Monoclonal Antibodies against Specific p53 Hotspot Mutants as Potential Tools for Precision Medicine

Abstract

Summary: The large number of mutations identified across all cancers represents an untapped reservoir of targets that can be useful for therapeutic targeting if highly selective, mutation-specific reagents are available. We report here our attempt to generate such reagents: monoclonal antibodies against the most common R175H, R248Q, and R273H hotspot mutants of the tumor suppressor p53. These antibodies recognize their intended specific alterations without any cross-reactivity against wild-type (WT) p53 or other p53 mutants, including at the same position (as exemplified by anti-R248Q antibody, which does not recognize the R248W mutation), evaluated by direct immunoblotting, immunoprecipitation, and immunofluorescence methods on transfected and endogenous proteins. Moreover, their clinical utility to diagnose the presence of specific p53 mutants in human tumor microarrays by immunohistochemistry is also shown. Together, the data demonstrate that antibodies against specific single-amino-acid alterations can be generated reproducibly and highlight their utility, which could potentially be extended to therapeutic settings. : Hwang et al. generate mutation-specific monoclonal antibodies with high sensitivity and specificity against three of the most common p53 hotspot mutations. These reagents represent the next generation of antibodies against single-amino-acid alterations, which could have potential in the diagnosis and therapeutic targeting of the many alterations found in disease states. Keywords: amino acid-specific antibody, diagnosis, hotspot mutations, mAbs, mutation specific, p53, precision medicine, therapeutic antibody, tumor suppressor