00000ctm a22000003a 4500 UP-1685523046126336310 Buklod 20190814112540.0 m |o d | ta 190814s2019 xx d r |||| u| (iLib)UPMNL-00016535198 ​​​​UPPharm rda eng LG 993.5 2019 P5 A46 Almario, Trystam Eymar R.​ author Investigation of the applicability of cellular thermal shift assay (CETSA) for monitoring HMG-COA reductase levels in HCT-116 Trystam Eymar R. Almario, Dethalee Gabrielle R. Velasquez ; adviser, Czarina Dominique R. Rodriguez ; co-advisers, Neil Andrew D. Bascos, Gracia Fe B. Yu. Manila Department of Pharmaceutical Chemistry, College of Pharmacy, University of the Philippines 2019. 63 leaves text rdacontent unmediated rdamedia volume rdacarrier ​Thesis (Bachelor of Science in Pharmacy)--University of the Philippines, June 2019. Yes - available to the general public. Poor success rates of drug development process may be due to lack of determination of target engagement on a more suitable assay system. CETSA is a novel technique which monitors target engagement in unmodified proteins through the measurement of thermal shift of target protein with ligand when exposed to a range of temperature. However. CETSA has no established protocols yet for screening crude extracts. With the rising popularity of organic products, researchers are driven to provide scientific explanation behind observable effects of crude extract preparations. This study aims to investigate the applicability of CETSA on screening Allium sativum crude extracts on inhibition of HMGCR. This then will elucidate the possible mechanism of A. sativum on lowering cholesterol levels. Treated HCT116 cell lysates were subjected to a range of 37-60 °C. The method of detection employed is PAGE-WB followed by visualization via chemiluminescence. A systematic troubleshooting was initiated to determine appropriate conditions to produce good CETSA results. Through this, important parameters which affects CETSA were identified namely: appropriate cell line, adequate temperature range, suitable detection system and functionality of antibodies. Given the necessary adjustments, the target protein still failed to be detected. This may be due to functionality reasons. Hence, under the tested conditions, CETSA is not applicable to monitor A. sativum crude extracts for HMGCR inhibition. For better employment of CETSA, recommendations (e.g. performing transcriptome and proteomic analysis, checking cell viability, exploring other detection systems) were asserted. Hydroxymethylglutaryl coenzyme A reductases. Cellular Thermal Shift Assay (CETSA). Velasquez, Dethalee Gabrielle R.​ author. Rodriguez, Czarina Dominique R.​ thesis adviser. Bascos, Neil Andrew D.​ thesis co-adviser. Yu, Gracia Fe B.​ thesis co-adviser. FI UP UPMNL PHARM LG 993.5 2019 P5 A46 Thesis