Abstract 2172: Quantification of ERK activity in cancer cell lysates and tumor extract using differential sensing methods

• Published in: Cancer research (Chicago, Ill.) Vol. 79; no. 13_Supplement; p. 2172
• Main Authors: Zamora-Olivares, Diana, Kaoud, Tamer S., Zeng, Lingyu, Telles, Mitchell, Anslyn, Eric V., Dalby, Kevin N.
• Format: Journal Article
• Language: English
• Published: 01.07.2019
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/1538-7445.AM2019-2172

Abstract The activity of protein kinases in biological samples is typically estimated by determining the phosphorylation status of either the kinase itself, or a known cellular substrate. This is typically evaluated by multi-step western blotting, or proteomics procedures. Such procedures typically provide qualitative estimates of the modifications in question. While immunoblotting and proteomics procedures can be used in a quantitative manner, their complicated workflow diminishes reproducibility. Futhermore, modifications do not necessarily correlate closely with a protein kinase’s activity. The motivation for this work was to assess the potential of a peptide array to quantify ERK activity in cancer cell lines and tumor samples, without the need to suppress related kinase activities. This work shows that a library of cross-reactive peptide-based biosensors, along with chemometric analysis can be used to profile a kinase activity in cancer cell lines. Significantly, the array is suitable for quantifying unknown levels of ERK activity in unfractionated cancer cell lysates and tumor samples using a multivariate regression model. The predicted values provided by our model were found to be comparable to those obtained by immune complex kinase assays. Note: This abstract was not presented at the meeting. Citation Format: Diana Zamora-Olivares, Tamer S. Kaoud, Lingyu Zeng, Mitchell Telles, Eric V. Anslyn, Kevin N. Dalby. Quantification of ERK activity in cancer cell lysates and tumor extract using differential sensing methods [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2172.