We successfully synthetized pure and perfectly structured Interferon Alpha-2a at 10 mg scale
Interferon Alpha-2a (INF Alpha-2a) is a type 1 interferon that consists of 165 amino acid residues, which is a therapeutic mid-size protein, is traditionnaly produced by recombinant technology. This cytokine marketed under the brandname ROFERON-A®, is extensively used for its antiviral and antineoplastic properties.
We employed a 3-fragments assembly strategy in order to ensure optimized yield and purity. Fragments were synthetized by classical Solid Phase Peptide Synthesis with our proprietary resin and purified with RP-HPLC.
The assembly of the linear protein used our proprietary technology in a one-pot reaction, without any intermediary purification. Purification of the linear protein was done by RP-HPLC.
Folding of the protein
By using modification in the protein sequence, we performed a highly efficient folding in solution, leading to high yields. After completion of the folding step, the liberation of the native folded protein was done by in-situ treatment. The final purification of the protein was performed by RP-HPLC or HIC methods.
To ensure opitmal purity and correct structure of IFN Alpha-2a had been achieved, we anlaysed this mid-size protein with the highly and reliable chromatographic and mass spectrometry methods, which we have developed..
Strong QA/QC ensured a purity in excess of 95 % of the final INF Alpha-2a.
IFN Alpha-2a is known to have 2 disulfides bridges: Cys1-Cys98 and Cys29-Cys138. We demonstrated the correct cysteines by enzymatic digestion (trypsine) and UHPLC/Mass Spectometry analysis of resulting fragments.