MEDINA has extensive experience in the development of different protein-based assays, from expression to purification of the protein and the development and execution of assays with a wide range of targets.
Protein Lab Services
- Pre-existing plasmids from public databases
- Cloning the gene coding for the target in a suitable expression vector (restriction sites, fusion tags)
- Expression optimization of the resulting plasmids in a set of Coli strains is performed in order to maximize the amount of soluble, fully-folded, active protein.
- A scale-up of the optimal expression condition is followed by a scouting of different lysis methods, chromatographic steps and a buffer formulation screen, to push the pure protein yield to the milligram level, so that it is ready to go for your biophysical experiments and your target-based screens.
- Buffer optimization (ionic strength, additives)
- Affinity
- Ionic exchange
- Size exclusion
- UV/Vis, Bradford quantification
- SDS PAGE
- Western Blot
- Mass spectrometry confirmation
- Development and validation of Protein-Protein Interaction screening platforms compatible with your synthetic compounds and our extracts of microbial secondary metabolites
- Know-how on Gold Standard biophysical methods for subsequent hit validation (Isothermal Titration Calorimetry, Microscale Thermophoresis)
- 96/384-well plates
- In order to complement your target-based drug discovery workflows, we develop for you Alphascreen®, Thermal Shift (Thermofluor®), HTRF®, FRET, competitive ELISA, Bimolecular Complementation (BiMC) so that you can get to your lead compounds faster.
- Target-based assays allow you to explore the unique chemical variability of MEDINA’s natural product library (see MEDINA’s Natural Product Library Section) in the High Throughput format.
- CFX96/384 Real-Time PCR Detection System (Biorad) - Thermal Shift (Thermofluor®)
- AKTA-Go (Cytiva) - Fast protein liquid chromatography (FPLC), protein purification
- Multiplate reading system (Envision, PerkinElmer) - NanoBRET, Fluorescence Polarization, HTRF®, Alphascreen®, AlphaLISA®, enzymatic degradation, ELISA
- High Content Screening System (Operetta CLS, PerkinElmer; BD Pathway 855, Becton Dickinson)
- Odyssey Infrared Imaging System (Li-COR Biosciences) – EMSA using Infrared Fluorescent Dye
