High-Throughput Screening Assay In 384-Well Format For The Identification Of Anti-Trypanosomal Agents Against Trypanosoma Brucei Rhodesiense And Mode Of Cell Death Study

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dc.contributor.authorLim, Kah Tee
dc.date.accessioned2019-08-01T06:21:23Z
dc.date.available2019-08-01T06:21:23Z
dc.date.issued2017-02
dc.description.abstractIn order to accelerate the discovery of novel leads for the treatment of Human African Trypanosomiasis (HAT), it is necessary to have a simple and sensitive assay to identify positive hits by whole cell viability based high-throughput screening (HTS). In this study, the HTS assay was developed in 384-well format using clear plate and black plate, for comparison. Assay robustness and reproducibility were determined under the optimized conditions in 384-well plate was well tolerated in the HTS assay, including percentage of coefficient of variation of 4.68% and 4.74%, signal-to-background ratio of 12.75 and 12.07, and Z’ factor of 0.79 and 0.82 in clear- and black-384-well plate, respectively. Final concentration of 0.30% dimethyl sulfoxide was well tolerated in the HTS assay in both types of plate. No significant differences were observed for reference drugs pentamidine and melarsoprol used in the two plate format. Therefore, clear 384-well plate was suitable for use in the HTS campaign for the identification of new compounds against Trypanosoma brucei rhodesiense. A total of 1,333 plants collected from Malaysian forest were tested using the HTS system developed and six plants were found to have potent antitrypanosomal activity. These plants extract have an IC50 value less than 1.56 μg/mL with associated selectivity indices (SI) greater than 100. Further testing against T. b. rhodesiense led to the selection of Senna spectabilis (IP 117; local name: “Kasia Kuning”) for further bioassay-guided isolation against the causative species of HAT.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/8550
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectHigh-throughput screening assay in 384-well formaten_US
dc.subjectidentification of anti-trypanosomal agentsen_US
dc.titleHigh-Throughput Screening Assay In 384-Well Format For The Identification Of Anti-Trypanosomal Agents Against Trypanosoma Brucei Rhodesiense And Mode Of Cell Death Studyen_US
dc.typeThesisen_US
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