Studying the formation of aggregates in recombinant human granulocyte-colony stimulating factor (rHuG-CSF), lenograstim, using size-exclusion chromatography and SDS-PAGE.
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.
Faculty of Pharmacy, University Ss Cyril and Methodius, 1000 Skopje, Macedonia. jasminatonic@yahoo.com
Jasmina Tonic
JT
Ribarska
Suzana Trajkovic
ST
Jolevska
Ana Poceva
AP
Panovska
Aneta
A
Dimitrovska
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry*; D006801:Humans; D000078224:Lenograstim; D008970:Molecular Weight; D010316:Particle Size; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry
D004220:Disulfides; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D000078224:Lenograstim