Glycoengineered Recombinant Alpha1‐Antitrypsin Results in Comparable In Vitro and In Vivo Activities to Human Plasma‐Derived Protein
Frances Rocamora, Chen‐Lin Hsieh, Sanne Schoffelen, Johnny Arnsdorf, Eric A. Toth, Abdul S. Yunus, Thomas E. Cleveland, Sara Petersen Bjørn, Mina Ying Min Wu, Noel G. McElvaney, Bjørn Gunnar Rude Voldborg, Thomas R. Fuerst, Nathan E. LewisABSTRACT
Alpha‐1‐antitrypsin (A1AT) is a multifunctional, clinically important, high‐value therapeutic glycoprotein that can be used for the treatment of many diseases, such as A1AT deficiency, diabetes, graft‐versus‐host disease, cystic fibrosis, and various viral infections. Currently, the only U.S. food and drug administration‐approved treatment for A1AT disorders is intravenous augmentation therapy with human plasma‐derived A1AT (pdA1AT). In addition to its limited supply, this approach poses a risk of infection transmission, since it uses therapeutic A1AT harvested from donors. To address these issues, we sought to generate recombinant human A1AT (rhA1AT) that is comparable to its plasma‐derived counterpart using glycoengineered Chinese Hamster Ovary (geCHO‐L) cells. By perturbing nine key genes that are part of the CHO glycosylation machinery and expressing the human ST6GAL1 and A1AT genes, we obtained stable, high producing geCHO‐L lines that produced rhA1AT having a highly similar glycoprofile to pdA1AT. Additionally, the rhA1AT demonstrated in vitro activity and in vivo half‐life comparable to commercial pdA1AT. Thus, we anticipate that this platform will help produce human‐like recombinant plasma proteins, thereby providing a more sustainable and reliable source of therapeutics that are cost‐effective and better‐controlled regarding purity, clinical safety, and quality.