Generation of a mammalian stable cell line and purification of a recombinant protein of interest

In order to establish a proof of concept using an animal model, a client approached us in order to produce a recombinant protein of interest in the magnitude of tenths of milligrams. Based on the protein characteristics, a mammalian system was chosen for the production. Under the prescriptive of a future cGMP production, following the current guidelines, areta cloned the protein of interest in a vector suitable for high level expression in mammalian cells.

areta has generated a stable cell line that was adapted for growth in suspension and in serum free medium. After a literature search and a technology transfer, we have set-up the downstream processing implementing a tangential flow filtration for the concentration of the harvest and established an efficient and robust process for the purification of the protein of interest. Before delivery to the client, the protein batch was subjected to a comprehensive QC testing and product characterization.

Generation of monoclonal antibodies and development of an ELISA kit for the quantification of a small biomolecule

A client needed an ELISA method for the detection and quantification of a molecule in biological samples. Starting from the hapten molecule, we coupled it to a carrier protein after chemical priming. After the immunization program, we have generated a panel of monoclonal antibodies that were combined and tested as capture and detection antibodies in order to obtain the best pair.

The detection antibody was coupled to different molecules to test different detection systems. The optimal dilutions were determined by checkboard titration assay and once the conditions were defined, aliquots of small working volumes were prepared. The ELISA test was characterized for the reproducibility, linearity, sensitivity, precision and recovery.