Vimentin dynamics can be effectively visualized using the ChromoTek Vimentin-Chromobody, e.g. to monitor epithelial-mesenchymal transition (EMT). To learn more about this unique probe attend Julia Maier’s talk “Tracing EMT with fluorescent biosensors (Chromobodies) in living lung cancer cells” on April 3, 4:20 pm in Room 144, Level 1 at AACR 2017 in Washington, DC.
Cell cycle and cell proliferation are tightly controlled cellular processes, and their deregulation is a hallmark of cancer. To enable non-invasive analysis of cell cycle in living cells, ChromoTek developed a Cell Cycle Chromobody. The Cell Cycle Chromobody (CCC) consists of a binding domain of a heavy-chain antibody highly specific against human proliferating cell nuclear antigen (PCNA), which is genetically fused to a fluorescent protein.
The ChromoTek Actin Chromobody is a live-cell probe for visualization of the actin cytoskeleton and monitoring its dynamics. The Actin Chromobody enables non-invasive labeling of actin microfilaments not only in mammalian cells, but also in cells and tissues of evolutionary distant species, such as Zebrafish (Panza et al. 2015) or plants (Rocchetti, Hawes, and Kriechbaumer 2014). The Z-stack shows confocal images of optical sections of Actin Chromobody in a Hela cell.
This time last spring we started marketing our Chromobody® plasmids. Our customers realized the impact of the technology and were immediately beginning to work with the Chromobodies®. Now, one year later the first papers are being published demonstrating the broad applicability of this intracellular fluorescent antibody technology:
The first is a paper in Molecular Cell by the Lindqvist Lab at the Karolinska Institutet in Stockholm, Sweden. They developed a FRET-based system for accurate quantification of fluorescence from single cells. By visualizing endogenous PCNA at replication foci with the Cell Cycle Chromobody® they nicely show that the mitotic entry network is linked to the completion of S phase. It does not depend on protein accumulation through G2 but is activated by mitotic phosphorylations at the end of S phase. The method they present allows analyzing live-cell as well as extracting temporal information from fixed cells based on endogenous marker proteins. (Akopyan, K.; Silva Cascales, H.; Hukasova, E., et al. Assessing kinetics from fixed cells reveals activation of the mitotic entry network at the S/G2 transition. Molecular cell. 2014, 53, 843-853. http://dx.doi.org/10.1016/j.molcel.2014.01.031)
Chromobodies® are new intracellular functional antibodies visualising dynamic changes of target proteins in live cells. With the help of NMI`s outstanding lentiviral transduction technology, Chromobodies® will be introduced into difficult-to-transfect cell lines such as primary cells. The cooperation will establish innovative cell lines available for phenotypic screening, target validation and high content analysis (HCA). The unique combination of Chromobody® nanoprobes and a comprehensive collection of disease-relevant cellular test systems will lift preclinical compound screening and pathway analysis in the early drug discovery process to a new level of performance.
ChromoTek GmbH (Munich, Germany) offers nanobody based real time assays to visualize and measure the action of candidate drugs or other substances of interest on important regulatory processes in living cells. Presently, four Chromobody® plasmids are available for analyzing the cell cycle, apoptosis, DNA methylation and cytoskeleton dynamics.
ChromoTek GmbH (Munich, Germany) and Cell Culture Service GmbH (Hamburg, Germany) have entered into a service, distribution and co-marketing partnership.
Under the agreement, Cell Culture Service will provide Chromobody® cells for direct use in live cell assays. These express fluorescently labeled intracellular antibodies allowing real time monitoring of endogenous intracellular proteins. Biotechnology or pharmaceutical companies will be offered Chromobody® cell lines or transient bulk transfections of own cell lines in assay ready formats to enhance their drug discovery and high throughput screening capabilities.
Chromobodies are single domain antibodies that are genetically fused to fluorescent proteins to serve as functional nanoprobes in living cells. The Chromobody® technology allows for the first time to trace endogenous intracellular antigens and to visualize dynamic changes of these targets in living cells. Currently available cell lines allow the high content analysis of the full cell cycle and real time monitoring of apoptotic events. Small molecule libraries can be screened for desired or adverse effects on a cellular level in multiwell formats. "The combination of dynamic live cell analysis and high content read out closes a gap in drug discovery and I am delighted to facilitate the use of this novel technology for my customers", states Dr. Oliver Wehmeier, Managing Director and President of Cell Culture Service. "We are impressed with CCS’ capabilities to manufacture any type of cell based reagent in any type of cell and assay format for Chromobody® customers " says Dr. Marion Jung, Managing Director of ChromoTek.
Customers who have licensed a cell line from ChromoTek can order these cells from CCS directly. Customers who would like to get access to ChromoTek’s technology for the first time can license the cell line or order assay-ready frozen instant cells via ChromoTek or via CCS.
About CCS Cell Culture Service (now a member of the Evotec group)
For more than eleven years, CCS has been supporting research departments of pharmaceutical and biotechnology companies in their efforts to discover new drug entities by supplying them with custom made functionally validated cell based reagents.
ChromoTek products set new benchmarks for cellular research. The company was established in 2008 as a spin-off from Munich’s Ludwig Maximilian University and is located in Martinsried, Germany’s leading biotech cluster. ChromoTek develops and markets immunological and bioimaging reagents and cellular assays for biomedical research and drug discovery. These include the GFP-Trap® for the rapid pull-down of GFP fusion proteins, GFP and RFP Booster for intensifying the fluorescence signal of GFP or RFP fusion proteins, Chromobody® based live cell assays for screening and validation and a fluorescence based protein-protein interaction assay called Fluorescent-2-Hybrid (F2H®). More than 1.900 customers from all over the world trust in ChromoTek products.
Posted by Katrin on Mar 16, 2012