Development of a routine analysis method for liposome encapsulated recombinant interleukin-2

被引:10
作者
Koppenhagen, FJ
Storm, G
Underberg, WJM [2 ]
机构
[1] Univ Utrecht, Utrecht Inst Pharmaceut Sci, Dept Pharmaceut, NL-3508 TB Utrecht, Netherlands
[2] Univ Utrecht, Utrecht Inst Pharmaceut Sci, Dept Pharmaceut Anal, Utrecht, Netherlands
来源
JOURNAL OF CHROMATOGRAPHY B | 1998年 / 716卷 / 1-2期
关键词
recombinant interleukin-2; interleukin-2;
D O I
10.1016/S0378-4347(98)00271-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This paper describes the development of an isocratic reversed-phase high-performance Liquid chromatographic method for the routine analysis of recombinant interleukin-2 (rIL-2) in Liposome samples. The chromatographic system employed a C-4 column maintained at 30 degrees C eluted with 52.5% (w/w) acetonitrile in water, containing 100 mM NaClO4 and 10 mM HClO4. To remove phospholipid interference the chromatographic method was combined with a lipid-extraction procedure. No significant loss of rIL-2 was noted upon inclusion of this extraction step. The protein eluted from the column with a capacity factor (k') of 5.8. The method was validated for robustness, linearity, precision and reproducibility. It was shown that the method was linear over a sample concentration range of 1-100 mu g/ml. Upon assessment of the intra-day and inter-day precision, the relative standard deviations (RSD) were within the range of the methodical error (approximately 5%), except at the lower concentration of 10 mu g/ml, where the intra-day RSD was relatively high (17.8%). The recovery of rIL-2 upon liposome preparation and subsequent analysis of the samples was in the range 94+/-9%. The results indicate that the method is suitable for routine quantitation of rIL-2 in liposomal samples. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:285 / 291
页数:7
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