EFFECT OF BUFFER CONSTITUENTS ON THE DETERMINATION OF THERAPEUTIC PROTEINS BY CAPILLARY ELECTROPHORESIS

被引:41
作者
GUZMAN, NA [1 ]
MOSCHERA, J [1 ]
IQBAL, K [1 ]
MALICK, AW [1 ]
机构
[1] HOFFMANN LA ROCHE INC, PHARMACEUT RES & DEV, NUTLEY, NJ 07110 USA
来源
JOURNAL OF CHROMATOGRAPHY | 1992年 / 608卷 / 1-2期
关键词
D O I
10.1016/0021-9673(92)87124-Q
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Capillary electrophoresis has proved to be a versatile method for the determination of proteins, peptides and amino acids in pharmaceutical formulations. For quantification of the capillary electrophoresis data, however, significant errors may result if the analysis is performed using improper separation conditions. The peak area response for protein analytes, which is generally low in conventional UV detection, may also vary dramatically depending on the nature of the buffer used in the separation. This paper describes the effects of various buffer constituents and analytical conditions on the capillary electrophoretic separation and quantification of a humanized monoclonal antibody in bulk form and in a typical therapeutic formulation. For optimum peak area response and reproducibility, protein derivatization with an appropriate chromophore (e.g., fluorescamine) and separation in the presence of a moderate ionic strength buffer containing lithium chloride, tetramethylammonium chloride or trimethylammonium propylsulfonate is recommended. General guidelines for the determination of proteins by capillary electrophoresis and a rationale for the use of internal standards to improve the quantification of data are also discussed.
引用
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页码:197 / 204
页数:8
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