Development and calibration of a standard for the protein content of granulocyte colony-stimulating factor products

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• 作者：Kai Gao ; Chunming Rao¹ ; Lei Tao ; Chunmei Han ; Xinchang Shi ; Lan Wang ; Wenhong Fan ; Lei Yu ; Junzhi Wang ; ^{wangjz@nicpbp.org.cn}
• 关键词：Granulocyte colony-stimulating factor (G-CSF) ; Protein content ; Standard ; Kjeldahl method ; High performance liquid chromatography (HPLC)
• 刊名：Biologicals
• 出版年：2012
• 期刊代码：162_10451056
• 类别：bio
• 出版时间：March, 2012
• 卷：40
• 期：2
• 页码：151-157
• 文件大小：153 K

摘要

This collaborative study characterizes a homogeneous standard for the protein content determination of granulocyte colony-stimulating factor (G-CSF) products with traceability of the measurement. The Kjeldahl method was used to determine the average protein content of G-CSF bulk as 2.505聽mg/ml (95%C.I: 2.467-2.543聽mg/ml, GCV 4.0%). Using G-CSF bulk as a traceability benchmark, the protein content of the final freeze-dried standard using reverse phase HPLC (RP-HPLC) was 215.4聽渭g protein per ampoule (95%C.I: 212.407-218.486聽渭g/ampoule, GCV 3.4%). A comparative study showed that there was no difference between using Filgrastim CRS (European Pharmacopeia G-CSF reference standard) and freeze-dried homogeneous standard when quantifying G-CSF protein content by RP-HPLC (P聽>聽0.05). However, there were significant differences in the G-CSF protein content obtained using a serum albumin standard by Lowry assay and a G-CSF standard with RP-HPLC. Therefore, use of RP-HPLC with a freeze-dried homogeneous standard would eliminate the systematic errors introduced when using a serum albumin standard because of the differences in protein composition between the standard and the sample. It would also be helpful to use this method to compare the quality of G-CSF biosimilar products in situations where the protein content has been calibrated using various standards.