Multispecificity of a recombinant anti‐ras monoclonal antibody

Abstract

Recombinant monoclonal antibodies (Ab's) have widespread application as research tools, diagnostic reagents and as biotherapeutics. Whilst studying the cellular molecular switch protein m‐ras, a recombinant monoclonal antibody to m‐ras was generated for use as a research tool. Antibody genes from a single rabbit B cell secreting IgG to an m‐ras specific peptide sequence were expressed in mammalian cells, and monoclonal rabbit IgG binding was characterized by ELISA and peptide array blotting. Although the monoclonal Ab was selected for specificity to m‐ras peptide, it also bound to both recombinant full‐length m‐ras and h‐ras proteins. The cross‐reactive binding of the monoclonal Ab to h‐ras was defined by peptide array blot revealing that the Ab showed preference for peptide sequences containing multiple positively charged amino acid residues. These data reinforce the concept of antibody multispecificity through multiple interactions of the Ab paratope with diverse polypeptides. They also emphasize the importance of immunogen and Ab selection processes when generating recombinant monoclonal Ab's.

Antibody Binding Epitope Mapping (AbMap) of Hundred Antibodies in a Single Run

“…In addition, our understanding of antibody multispecificity could be enhanced. According to the conventional concept, the capacity of an antibody to bind several “unrelated” targets is regarded as the multispecificity of the antibody ( 13 , 28 , 31 ). When the antibody-binding linear epitope is not available, the relationship among these “unrelated” targets could not be understood beyond their being recognized by the same antibody.…”