Selection of human antibody fragments on the basis of stabilization of the variable domain in the presence of target antigens

Hideki Watanabe, Kouhei Tsumoto, Ryutaro Asano, Yoshiyuki Nishimiya, Izumi Kumagai

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Here we report a novel method for selecting human antibody fragments from nonimmunized variable domain libraries. The antibody fragments are selected on the basis of stabilization of the variable domain fragment (Fv) in the presence of target antigens ("open sandwich selection"). One variable domain is displayed on phages and another is prepared as soluble molecules. These two reagents are mixed with the biotinylated target molecule and ternary complexes are captured by using streptavidin-conjugated magnet beads. After extensive washing, enriched clones are eluted by using target antigen. Some of the clones selected after 3 rounds are prepared as soluble domains, which then undergo another selection process. We obtained several human antibody fragments specific for human soluble erythropoietin receptor by using this method. Our method minimizes several of the disadvantages associated with human antibody selection through a phage-display system, such as construction of a large-scale library, deletion of genes during selection, and nonspecific binding.

Original languageEnglish
Pages (from-to)31-36
Number of pages6
JournalBiochemical and biophysical research communications
Volume295
Issue number1
DOIs
Publication statusPublished - 2002

Keywords

  • Erythropoietin receptor
  • F
  • Human antibody
  • In vitro selection
  • Phage display

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Selection of human antibody fragments on the basis of stabilization of the variable domain in the presence of target antigens'. Together they form a unique fingerprint.

Cite this