The impact of sialylation linkage-type on the pharmacokinetics of recombinant butyrylcholinesterases |
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Authors: | Cheng-Yu Chung Qiong Wang Shuang Yang Sandra Chough Younji Seo John F Cipollo Joseph P Balthasar Michael J Betenbaugh |
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Institution: | 1. Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland;2. Laboratory for Bacterial Polysaccharides, Division of Bacterial, Parasitic and Allergenic Products (DBPAP), Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, Maryland;3. Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, New York |
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Abstract: | Chinese hamster ovary (CHO) cells typically produce glycoproteins with N-glycans terminating in α-2,3 sialylation. Human cells produce glycoproteins that include α-2,3 and α-2,6 sialic acids. To examine the impact of altering protein sialylation on pharmacokinetic properties, recombinant human butyrylcholinesterase (BChE) was produced in CHO cells by knocking out the α-2,3 sialyltransferase genes followed by overexpression of the α-2,6 sialyltransferase (26BChE) enzyme. The N-glycan composition of 26BChE was compared to BChE with α-2,3 sialylation (23BChE) derived from wild-type CHO cells. Both 23BChE and 26BChE exhibited comparable antennarity distributions with bi-antennary di-sialylated glycans representing the most abundant glycoform. CD-1 mice were intravenously injected with the 23BChE or 26BChE, and residual BChE activities from blood collected at various time points for pharmacokinetic analyses. Although 23BChE contained a slightly lower initial sialylation level compared to 26BChE, the molecule exhibited higher residual activity between 5 and 24 hr postinjection. Pharmacokinetic analyses indicated that 23BChE exhibited an increase in area under the curve and a lower volume of distribution at steady state than that of 26BChE. These findings suggest that the type of sialylation linkage may play a significant role in the pharmacokinetic behavior of a biotherapeutic when tested in in vivo animal models. |
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Keywords: | butyrylcholinesterase pharmacokinetic sialic Acid α-2 3 sialylation α-2 6 sialylation |
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