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Blocking Immunodominant Epitopes by Competitive Deselection
Abstract
The development of combinatorial antibody (Ab) libraries displayed on the surface of phage has led to the production of a wide range of human monoclonal antibodies (MAbs) against a plethora of viral antigens (Ag) (1-5). However, sometimes the isolation of a given Ab can be particularly difficult because of the predominance of some epitopes, and, in the case of impure Ags, because the protein or compound of interest is present in a low amount. Many techniques have been developed for overcoming this problem, including epitope-masking (6) or sandwich-capture (7) procedures. These approaches require a MAb against the Ab or epitope of interest, which is not always available. In this chapter, a new procedure of preadsorption panning is described, which facilitates the molecular cloning of MAb fragments against nonimmunodominant Ag determinants using phage-display immunoselection. The procedure, called competitive deselection, is easy, fast, inexpensive, and can be coupled with other described panning techniques; however, its chief advantage is in the isolation of Abs against less-represented Ag determinants.
Affiliation(s): (2) Istituto di Microbiologia, Facoltà di Medicina, Università di Ancona, Ancona, Italy
Series: Methods in Molecular Biology  |  Volume: 178  |  Pub. Date: Dec-28-2001  |  Page Range: 173-177  |  DOI: 10.1385/1-59259-240-6:173
Subject:  Microbiology
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