Application of the Protein Maker as a platform purification system for therapeutic antibody research and development.
Comput Struct Biotechnol J
; 14: 238-44, 2016.
Article
en En
| MEDLINE
| ID: mdl-27418955
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.
Antibody; CHO, Chinese Hamster Ovary; DPBS, Dulbecco's phosphate buffered saline; HC, IgG heavy chain; HCP, host cell protein; Hybridoma; IMAC, immobilized metal ion affinity chromatography; LC, IgG light chain; OAA, one-armed antibody; Parallelized protein purification; Process development; Protein A; Protein G; mAb, monoclonal antibody
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1
Colección:
01-internacional
Base de datos:
MEDLINE
Idioma:
En
Revista:
Comput Struct Biotechnol J
Año:
2016
Tipo del documento:
Article
País de afiliación:
Canadá
Pais de publicación:
Países Bajos