eCite Digital Repository

Production of polyclonal antisera against barramundi (Lates calcarifer Bloch) serum immunoglobulin derived from affinity columns containing mannan-binding protein or staphylococcal protein A

Citation

Crosbie, PBB and Nowak, BF, Production of polyclonal antisera against barramundi (Lates calcarifer Bloch) serum immunoglobulin derived from affinity columns containing mannan-binding protein or staphylococcal protein A, Aquaculture, 211, (1) pp. 49-63. ISSN 0044-8486 (2002) [Refereed Article]

DOI: doi:10.1016/S0044-8486(02)00136-9

Abstract

Barramundi (Lates calcarifer Bloch) immunoglobulin (Ig) was purified by affinity chromatography using staphylococcal protein A (SpA) or mannan-binding protein (MBP) as capture ligands. Both ligands realised a pure product, although the SpA column resulted in a higher yield. In common with other teleosts, barramundi Ig appears to be a tetramer made up of two heavy chain (HC) and two light chain (LC) moieties with individual molecular weights (MW), determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), of 24 and 86 kDa, respectively. Gel filtration indicated that the native Ig molecule had a molecular weight of approximately 929 kDa. Western blot analyses demonstrated that rabbit antiserum raised against the whole MBP fraction was specific for HC and LC Ig components, whereas antisera directed against the HC or LC constituents of the SpA fraction were specific for HC and LC components, respectively. Analyses by flow cytometry and enzyme-linked immunosorbent assay (ELISA) revealed reactivity of all antisera with varying percentages of barramundi lymphocytes and SpA-purified barramundi Ig, respectively. © 2002 Elsevier Science B.V. All rights reserved.

Item Details

Item Type:Refereed Article
Research Division:Agricultural and Veterinary Sciences
Research Group:Fisheries Sciences
Research Field:Fish Pests and Diseases
Objective Division:Animal Production and Animal Primary Products
Objective Group:Fisheries - Aquaculture
Objective Field:Fisheries - Aquaculture not elsewhere classified
Author:Crosbie, PBB (Dr Philip Crosbie)
Author:Nowak, BF (Professor Barbara Nowak)
ID Code:23928
Year Published:2002
Web of Science® Times Cited:14
Deposited By:TAFI - Aquaculture
Deposited On:2002-08-01
Last Modified:2003-05-28
Downloads:0

Repository Staff Only: item control page