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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.
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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

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