کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2486134 | 1556514 | 2011 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A HighâThroughput UPLC Method for the Characterization of Chemical Modifications in Monoclonal Antibody Molecules
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کلمات کلیدی
موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
اکتشاف دارویی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Development of highâthroughput release and characterization assays is critical for the effective support of the rapidly growing biologics pipeline for biotherapeutics. Clipping of polypeptide chains is commonly monitored during process optimization, formulation development, and stability studies. A reduced capillary electrophoresisâsodium dodecyl sulfate (rCE -SDS) method is often used as a purity release assay for monitoring clips in monoclonal antibodies (mAbs); however, it has a cycle time of approximately 40 min, which is not suited for highâthroughput screening. Additionally, the characterization of clips and variants from electropherograms is not straightforward and takes significant time. Reduced reversedâphase (RP) chromatography has been a popular assay for the characterization and identification of clips and variants because it can be directly coupled with online mass spectrometric analysis. However, the highâcolumn temperature and low pH required for RP assays can induce onâcolumn cleavage and therefore skew the results. To minimize onâcolumn degradation, we have developed a highâthroughput method with a significantly shorter cycle time of 5 min. The short cycle time was achieved using an ultraâhighâpressure liquid chromatography (UPLC) system with a 1.7 μm phenyl column. This UPLC method allowed quantitation of hinge clipping in an IgG1 molecule and acid induced aspartic acid/proline (D/P) clip in an IgG2 molecule. The results from the UPLC method were comparable to those obtained with rCE-SDS. Additionally, the phenyl column offered partial resolution of oxidation and other chemical modifications, making this technique an attractive assay for highâthroughput process characterization and formulation screens.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical Sciences - Volume 100, Issue 12, December 2011, Pages 5115-5125
Journal: Journal of Pharmaceutical Sciences - Volume 100, Issue 12, December 2011, Pages 5115-5125
نویسندگان
Nicole Stackhouse, Amanda K. Miller, Himanshu S. Gadgil,