Automated system for the detection, imaging and storage of protein crystal images grown under controlled temperature, ROCK IMAGER®
The facility has a variety of commercial crystallisation screening kits and incubators at 10 °C (Raypa FKS 1800) and 18 °C in a dedicated, temperature controlled room. Crystallization plates are set with an Oryx4 crystallization robot which provides the opportunity of screening a great range of crystallization conditions saving time and sample quantity (https://www.douglas.co.uk/oryx4.htm). Crystallisation conditions can be screened in 96-well plates (SWISSCI preferably, sitting drops only) in hundreds of nonoliter volumes. Screening for sitting drop experiments is carried out using plates that are prefilled with reservoir solutions by the users (reservoirs can be filled manually with a 8-channel pipette). Crystallisation trials are monitored with a Leica 12.5M stereoscope.
The facility has a variety of commercial crystallisation screening kits and incubators at 4 °C (Fiocchetti Medica 300) and 18 °C (FOC225i, VELP Scientifica). Crystallisation conditions can be screened either in 96-well plates or in 24-well plates in milliliter droplets. Crystallisation trials are monitored, visualized over time with Euromex Z-range by Euromex and Zeiss™ Stemi 2000-C & SV6 Stereo Microscopes and crystal images of high quality can be obtained with digital Canon camera.
Visit the provider's website: http://inspired.aua.gr/instruct/
Euromex Z-range zoom stereo microscope equipped with Cold light source LE.5210 apparatus (Euromex) and digital camera canon EOS3500.
The Douglas OryXNano crystallization robot provides the opportunity of screening a great range of crystallization conditions saving time and sample quantity. Oryx system uses multi-bore dispensing tips (microtips), which have several independent channels to dispense small volumes. At the end of a 3-bore tip there are three holes. Each channel dispenses a different solution. The solutions do not mix in the tip - they mix in the drop after they are dispensed. This means that there is no dead-volume.The microtip always touches the plate when liquids are dispensed. This gives very reliable dispensing. Stock solutions do not come into contact with the motorized syringes, which are filled with degassed pure water - this avoids the need for flushing out the syringes when the stock solutions are changed.The manual syringes on the front panel are used to refill the motorized syringes, to remove air bubbles, and to load stock solutions for optimization experiments.Screening for sitting drop is carried out using plates that are prefilled with reservoir solutions. (Reservoirs can be filled manually with a 12-channel pipette. A very simple user-interface is used to design experiments, as shown below. The correct plate can be selected from a database of all well-known crystallization plates (if your favorite isn't there, let us know!), the volumes of each ingredient of a drop are entered in a simple form. Individual wells can be selected or skipped by clicking on the plate with a mouse.
Automated system for the detection, imaging and storage of protein crystal images grown under controlled temperature, ROCK IMAGER®
Professor Elias Eliopoulos is the Director of the Genetics Laboratory at the Biotechnology Department of the Agricultural University of Athens (A.U.A.) and leads the bioinformatics and structural biology group. He is professor of Biochemistry, a biophysicist and crystallographer by training with considerable experience in biomolecular structure analysis, epitope mapping of protein receptors, genetic polymorphisms and variation, cloning, with experimental and computational methods, protein structure prediction, ligand and drug design, protein design, in silico antibody design and biosoftware development. The group has experience and international reputation on computational protein folding, ab initio and homology modeling of proteins derived from edge gene research and membrane protein modeling. The group is also involved in many ligand computational design projects for pharmaceutical, medicinal and agricultural applications.
Euromex Z-range zoom stereo microscope equipped with Cold light source LE.5210 apparatus (Euromex) and digital camera canon EOS3500.
EDUCATION
2019: PhD in Structural Biology and Protein Biomaterials (UoC, IMBB-FORTH - Greece)
2012: M.Sc. in Protein Biotechnology (UoC-Greece)
2009: Diploma Degree in Biology (UoC-Greece)
WORKING EXPERIENCE
Oct 2019 – Now: Post-Doctoral Researcher at I.M.B.B.-F.O.R.TH, Greece
(Laboratory of Structural Biology)
Sep 2020 – Now: Autonomous teaching and exam organizing of the under-graduate courses
"Enzymatic Biotechnology" and “Protein Engineering” at the Dept. of
Biology of the Uni. of Crete
2014 – 2020: Experienced scientist position at I.M.B.B.-F.O.R.TH, Greece (InnovCrete project /FP7-Regpot program) and at University of Crete
Oct. 2010 –Jan. 2011: Laboratory Teaching Assistant of Microbiology Teaching Lab
at the Dept. of Biology, Uni. of Crete
SELECTED PUBLICATIONS
*Kefala K., Kotsifaki D., Providaki M., Kapetaniou E., Rahme L. and Kokkinidis M. Purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal fragment of the MvfR protein of Pseudomonas aeruginosa. (2012). Acta. Cryst F, 68, 695-697
*Kefala A., Kotsifaki D., Providaki M., Amprazi M. and Kokkinidis M. Expression, purification and crystallization of a protein resulting from the inversion of the aminoacid sequence of a helical bundle. (2017). ). Acta. Cryst F, 73, 51-53
Dimovasili C., Fadouloglou V.E., Kefala A., Providaki M., Kotsifaki D, Sarrou I., Plaitakis A., Zaganas I. and Kokkinidis M. Crystal structure of glutamate dehydrogenase 2, a positively selected novel human enzyme involved in brain biology and cancer pathophysiology. Journal of Neurochemistry (accepted)
RESEARCH PROFILE
Being in a Structural Biology group since 2011, Dr. Kefala A. has studied deeply the protein folding problem, elucidating the relation between amino acid sequence and structural properties of α-helical bundles. The knowledge obtained was used for protein design leading to bio-derived materials for bio-medical applications and basic research through structural studies of protein folding through reverse sequences.
Notable achievements include:
The elucidation of the protein folding of mutants of the α-helical protein Rop that is used as a canonical paradigm of this structural motif.
Protein design of novel protein-based biomaterials with specific engineered properties.