Effective use of nitrocellulose-blotted antigens for phage display monoclonal antibody selection.

Francesca Bugli, Luca Masucci, Paola Cattani, Giovanni Fadda, Francesco Paroni Sterbini, Rosalia Graffeo, Raffaella Iantomasi, Riccardo Torelli, F Caridi

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

The combinatorial phage display library approach to antibody repertoire cloning offers a powerful tool for the isolation of specific antibodies to defined target antigens. Panning strategy is often a very critical point for selecting antibody displayed on the surface of bacteriophages. Most selection strategies described to date have relied on the availability of purified and often recombinant antigen, providing the possibility to perform selections on a well defined antigen source. However, when the antigen is difficult to purify by means of laborious and time-consuming chromatography procedures, panning of phage antibody libraries has to be performed on complex antigen sources such as cell surfaces or tissue sections, or even by in vivo selection methods. This provides a series of technical and experimental complications. In the present work, we successfully generated a mouse monoclonal antibody fragment from a phage display library directed against protein E7 of HPV18 avoiding antigen purification as for immunizing mice as for antibody library selection. Our work demonstrates the feasibility of phage antibody selections on antigens transferred to a nitrocellulose membrane as solid support, using one-dimensional polyacrylamide gel electrophoresis system as the only practice to separate a given antigen present in bacterial crude cell lysate.
Original languageEnglish
Pages (from-to)281-286
Number of pages6
JournalNew Microbiologica
Publication statusPublished - 2011

Keywords

  • Antibody selection
  • Human papillomavirus (HPV18)
  • Monoclonal antibody
  • Oncoprotein E7
  • Phage display

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