Quality Control and Stability Studies with the Monoclonal Antibody, Trastuzumab: Application of 1D-vs. 2D-Gel Electrophoresis

被引:26
作者
Nebija, Dashnor [1 ]
Noe, Christian R. [2 ]
Urban, Ernst [2 ]
Lachmann, Bodo [2 ]
机构
[1] Fac Med, Dept Pharmaceut Chem, Pristina 10000, Kosovo
[2] Univ Vienna, Dept Pharmaceut Chem, Fac Life Sci, Althanstr 14, A-1090 Vienna, Austria
关键词
trastuzumab; 2D-electophoresis; SDS-PAGE; charge heterogeneity; stability; 2-DIMENSIONAL GEL-ELECTROPHORESIS; CAPILLARY-ELECTROPHORESIS; MASS-SPECTROMETRY; PROTEINS; VALIDATION; MS; ELISA;
D O I
10.3390/ijms15046399
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Recombinant monoclonal antibodies (rmAbs) are medicinal products obtained by rDNA technology. Consequently, like other biopharmaceuticals, they require the extensive and rigorous characterization of the quality attributes, such as identity, structural integrity, purity and stability. The aim of this work was to study the suitability of gel electrophoresis for the assessment of charge heterogeneity, post-translational modifications and the stability of the therapeutic, recombinant monoclonal antibody, trastuzumab. One-dimensional, SDS-PAGE, under reducing and non-reducing conditions, and two-dimensional gel electrophoresis were used for the determination of molecular mass (Mr), the isoelectric point (pI), charge-related isoform patterns and the stability of trastuzumab, subjected to stressed degradation and long-term conditions. For the assessment of the influence of glycosylation in the charge heterogeneity pattern of trastuzumab, an enzymatic deglycosylation study has been performed using N-glycosidase F and sialidase, whereas carboxypeptidase B was used for the lysine truncation study. Experimental data documented that 1D and 2D gel electrophoresis represent fast and easy methods to evaluate the quality of biological medicinal products. Important stability parameters, such as the protein aggregation, can be assessed, as well.
引用
收藏
页码:6399 / 6411
页数:13
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