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Published on: October 30, 2012
A bioimaging system combining human cultured reporter cells and planar chromatography to identify novel bioactive
Ines Klingelhöfer1, Long Pham Ngoc2, Bart van der Burg2
1Institute of Nutritional Science, Chair of Food Science, and TransMIT Center for Effect-Directed Analysis, Justus Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392, Giessen, Germany.
Abstract:
For the first time, a human cancer cell line was shown to grow and be functionally active on the particulate porous adsorbent surface of separated sample mixtures. This allowed the novel combination of chromatographic separations with human cells as biological detector. As exemplary screening for cancer treatment drugs, cytotoxic substances were directly discovered in Saussurea costus and ginseng samples using the Cytotox CALUX® osteosarcoma cells (with luciferase expressing reporter gene) as detector. In addition, rosiglitazone and pioglitazone were detected as luminescent zones upon binding to the PPARγ receptor expressed in the respective CALUX cell line that was grown on the surface of the adsorbent. This demonstrates the ability to address receptor-mediated signaling with this method, and opens the perspective to use our novel bioimaging method to identify bioactive molecules targeting a wide range of pathways with toxicological, pharmaceutical and nutraceutical relevance. The new bioimaging directly pointed to individual effective compounds in multi-component mixtures. Furthermore, discovered effective compounds were directly characterized by online elution to high-resolution mass spectrometry and fragmentation.
Insights
A novel bioimaging technique uses human cancer cells on a porous surface to detect active compounds in complex mixtures. This method identifies potential cancer drugs and receptor-binding molecules, enabling direct characterization of effective compounds.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Cell Biology
Background:
- Traditional drug discovery methods face challenges in analyzing complex natural mixtures.
- Developing sensitive and specific bioassays for identifying bioactive compounds is crucial.
Purpose of the Study:
- To establish a novel bioimaging method combining chromatography with human cells as detectors.
- To demonstrate the direct discovery and characterization of bioactive molecules in complex mixtures.
Main Methods:
- Culturing human cancer cells (Cytotox CALUX® osteosarcoma cells) on a porous adsorbent surface for chromatographic separations.
- Utilizing cells expressing reporter genes (luciferase) for detecting cytotoxic and receptor-binding activities.
- Online elution coupled with high-resolution mass spectrometry and fragmentation for compound characterization.
Main Results:
- Successfully demonstrated human cancer cell growth and function on adsorbent surfaces for bioimaging.
- Identified cytotoxic substances in Saussurea costus and ginseng samples.
- Detected rosiglitazone and pioglitazone binding to PPARγ receptors.
- Directly characterized identified compounds using mass spectrometry.
Conclusions:
- The novel bioimaging method enables direct identification of bioactive compounds in multi-component mixtures.
- This technique is applicable for screening drugs, identifying toxicological compounds, and discovering pharmaceutical and nutraceutical agents.
- The method allows for addressing receptor-mediated signaling pathways and direct online characterization of active molecules.
