Publication Details (including relevant citation information):
Authors: Vishal Gaurav, Susan C. Roberts
Publication date: 2009
Conference name: The American Chemical Society (ACS) Proceedings
Publisher: The ACS National Meeting, Salt Lake City, UT
Paclitaxel, an important anti-cancer drug, is a complex diterpenoid synthesized in Taxus plant cell cultures. To unravel the inherent culture heterogeneity and variability in production, our laboratory is investigating the regulation of cellular metabolism in distinct cell populations, isolated using flow cytometric cell sorting technologies. A novel indirect immunofluorescent assay was developed to detect paclitaxel accumulation in live plant cells to enable sorting of both high and low accumulating populations. This assay utilized a mouse monoclonal anti-paclitaxel antibody in conjunction with a PEconjugated goat anti-mouse secondary antibody. Concomitantly, cell viability was assessed using staining with fluorescein diacetate to ensure that stained cells were undamaged and remained intact following the procedure. Experiments were performed with variations in antibody concentrations, wash steps and incubation times to establish an optimal procedure.
Address (URL): https://sites.google.com/site/vishalpubs/Development_of_a_live_cell-based_immuno fluorescent_assay_for_the_detection_of_pharmaceutically-relevant_secondary_metab olites_in_plant_cells.pdf