Integration of a complete downstream process for the automated lab-scale production of a recombinant protein
(2019) In Journal of Biotechnology 301. p.45-51- Abstract
In this work, an automated downstream process for the purification and formulation of a recombinant protein was integrated at lab scale in a single chromatography unit. The purification chain consists of three bind-and-elute chromatography columns, a flow-through membrane chromatography step, and a final ultrafiltration-diafiltration (UFDF) step to concentrate and formulate the sample. An integrated downstream process increases productivity and decreases process time and hold-up volume. In addition, the automation of the process allows reducing the manual work and increases reproducibility. To integrate the downstream steps, all the intermediate tanks are removed, and the eluate of a column is loaded directly onto the next one. This... (More)
In this work, an automated downstream process for the purification and formulation of a recombinant protein was integrated at lab scale in a single chromatography unit. The purification chain consists of three bind-and-elute chromatography columns, a flow-through membrane chromatography step, and a final ultrafiltration-diafiltration (UFDF) step to concentrate and formulate the sample. An integrated downstream process increases productivity and decreases process time and hold-up volume. In addition, the automation of the process allows reducing the manual work and increases reproducibility. To integrate the downstream steps, all the intermediate tanks are removed, and the eluate of a column is loaded directly onto the next one. This makes it necessary to design the process in order to minimize the column volumes and the process time. A research software called Orbit was used to automate the purification process and implement a UFDF step in the chromatography unit. The whole downstream sequence was successfully implemented at lab scale, getting a pure concentrated and formulated product with a productivity of 1.09 mg mL−1 h−1, achieving a time reduction from almost two to one working day, while getting a similar yield and purity. Regarding the UFDF operation, the sample was concentrated 10 times, and 97% of the old buffer was exchanged by the formulation buffer with a sequential diafiltration.
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- author
- Gomis-Fons, Joaquín LU ; Löfgren, Anton LU ; Andersson, Niklas LU ; Nilsson, Bernt LU ; Berghard, Lotta and Wood, Susanne
- organization
- publishing date
- 2019
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Automation, Chromatography, Downstream process, Integrated bioprocessing, Integrated column sequences, Ultrafiltration
- in
- Journal of Biotechnology
- volume
- 301
- pages
- 7 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:85067170119
- pmid:31145936
- ISSN
- 0168-1656
- DOI
- 10.1016/j.jbiotec.2019.05.013
- language
- English
- LU publication?
- yes
- id
- 3bb62f5e-894f-4e50-af50-cb0230b7bbbd
- date added to LUP
- 2019-06-26 12:37:11
- date last changed
- 2024-06-12 21:46:50
@article{3bb62f5e-894f-4e50-af50-cb0230b7bbbd, abstract = {{<p>In this work, an automated downstream process for the purification and formulation of a recombinant protein was integrated at lab scale in a single chromatography unit. The purification chain consists of three bind-and-elute chromatography columns, a flow-through membrane chromatography step, and a final ultrafiltration-diafiltration (UFDF) step to concentrate and formulate the sample. An integrated downstream process increases productivity and decreases process time and hold-up volume. In addition, the automation of the process allows reducing the manual work and increases reproducibility. To integrate the downstream steps, all the intermediate tanks are removed, and the eluate of a column is loaded directly onto the next one. This makes it necessary to design the process in order to minimize the column volumes and the process time. A research software called Orbit was used to automate the purification process and implement a UFDF step in the chromatography unit. The whole downstream sequence was successfully implemented at lab scale, getting a pure concentrated and formulated product with a productivity of 1.09 mg mL<sup>−1</sup> h<sup>−1</sup>, achieving a time reduction from almost two to one working day, while getting a similar yield and purity. Regarding the UFDF operation, the sample was concentrated 10 times, and 97% of the old buffer was exchanged by the formulation buffer with a sequential diafiltration.</p>}}, author = {{Gomis-Fons, Joaquín and Löfgren, Anton and Andersson, Niklas and Nilsson, Bernt and Berghard, Lotta and Wood, Susanne}}, issn = {{0168-1656}}, keywords = {{Automation; Chromatography; Downstream process; Integrated bioprocessing; Integrated column sequences; Ultrafiltration}}, language = {{eng}}, pages = {{45--51}}, publisher = {{Elsevier}}, series = {{Journal of Biotechnology}}, title = {{Integration of a complete downstream process for the automated lab-scale production of a recombinant protein}}, url = {{http://dx.doi.org/10.1016/j.jbiotec.2019.05.013}}, doi = {{10.1016/j.jbiotec.2019.05.013}}, volume = {{301}}, year = {{2019}}, }