Correlation of liquid chromatographic and biological assay for potency assessment of filgrastim and related impurities

Assessment of the Effects of Glycosylation on the Pattern and Kinetics of Degradation of Lenograstim in Comparison to Filgrastim Using a Stability-Indicating Orthogonal Testing Protocol

“…Susceptibility to oxidation has been linked with the methionine residues in rHuG-CSF and conversion of its sulfur-containing residues into sulfoxide resulted in loss of biological activity [16,29,30]. It should be noted that oxidized forms are not only possible degradation products but also product-related impurities that should be tested for in the final preparation [11,26].…”

“…It is also crucial for the development and validation of stability-indicating analysis protocols [14,15]. Such protocols are employed also during drug development stages to provide evidence of how the quality of a drug substance or drug product changes over time in response to a variety of environmental factors [15,16]. Since the introduction of FGM to the market, several reports have been published highlighting the extreme sensitivity of FGM to various physicochemical stress factors.…”

Analytical aspects of biosimilarity issues of protein drugs

“…In vivo and in vitro potency assays are more costly, time consuming and less robust than conventional analytical methods. In their pioneering work Skrlin et al tried to find an alternative liquid chromatography method that would replace in vitro proliferation of neutrophil granulocytes bioassay of filgrastim [42]. They established an RP-HPLC test for the measurement of impurities and degradants in filgrastim product.…”

“…Robust oxidation of filgrastim for example, decreased its original bioactivity down to 25%, while filgrastim dimer had 67% potency of that of intact filgrastim [42]. Three copy products of recombinant human Erythropoetin (Epo), manufactured in Korea, showed higher bioactivity compared to the originator product, and contained more glycoforms and impurities presumably due to different cell lines and manufacturing processes [43].…”

Purity assessment of recombinant human granulocyte colony‐stimulating factor in finished drug product by capillary zone electrophoresis

“…Various chromatographic techniques and methods provide information about presence of the contaminants in liquid filgrastim formulations, but none of them can be used as standalone to provide satisfactory information on the purity of the product. It has been shown that there is an inverse correlation between the content of related impurities and biological activity of filgrastim in injections using liquid chromatographic methods and mathematical model for the estimation of biological activity . CZE was used to achieve separation of one nonglycosylated and two glycosylated forms of filgrastim, while the estimation of the purity of nonglycosylated filgrastim was achieved using CIEF with characterization by MS .…”