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Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus
Author(s) -
Soonthonsrima Tanapon,
Wangman Pradit,
Chaivisuthangkura Parin,
Pengsuk Chalinan,
Sithigorngul Paisarn,
Longyant Siwaporn
Publication year - 2019
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/are.13894
Subject(s) - biology , nile tilapia , oreochromis , antiserum , antibody , monoclonal antibody , tilapia , microbiology and biotechnology , bovine serum albumin , western blot , immune system , immunoglobulin m , immunoglobulin g , virology , fishery , immunology , biochemistry , fish <actinopterygii> , gene
The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer , as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.