|کد مقاله||کد نشریه||سال انتشار||مقاله انگلیسی||ترجمه فارسی||نسخه تمام متن|
|6452279||1417009||2016||13 صفحه PDF||سفارش دهید||دانلود رایگان|
- Sodium citrate precipitation was developed as the primary step for IgG purification.
- Chromatin-directed cell culture clarification elevated the purification capability.
- Surfactant effectively reduced the residual free antibody light chain during precipitation.
- Void-exclusion anion exchange chromatography was integrated with sodium citrate precipitation.
- A simple and efficient non-protein A based purification platform with only one-column step was established.
Protein A affinity chromatography, featured by its robustness and high-specificity, is still dominant as a first capture step for the purification of immunoglobulin G monoclonal antibodies (IgG mAbs). However, the material and operational costs of protein A are universally recognized as high, and its productivity is also limited as column mode. In order to overcome these limitations, industry is increasingly considering the use of non-protein A-based processes for IgG purification. In this study, sodium citrate precipitation (SCP) was developed as the primary purification step, and chromatin-directed cell culture clarification was demonstrated to significantly elevate the purification capability. Additional 0.05% (w/v) of Tween 20 was shown to effectively reduce the residual free antibody light chain (LC) during precipitation. The resuspended IgG was further polished by void-exclusion anion exchange chromatography (VEAX), which supported protein loading without buffer adjustment. The non-histone host cell protein (nh-HCP) content in the final product was about 5Â ppm and histone HCP below limit of detection (LOD). DNA was reduced to less than 1Â ppb, and aggregates/free LC less than 0.1%. The overall IgG recovery was 87.2%. A simple and efficient purification platform with only one-column step was therefore established, providing a more promising alternative to the current prevailing protein A-based purification platforms.
Journal: Journal of Biotechnology - Volume 236, 20 October 2016, Pages 128-140