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PhytomicsQC™ |
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| Genomic Bioreponse Profiling |
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| While LC/MS represents one of the most powerful chemical analysis methodologies, there is no single analysis system that is capable of detecting every type of phytochemical compound. Hence, a separate and orthogonal methodology is required that complements the strengths of chemical characterization. A powerful and novel component of the PhytomicsQC™ platform is to characterize not only the phytochemical compound pattern but also the resulting biological response to this collection of phytochemicals. |
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| Using a living human cell line and the genomic response transcription profile (transcriptome) as a sensitive detector, one can define an objective, unique subset of the expressed genes that defines a signature QC pattern for each botanical. A typical signature set will involve between 20 and 40 genes that can be developed into a routine plate-based quantitative real time-PCR (qRT-PCR) assay. Analysis of the expression profiles of different herbal formulations demonstrate that the response genes that are differentially expressed form a unique and quantitative set for each of the botanicals. This bioresponse gene pattern can be quantified and statistically compared using the PhytoVeiwerBR™ software analysis tools for the purpose of QC. In taking this approach, PhytoCeutica has combined the use of commercially available gene chips for initial screening with a statistical and informatics approach for candidate and signature gene set selection, plus real-time qRT-PCR for confirmation and final assay development. This strategy is outlined in Figure 2. |
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| Gene expression profiling offers several advantages: (1) It is a global and general bioassay, capable of integrating the bioeffects of the entire spectrum of phytochemicals, including small molecules and macromolecules; (2) gene arrays and qRT-PCR are mature, robust, and accepted technologies; and (3) each botanical has a unique and diagnostic gene regulation pattern. |
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| Figure 2. Workflow for the expression profiling and unique gene pattern identification of a botanical extract. Affymetrix microarray chips are used to generate the profile of transcriptional regulation of over 30,000 genes for the response of the cell line to a particular botanical. The collection of regulated genes is compared with the botanical database and statistical algorithms are used to select a candidate bioresponse gene that can then be validated and refined using quantitative RT-PCR to a small, unique set of signature genes. |
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