Services
The high-throughput facility ROBOTEIN at CIP and SBBC offers automated screening for cloning, expression, purification and biophysical characterization on either a collaborative or service basis. European scientists can get access to Robotein through the European Commission funded Instruct Internship programme.
High-throughput cloning, mutagenesis and colony picking
Protocols have been developed for high-throughput transformation of E. coli and Pichia cells, and for site-directed or random mutagenesis. Our EasyPick robotic platform allows automated cell line generation and merges rapid selection and cloning of cells. It is well adapted to picking colonies of bacteria, yeast and eukaryotic cells.
High-throughput screening for production of recombinant proteins
Following automated colony picking, cultures in DeepWell plates allow selection of the best producers, together with screening for optimal culture medium. Expressed proteins can then be purified in a 96-well format on the central robotic workstation. Automation of this process increases throughput at each purification step, reduces human error and results in higher reproducibility with an increase in yield. Finally, the efficiency of the different purification steps can be tested rapidly on our LabChip GXII equipment.
High-throughput biophysical characterization
The combination of our central liquid handling platform with two microplate spectrophotometers is a versatile tool for the development of various high-throughput applications. At the moment, we are developing protocols for automated screening of refolding conditions (e.g. from inclusion bodies) and conformational stability measurements. Furthermore, a direct link can be created to build a protein array from hundreds of different proteins or proteins that have been submitted to different conditions. A unique combination of a high throughput protein arrayer (using 96 or 384 –well plates) and a state-of-the-art infrared imager allows hundreds of proteins to be quantitatively analysed in a few minutes for secondary structure, phosphorylation, glycosylation or any contaminant of known infrared spectrum.
High-throughput interaction technologies
The central robotic workstation is linked to an Octet HTX instrument (fortéBio), which enables real-time, label-free analysis for determination of affinity (Kd), kinetics (kon, koff) and concentration. Using microtiter plates as liquid sample holders, the system can measure 8, 16, 32, 48 or 96 samples in parallel, thus allowing optimisation of either analytical throughput or sensitivity. In the 96 biosensor mode, label-free measurement of protein concentration and kinetic characterization can be achieved at unmatched speed. In the 8 and 16 biosensor modes, high sensitivity allows measurement of small molecule binding interactions and protein quantitation down to 50 ng/mL.
High-throughput enzymatic assays
The central robotic workstation is linked to two microplate spectrophotometers that allow UV/Vis absorbance and fluorescence measurements, together with chemiluminescence detection in 96- or 384-well plates. These instruments can be used in combination for development of easy, fast and robust enzymatic assays. For instance, we have performed automated quantification of carbohydrate-isomerases, several dehydrogenases and various β-lactamases.
