Application of the Protein Maker as a platform purification system for therapeutic antibody research and development

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• 作者：Geneviè ; ve Hé ; lie^a ; Marie Parat^a ; Fré ; dé ; ric Massé ; ^b ; Cory J. Gerdts^c ; Thomas P. Loisel^a ; Allan Matte^a ; ^{allan.matte@nrc-cnrc.gc.ca}
• 关键词：CHO ; Chinese Hamster Ovary ; DPBS ; Dulbecco's phosphate buffered saline ; HC ; IgG heavy chain ; HCP ; host cell protein ; IMAC ; immobilized metal ion affinity chromatography ; LC ; IgG light chain ; mAb ; monoclonal antibody ; OAA ; one-armed antibody
• 刊名：Computational and Structural Biotechnology Journal
• 出版年：2016
• 出版时间：2016
• 年：2016
• 卷：14
• 期：Complete
• 页码：238-244
• 全文大小：668 K
• 卷排序：14

文摘

Within the research and development environment, higher throughput, parallelized protein purification is required for numerous activities, from small scale purification of monoclonal antibodies (mAbs) and antibody fragments for in vitro and in vivo assays to process development and optimization for manufacturing. Here, we describe specific applications and associated workflows of the Protein Maker liquid handling system utilized in both of these contexts. To meet the requirements for various in vitro assays, for the identification and validation of new therapeutic targets, small quantities of large numbers of purified antibodies or antibody fragments are often required. Reducing host cell proteins (HCP) levels following capture with Protein A by evaluating various wash buffers is an example of how parallelized protein purification can be leveraged to improve a process development outcome. Stability testing under various conditions of in-process intermediates, as an example, the mAb product from a clarified harvest, requires parallelized protein purification to generate concurrent samples for downstream assays. We have found that the Protein Maker can be successfully utilized for small-to-mid scale platform purification or for process development applications to generate the necessary purified protein samples. The ability to purify and buffer exchange up to 24 samples in parallel offers a significant reduction in time and cost per sample compared to serial purification using a traditional FPLC system. By combining the Protein Maker purification system with a TECAN Freedom EVO liquid handler for automated buffer exchange we have created a new, integrated platform for a variety of protein purification and process development applications.