Characterizing monoclonal antibody epitopes by filtered gene fragment phage display

Roberto Di Niro, Fortunato Ferrara, Tarcisio Not, Andrew R.M. Bradbury, Fernando Chirdo, Roberto Marzari, Daniele Sblattero

Research output: Contribution to journalArticlepeer-review

Abstract

In the present paper, we describe a novel approach to map monoclonal antibody epitopes, using three new monoclonal antibodies that recognize h-TG2 (human transglutaminase 2) as an example. The target gene was fragmented and cloned upstream of an antibiotic-resistance gene, in the vector pPAO2, to select for in-frame polypeptides. After removal of the antibiotic-resistance gene by Cre/Lox recombination, an antigen fragment phage display library was created and selected against specific monoclonal antibodies. Using the h-TG2 fragment library, we were able to identify epitopes. This technique can also be broadly applied to the study of protein-protein interactions.

Original languageEnglish
Pages (from-to)889-894
Number of pages6
JournalBiochemical Journal
Volume388
Issue number3
DOIs
Publication statusPublished - 15 Jun 2005
Externally publishedYes

Keywords

  • Epitope mapping
  • Phage display
  • Transglutaminase
  • β-lactamase

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