Heat-induced aggregation of recombinant erythropoietin in the intact and deglycosylated states as monitored by gel permeation chromatography combined with a low-angle laser light scattering technique.

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 Biochem

journal_title

Journal of biochemistry

authors

Endo Y,Nagai H,Watanabe Y,Ochi K,Takagi T

doi

10.1093/oxfordjournals.jbchem.a123961

keywords:

subject

Has Abstract

pub_date

1992-11-01 00:00:00

pages

700-6

issue

5

eissn

0021-924X

issn

1756-2651

journal_volume

112

pub_type

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