Monoclonal antibodies to plasma high density lipoprotein (HDL) of eel (Anguilla japonica)

Six week-old female mice (Balb/c) injected intraperitonealy with 50 mu g of eel high density lipoprotein (HDL) emulsified with equal volume of adjuvan t three times every two weeks. Three weeks after the third injection, hyper immunized mice were boosted by injection of 100 mu g of HDL. After 5 days, the best responding mouse to injected HDL was sacrificed, and spleen cells were fused with mouse myeloma cells (Sp2/O-Ag14), and hybridomas were cultu red in a selection medium. Monoclonal antibodies specific to apolipoprotein A-I or A-II (apoA-I or apoA-II) of HDL were obtained by cloning and reclon ing the hybridomas. Eighteen monoclonal antibodies specific to apoA-I and/o r apoApII were isolated. Antibodies in the culture medium were purified by a HiTrap Protein G or an eel-HDL column. These purified antibodies belong t o the subclass IgG1. The monoclonal antibodies specific to eel apoA-I and a poA-II secreted by clone 10D12 and 2G3, respectively, interact with serum p roteins of some fish species such as red-sea bream and carp. The anti-eel a poA-I antibody of 10D12 did not bind to serum proteins of rat, rabbit, and chicken, while the anti-eel apoA-II of 2G3 antibody did. (C) 2000 Elsevier Science Inc. All rights reserved.