Protein Lab

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
  • 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