Fine epitope specificity of antibodies against interleukin-2 explains their paradoxical immunomodulatory effects

Delineating the functional map of the interaction between nimotuzumab and the epidermal growth factor receptor

“…been associated to recognition of particular epitopes, 29,30 and to the creation or disruption of different interactions. 31,32 The identity of the epitopes can be deciphered through either structural 33 or functional methods.…”

“…Successful display of EGFR Dom III +482-514 (a complex protein fragment having five disulfide bonds) extended the scope of previous experiences involving interleukin-2 as the target. 29,30 Despite these and other examples of display of complex mammalian proteins on filamentous phages, 39 the inability to display a given target in an appropriate antigenic form would preclude our mapping strategy to be used as described here. Yeast or ribosome display could be valuable alternatives in such cases.…”

High throughput functional epitope mapping: Revisiting phage display platform to scan target antigen surface

“…While lowlevel random diversification of phagemid-cloned Ag gene by error-prone PCR was initially performed, 25,26 resembling the usual yeast display-based mapping procedures, detailed information about the residues contributing to epitope formation has been recently obtained through comprehensive mutagenesis scanning of the role of solvent-exposed side chains within a discrete surface patch. [5][6][7][8] Once the target Ag is phage-displayed and a putative relevant antigenic region within it is identified through any available method, focused exploration of such region can result in a very high local sequence space coverage (Fig. 1).…”

“…While full-length interleukin-2 (IL-2), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) have been displayed on filamentous phages for mapping purposes, [5][6][7]9 restricting the scanning to the individual domain containing the epitope(s) under investigation is the alternative of choice for larger multi-domain Ags, as it has been done with phage-displayed EGF receptor (EGFR) domain III. 8 In any case, the original folding/antigenicity of the molecule must be preserved.…”

“…The current article summarizes mapping experiences accumulated during the last 2 y on the bases of phage display of native Ags and a comprehensive mutagenesis exploration of their surfaces to locate the target epitopes. [5][6][7][8][9] The usefulness of such an approach in the context of a plethora of available epitope discovery techniques is discussed. Despite the wide use of phage display to study protein interactions, including epitope mapping, 10 the particular strategy we review here had not been fully exploited until recently.…”