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)
The second paper is from the Hawes group within the School of Life Sciences at Oxford Brookes University where they nicely show the use of the Actin-Chromobody® in plant cells. They transferred the Chromobody in the Gateway® system and transiently expressed it inNicotiana tabacum leaves at different Agrobacterium tumefaciensconcentrations. In live cell studies they analyzed cytoskeleton dynamics (inhibition of polymerization and repolymerization) upon treatment with latrunculin B and compared the Actin Chromobody® with conventional labeling probes such as Lifeact. They concluded that Actin Chromobody® is particularly beneficial to study actin dynamics in plant cells as it does label actin without impairing dynamic movement and polymerisation of the actin filaments. (Rocchetti, A.; Hawes, C.; Kriechbaumer, V. Fluorescent labelling of the actin cytoskeleton in plants using a cameloid antibody. Plant Methods. 2014, 10. http://dx.doi.org/10.1186/1746-4811-10-12)
Last but not least the review article by Philipp Kaiser summarized the “Recent progress in generating intracellular functional antibody fragments to target and trace cellular components in living cells”. This paper gives a comprehensive overview on the application of intrabodies (e.g. nanobody, sdAb, fluorobody, Fab fragment, scFv, DARPin, monobody or fibronectin FN3) for cellular research. Among the different approaches to visualize endogenous proteins the Chromobody®technology is highlighted. Apart from High-Content Screening, other inventive applications of the Chromobody® technology are pointed out such as interfering with proteins/ protein function (e.g. HIV-1 Gag) or protein localization or induction of degradation. (Kaiser, P.D.; Maier, J.; Traenkle, B., et al. Recent progress in generating intracellular functional antibody fragments to target and trace cellular components in living cells. Biochimica et biophysica acta. 2014. http://dx.doi.org/10.1016/j.bbapap.2014.04.019)
Interested in our Lamin-Chromobody® and our Dnmt1-Chromobody®? Looking for more information on our Actin-Chromobody® and our Cell Cycle Chromobody®? Want a custom Chromobody®against a special target? Just write us an email or leave a comment below!
Posted by Kourosh on Thu, June 5, 2014