Construction of recombinant monoclonal antibodies from a chicken hybridoma line secreting specific antibody

Naoto Nakamura, Yuri Aoki, Hiroyuki Horiuchi, Shuichi Furusawa, Hachiro I. Yamanaka, Tetsuyuki Kitamoto, Haruo Matsuda

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

The chicken is a useful animal for the development of the specific antibodies against the mammalian conserved proteins. We generated two types of recombinant chicken monoclonal antibodies (mAbs), using a phage display technique from a chicken hybridoma HUC2-13 which secreted the mAb to the N-terminal of the mammalian prion protein (PrP). Although the mAb HUC2-13 is a useful antibody for the prion research, the hybridoma produces a low level of antibody production. In order to produce a large amount of the mAb, we have constructed a single chain fragment variable 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 the flexible 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 the ELISA. In the Western blot analysis, the mAbs HUC2p3 and HUC2s3 were as 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 clones were identical except for only one residue. These results indicate that the methods presented here provide an effective tool for the improvement of the low levels of antibody production in the chicken hybridoma system.

Original languageEnglish
Pages (from-to)191-198
Number of pages8
JournalCytotechnology
Volume32
Issue number3
DOIs
Publication statusPublished - 2000

Keywords

  • Chicken
  • Immunoglobulin gene
  • Monoclonal antibody
  • Phage display
  • Prion protein
  • Recombinant antibody

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

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