Isolation and partial characterization of dogfish (Scyliorhinus canicula) antibody
Morrow, W.J.W. and Harris, J.E. and Davies, D. and Pulsford, A. (1983) Isolation and partial characterization of dogfish (Scyliorhinus canicula) antibody. Journal of the Marine Biological Association of the United Kingdom, 63 (2). pp. 409-418.
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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 ofthe serum was isolated in a two-step procedure utilizing gel filtration 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 to18 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 75860 daltons was demonstrated by thin-layer gel chromatography. These physicochemical characteristics make dogfish antibody analogous to mammalian immunoglobulin M (IgM).
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