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Influence of carbohydrates on the stability and structure of a hyperglycosylated human interferon alpha mutein


, : Influence of carbohydrates on the stability and structure of a hyperglycosylated human interferon alpha mutein. Biochimie 92(8): 971-978

Protein physical and chemical instability is one of the major challenges in the development of biopharmaceuticals during every step of the process, ranging from production to final delivery. This is particularly applicable to human recombinant interferon alpha-2b (rhIFN-alpha2b), a pleiotropic cytokine currently used worldwide for the treatment of various cancer and chronic viral diseases, which presents a poor stability in solution. In previous studies, we have demonstrated that the introduction of four N-glycosylation sites in order to construct a heavily glycosylated IFN variant (4N-IFN) resulted in a markedly prolonged plasma half-life which was reflected in an enhanced therapeutic activity in mice in comparison with the commercial non-glycosylated rhIFN-alpha2b (NG-IFN). Herein, we evaluated the influence of glycosylation on the in vitro stability of 4N-IFN towards different environmental conditions. Interestingly, the hyperglycosylated cytokine showed enhanced stability against thermal stress, acid pH and repetitive freeze-thawing cycles in comparison with NG-IFN. Besides, microcalorimetric analysis indicated a much higher melting temperature of 4N-IFN, also demonstrating a higher solubility of this variant as denoted by the absence of precipitation at the end of the experiment, in contrast with the NG-IFN behaviour. Furthermore, far-UV circular dichroism (CD) spectrum of 4N-IFN was virtually superimposed with that of NG-IFN, indicating that the IFN structure was not altered by the addition of carbohydrate moieties. The same conclusion could be inferred from limited proteolysis studies. Our results suggest that glycoengineering could be a useful strategy for protecting rhIFN-alpha2b from inactivation by various external factors and for overcoming aggregation problems during the production process and storage.

US$19.90

PMID: 20403411

DOI: 10.1016/j.biochi.2010.04.004


Other references

Pedrazzoli, S.; Pasquali, C.; Sperti, C.; Filipponi, C., 1999: Epidemiology of pancreatic cancer in northeast Italy Incidence, resectability rate, hospital stay and survival. Gastroenterology 116(4 PART 2): A1340, April

Goodin, M.S.; Thor, E.J.; Haworth, W.S., 1994: Use of computational fluid dynamics in the design of the Avecor Affinity oxygenator. Perfusion 9(3): 217-222

Schenk, P.; Stiebellehner, L.; Burghuber, O., C.; Kneussl, M.; Lang, I., M., 2005: Untersuchung des lungenkreislaufes mittels rechtsherzkatheter - Positionspapier des kardiopulmonalen arbeitskreises der Osterreichischen gesellschaft fur pneumologie und der Osterreichischen gesellschaft fur kardiologie. Wiener Klinische Wochenschrift 117(18): 651-662

Liu, T.Zhi.; Zhang, D.Sheng.; Liang, Y.; Zhou, N.Ning.; Gao, H.Fei.; Liu, K.Jun.; Wu, H.Ying., 2011: Treatment strategies and prognostic factors of patients with primary germ cell tumors in the mediastinum. The aim of this study was to evaluate the clinical characteristics and survival outcomes of patients with primary mediastinal germ cell tumor (PMGCT) by identifying the prognostic factors and efficacies of different treatment modalities. Fifty-fiv...

Saeki, Y., 1963: Effect of freezing point depression of diluent on the fertilising ability of cock sperm.. The mean freezing point depression (F.P.D.) of whole semen in winter and spring was 0.81 degrees C. Semen diluted with egg-citrate diluents with F.P.D.s of 0.615 degrees , 0.715 degrees and 0.925 degrees C gave similar fertility results to whole s...

Atanassov, N., 1972: Hymenopteraarten im westlichen 'Stara Planina' - Gebirge. Izvestiya na Zoologicheskiya lnstitut Sofiya, 36: 23-59

Dissado; Sweeney, 1993: Physical model for breakdown structures in solid dielectrics. Physical Review. B, Condensed Matter 48(22): 16261-16268

Kosec, G.; Alvarez, V.; Cazzulo, J., J., 2006: Cysteine proteinases of Trypanosoma cruzi: from digestive enzymes to programmed cell death mediators. Trypanosoma cruzi, the parasite causing Chagas disease, contains a number of proteolytic enzymes. The recent completion of the genome sequence of the T cruzi CL Brener clone suggests the presence of 70 cysteine peptidases, 40 serine peptidases (no...

Mihoci, I.; Delic, A.; Gjurasin, B.; Bucar, M.; Kucinic, M., 2007: First finding of the critically endangered butterfly Maculinea alcon (Denis & Schiffermuller, 1775) (Lepidoptera : Lycaenidae) in the Pannonian part of Croatia. The critically endangered butterfly Maculinea alcon was found at the locality Grubisnopoljska Bilogora in August 2006. That is the first finding of this species in the Pannonian part of Croatia and the second in Croatia as a whole. The biology of...

Chien, D.S.; Sandri, R.B.; Tang-Liu, D.S., 1992: Systemic pharmacokinetics of acitretin, etretinate, isotretinoin, and acetylenic retinoids in guinea pigs and obese rats. Etretinate, a highly lipophilic retinoid, is known to accumulate in the human body with a slow systemic elimination (half-life approximately 100 days) after long-term treatment. Retinoids with high lipophilicity and slow body elimination have the...