Abstract 6309: Development of protein conformational array ELISA for p53 and SHP2 allosteric regulator screening and analysis

Abstract

Abstract P53 and SHP2 plays important role during cancer development. For p53, more than 50% of cancers were shown to have gene mutation in this tumor suppressor, however how these gene mutations impact the p53 protein conformation and function have not be studies in much detail. A Protein Conformational Array ELISA was developed with 17 polyclonal antibodies covering the whole p53 molecule, by measuring the linear epitope distribution on the surface of the p53 protein, the conformational (Higher Order Structure or HOS) can be described quantitively. Initial studies have shown that in lung cancer cells, the level of p53 is significantly increased and more interestingly, the p53 HOS has changed at its N-terminal as compared with the wild type. For SHP2, a protein phosphatase, a 27-antibody panel was developed to characterize the HOS changes. Because of the generic nature of the phosphate binding site, it has been very challenging to develop phosphatase inhibitors, however studies have shown that allosteric regulators can be identified with excellent anti-tumor activities. Currently, some of the allosteric regulators identified are being tested in the clinic. The SHP2 Conformational Array ELISA will provide a robust platform for the screening and testing of potential allosteric regulators with its molecular resolution and high-throughput capability. This presentation will describe the development of the two conformational arrays and their applications in the study of p53 and SHP2 from different cancer cell lines and the characterization of the allosteric regulators identified. Citation Format: Xing Wang. Development of protein conformational array ELISA for p53 and SHP2 allosteric regulator screening and analysis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6309.