Construction of recombinant monoclonal antibodies from a chicken hybridomaline secreting specific antibody

The chicken is a useful animal for the development of the specific antibodi es against the mammalian conserved proteins. We generated two types of reco mbinant chicken monoclonal antibodies (mAbs), using a phage display techniq ue from a chicken hybridoma HUC2-13 which secreted the mAb to the N-termina l of the mammalian prion protein (PrP). Although the mAb HUC2-13 is a usefu l antibody for the prion research, the hybridoma produces a low level of an tibody production. In order to produce a large amount of the mAb, we have c onstructed a single chain fragment variable region (scF(V)) mAb by using th e variable heavy (V-H) and light (V-L) genes which were amplified by using the two primer pairs and the flexible linker. The two phage display mAbs (H UC2p3 and HUC2p5) expressed on a M13 filamentous phage and their soluble ty pe mAbs (HUC2s3 and HUC2s5) were reacted with the PrP peptide antigen in th e ELISA. In the Western blot analysis, the mAbs HUC2p3 and HUC2s3 were as r eactive to PrPc from mouse brains as the mAb HUC2-13 was. The nucleotide se quences of V-H and V-L genes from HUC2-13 and the two clones were identical except for only one residue. These results indicate that the methods prese nted here provide an effective tool for the improvement of the low levels o f antibody production in the chicken hybridoma system.