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Abstract

The chicken is a useful animal for the development of the specificantibodies against the mammalian conserved proteins. We generated twotypes of recombinant chicken monoclonal antibodies (mAbs), using a phagedisplay technique from a chicken hybridoma HUC2-13 which secreted themAb to the N-terminal of the mammalian prion protein (PrP). Althoughthe mAb HUC2-13 is a useful antibody for the prion research, thehybridoma produces a low level of antibody production. In order to producea large amount of the mAb, we have constructed a single chain fragmentvariable region (scF(V)) mAb by using the variable heavy(V(H)) and light (V(L))genes which were amplified by using the two primer pairs and theflexible linker. The two phage display mAbs (HUC2p3 and HUC2p5)expressed on a M13 filamentous phage and their soluble type mAbs(HUC2s3 and HUC2s5) were reacted with the PrP peptide antigen in theELISA. In the Western blot analysis, the mAbs HUC2p3 and HUC2s3 wereas reactive to PrP(c) from mouse brains as the mAb HUC2-13 was. The nucleotide sequences of V(H) and V(L) genes from HUC2-13 and the two cloneswere identical except for only one residue. These results indicate that themethods presented here provide an effective tool for the improvement ofthe low levels of antibody production in the chicken hybridoma system.

Designing strong inducible synthetic promoters in yeasts

“…Using the above cDNA mixtures, a scFv phage library was constructed using a previously described method 45 and used for a panning assay 46 . Here, adsorption panning used wild-type RBD to deplete scFv variants that were bound to the domain.…”

“…ELISA was performed as described previously 46 . Briefly, plate wells were coated with 50 µL (1 µg/mL) of SARS-CoV-2 Spike trimer (Cat.…”

Bioinformatics in molecular immunology laboratories demonstrated: Modeling an anti-CMV scFv antibody

“…ScFvs are commonly constructed from hybridoma, mouse immunoglobulin, sheep immunoglobulin [ 7 ], chicken immunoglobulin [ 8 , 9 ] and human antibody repertoire. [ 10 , 11 ] They are generally produced at large scales using genetically engineered cloning vectors in bacterial hosts.…”