Acta Pharm. 58 (2008) 199-206
Short communication
Studying the formation of aggregates in
recombinant human granulocyte-
-colony stimulating factor (rHuG-CSF), lenograstim, using size-exclusion
chromatography and SDS-PAGE
JASMINA TONIC RIBARSKA, SUZANA TRAJKOVIC JOLEVSKA, ANA POCEVA PANOVSKA and
ANETA DIMITROVSKA
jasminatonic@yahoo.com
Faculty of Pharmacy, University “Ss Cyril and
Methodius”, 1000
Accepted February 14, 2008
The stability of proteins is a subject of intense current interest.
Aggregation, as a dominant degradation pathway for therapeutic proteins, may
cause multiple adverse effects, including loss of efficacy and immunogenicity.
In the present study, the formation of aggregates in lenograstim under
physiological conditions was monitored. For this purpose, a simple and
selective size-exclusion high-performance liquid chromatography method for
detection and separation of aggregates from intact protein was developed.
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis was performed under
reducing and non-reducing conditions to determine the nature of aggregate bond
formation. Using both techniques, the presence of a low aggregate content
attached via disulfide bonds was detected.
Keywords: lenograstim, aggregate formation, size-exclusion
high-performance liquid chromatography, sodium dodecyl sulphate-polyacrylamide
gel electrophoresis