Quantifying the coverage of the human proteome with renewable antibodies.

Antibodies are critical reagents to detect and characterize human proteins. The biomedical research community aspires to have at least one potent and selective renewable antibody for each human protein, and for each of the most common applications. To quantify progress toward this goal, a standardized characterization approach to assess the performance of 614 commercial antibodies for 65 neuroscience-related proteins in Western blot, immunoprecipitation, and immunofluorescence applications, using isogenic knockout cells as controls. Side-by-side comparisons of the antibodies demonstrated that: i) for each application tested, approximately half of this protein set was covered by at least one high-performing antibody, suggesting that coverage of human proteins by commercial antibodies is significant but gaps are still present; ii) on average recombinant antibodies performed better than monoclonal or polyclonal antibodies; iii) hundreds of underperforming antibodies have been used in published articles. Encouragingly, more than half of the commercial antibodies that did not perform as expected were removed from the market by manufacturers or had alterations made to their recommended usage based on this body of work. This work suggests an efficient strategy toward achieving coverage of the human proteome with selective renewable antibodies is to mine the existing commercial antibody repertoire, using this knowledge to focus new renewable antibody generation efforts.