کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020814 1069212 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Purification process development of a recombinant monoclonal antibody expressed in glycoengineered Pichia pastoris
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Purification process development of a recombinant monoclonal antibody expressed in glycoengineered Pichia pastoris
چکیده انگلیسی

A robust and scalable purification process was developed to quickly generate antibody of high purity and sufficient quantity from glycoengineered Pichia pastoris fermentation. Protein A affinity chromatography was used to capture the antibody from fermentation supernatant. A pH gradient elution was applied to the Protein A column to prevent antibody precipitation at low pH. Antibody from Protein A chromatography contained some product related impurities, which were the misassembling of cleaved heavy chain, heavy chain and light chain. It also had some process related impurities, including Protein A residues, endotoxin, host cell DNA and proteins. Cation exchange chromatography with optimal NaCl gradient at pH 4.5–6.0 efficiently removed these product and process related impurities. The antibody from glycoengineered P. pastoris was comparable to its commercial counterpart in heterotetramer folding, physical stability and binding affinity.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 76, Issue 1, March 2011, Pages 7–14
نویسندگان
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