title: Suppression of IgM Proteolysis by Conformational Stabilization Through Excipients.
abstract: Protease activity from host cell lines may cause product loss or affect the quality of recombinant proteins. In this study, we showed that excipients like glycine and sorbitol reduce the proteolysis of an immunoglobulin M (IgM) in the presence of added proteases like α-chymotrypsin, papain, and pepsin. The activity of the proteases in the IgM-protective environments was conserved or even enhanced as tested using low molecular weight substrates. Thus, a higher resistance against proteolytic degradation appears to be caused by the conformational stabilization of the IgM due to preferential exclusion of sorbitol and glycine.
authors.affiliation: Department of Pharmaceutical Technology and Biopharmaceutics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria; Bioprocessing Technology Institute, ASTAR, 20 Biopolis Way, #06-01, Centros, 138668, Singapore.
authors.forename: Monika
authors.initials: M
authors.lastname: Mueller
authors.affiliation: Bioprocessing Technology Institute, ASTAR, 20 Biopolis Way, #06-01, Centros, 138668, Singapore.
authors.forename: Maybelle Q T
authors.initials: MQ
authors.lastname: Loh
authors.affiliation: Bioprocessing Technology Institute, ASTAR, 20 Biopolis Way, #06-01, Centros, 138668, Singapore.
authors.forename: Pete
authors.initials: P
authors.lastname: Gagnon
keywords: Conformational stability; Glycine; IgM; Proteolysis; Sorbitol
references.citation: Biotechnol Lett. 2003 Nov;25(22):1949-52
references.pmid: 14719832
references.citation: J Pharm Sci. 2008 Apr;97(4):1414-26
references.pmid: 17721938
references.citation: J Chromatogr A. 2009 Nov 6;1216(45):7851-64
references.pmid: 19815220
references.citation: J Biotechnol. 1991 Oct;20(3):249-61
references.pmid: 1367572
references.citation: In Vitro Cell Dev Biol Anim. 2005 Nov-Dec;41(10):330-6
references.pmid: 16448222
references.citation: J Biol Chem. 2001 Aug 3;276(31):28789-98
references.pmid: 11373282
references.citation: Stem Cells. 2008 Jun;26(6):1454-63
references.pmid: 18356574
references.citation: Biotechnol J. 2014 Apr;9(4):493-500
references.pmid: 24478053
references.citation: Protein Sci. 1997 Jan;6(1):211-21
references.pmid: 9007993
references.citation: J Pharm Sci. 1996 Apr;85(4):419-26
references.pmid: 8901081
references.citation: J Biol Chem. 2002 May 31;277(22):20079-86
references.pmid: 11896058
references.citation: J Biol Chem. 2001 May 25;276(21):18146-52
references.pmid: 11279138
references.citation: Haematologica. 2015 Mar;100(3):377-84
references.pmid: 25637055
references.citation: Appl Biochem Biotechnol. 2013 Feb;169(4):1431-48
references.pmid: 23315232
references.citation: Pharm Res. 2013 Mar;30(3):735-50
references.pmid: 23143394
references.citation: Biochim Biophys Acta. 2006 Jul;1764(7):1234-42
references.pmid: 16797260
references.citation: Biochem J. 2006 Sep 15;398(3):605-11
references.pmid: 16787388
references.citation: Biotechnol Bioeng. 2009 Dec 15;104(6):1132-41
references.pmid: 19655395
references.citation: Arch Biochem Biophys. 1983 Jul 1;224(1):169-77
references.pmid: 6870251