Investigation of physiological mechanisms at a cellular level often requires production of high quality antibodies, frequently using synthetic peptides as immunogens. Here we describe a new, web-based software tool called NHLBI-AbDesigner that allows the user to visualize the information needed to choose optimal peptide sequences for peptide-directed antibody production (http://helixweb.nih.gov/AbDesigner/). The choice of an immunizing peptide is generally based on a need to optimize immunogenicity, antibody specificity, multi-species conservation, and robustness in the face of post-translational modifications (PTMs). AbDesigner displays information relevant to these criteria as follows: 1) Immunogenicity Score, based on hydropathy and secondary structure prediction; 2) Uniqueness Score, a predictor of specificity of an antibody against all proteins expressed in the same species; 3) Conservation Score, a predictor of ability of the antibody to recognize orthologs in other animal species; and 4) Protein Features that show structural domains, variable regions, and annotated PTMs that may affect antibody performance. AbDesigner displays the information online in an interactive graphical user interface, which allows the user to recognize the trade-offs that exist for alternative synthetic peptide choices and to choose the one that is best for a proposed application. Several examples of the use of AbDesigner for the display of such trade-offs are presented, including production of a new antibody to Slc9a3. We also used the program in large-scale mode to create a database listing the 15-amino acid peptides with the highest Immunogenicity Scores for all known proteins in five animal species, one plant species (A. thaliana), and S. cerevisiae.
- post-translational modification
- Copyright © 2011, American Journal of Physiology - Cell Physiology