کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1221574 1494661 2013 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Analysis and characterization of aggregation of a therapeutic Fc-fusion protein
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Analysis and characterization of aggregation of a therapeutic Fc-fusion protein
چکیده انگلیسی

Protein aggregation was observed in a purification intermediate of a therapeutic Fc-fusion protein stored at −30 °C, even though the protein was stable at 4 and −80 °C. The protein was expressed in Escherichia coli as an inclusion body, refolded, and purified using chromatography columns. To study the nature of this aggregation, a series of experiments were conducted to investigate factors that contributed to the protein instability during freezing. We found that the presence of free thiols in the protein is the intrinsic cause. The free thiol cross-linking sites were determined to be at the peptide moiety of the Fc-fusion protein using LC–MS. Partially frozen accompanied by the elevated pH and increased salt and protein concentrations were identified as extrinsic factors that facilitated the aggregation. These results provided important insights into purification process improvement and solution storage of this Fc-fusion protein.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical and Biomedical Analysis - Volume 72, 18 January 2013, Pages 59–64
نویسندگان
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