engleski

Protein self-association

Monomeric protein structure can form dimers, trimers and other aggregates induced by different types of stressors. During protein association process different types of bonding may occur, such as covalent, especially disulphide bonds, and non- covalent bonds: hydrogen bonds, electrostatic interactions, van der Waals interactions and hydrophobic bonds. Protein structure complexity makes mechanism of associates emergence unknown. Affected by these stressors, covalent and non- covalent bondage may occur and produce irreversible or reversible protein aggregates. Irreversible aggregates can be produced through heating, freezing-thawing, over-concentrating, isomerization, oxidation etc. In other hand, reversible aggregates are self-associates that are formed by agitation process. We examined formation of dimers, trimers and tetramers on rHuG-CSG, also known as granulocyte colony stimulating factor, induced by agitation through time period of: 60, 120, 180, 240, 300, 360, 420, 480, 540 and 600 s. Analysis was performed immediately after agitation by size exclusion chromatography, at pH= 7.0 (NH4HCO3 mobile phase). Due to increased pressure caused by centripetal acceleration during rotation monomeric structures merges and makes dimers firstly, then trimers, tetramers and other aggregates. By increasing agitation time the amount of reversible self-associates significantly increase.

HPLC ; rHu-G-CSF ; filgrastim ; agitation

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