Advanced purification platform using circularly permuted caspase‐2 for affinity fusion‐tag removal to produce native fibroblast growth factor 2

BACKGROUND Recombinant proteins produced for use as biopharmaceuticals need to harbor their native N-terminus. A drawback in expression of recombinant fusion with an affinity fusion-tag is that enzymatic or chemical processing required trim the artificial tag and release true protein interest. In many cases, however, this step generates incorrect RESULTS Human fibroblast growth factor 2 (FGF2) was expressed a Escherichia coli fed-batch cultivations. The interest (POI) carried N-terminal which enabled purification via chromatography. After removal circularly permuted human caspase-2 (cpCasp2), POI further purified using subtractive Mass spectrometric analysis confirmed authentic N-terminus POI. generated highly pure 42 ppm host cell protein, 3.7 μg mL−1 dsDNA ~ 1000 EU endotoxin. Only small number E. were co-purified Because high specificity novel protease cpCasp2, no off-target cleavage could be observed. CONCLUSION Our findings demonstrate cpCasp2 can used production fusion-protein process. This represents first case study at large laboratory scale industrially relevant protein. technology constitutes basis scalable cpCasp2-based platform process (CASPON technology) platform. © 2021 Authors. Journal Chemical Technology & Biotechnology published by John Wiley Sons Ltd on behalf Society Industry.