthode de l’immunoélectrophorése. International Archives of Allergy and Applied Immunology, 7, 103–110.CrossRefGoogle Scholar
Severin, W. P. J., 1972. Latex agglutination in the diagnosis of meningococcal meningitis. Journal of Clinical Pathology, 25, 1079–1082.CrossRefGoogle ScholarPubMed
Suran, A. A., Hyde Tarail, M. & Papermaster, B. W., 1967. Immunologlobulins of the leopard shark. I. Isolation and characterization of 17S and 7S immunoglobulins with precipitating activity. Journal of Immunology, 99, 679–686.CrossRefGoogle Scholar
Voss, E. W., Russell, W. J. & Sigel, M. M., 1969. Purification and binding properties of nurse shark antibody. Biochemistry, 12, 4866–4872.CrossRefGoogle Scholar
Voss, E. W. & Siegel, M. M., 1971. Distribution of 19S and 7S antibodies during the immune response in the nurse shark. Journal of Immunology, 106, 1323–1329.CrossRefGoogle ScholarPubMed