Abstract:
:Aggregate formation of recombinant human erythropoietin (r-EPO) on heat-treatment was followed by gel permeation chromatography combined with a low-angle laser light scattering technique under various conditions with respect to pH and salt concentration in order to provide basic knowledge about the change strictly required to be monitored for medicinal proteins. When heated at 60 degrees C at neutral pH, an aggregate with a limited size consisting of about 20 r-EPO molecules was formed. On heating at 50 degrees C at acidic pH, aggregation was unlimited. The aggregation proceeded non-covalently in acidic pH, but in alkaline pH covalent bond formation was also involved. Increase in salt concentration enhanced the aggregation. Deglycosylation of the N-linked oligosaccharides made r-EPO remarkably susceptible to aggregation on heat-treatment, indicating that the carbohydrate chains are essential to the stability of r-EPO.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Endo Y,Nagai H,Watanabe Y,Ochi K,Takagi Tdoi
10.1093/oxfordjournals.jbchem.a123961keywords:
subject
Has Abstractpub_date
1992-11-01 00:00:00pages
700-6issue
5eissn
0021-924Xissn
1756-2651journal_volume
112pub_type
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