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Integration of a complete downstream process for the automated lab-scale production of a recombinant protein

Gomis-Fons, Joaquín LU ; Löfgren, Anton LU orcid ; Andersson, Niklas LU orcid ; Nilsson, Bernt LU ; Berghard, Lotta and Wood, Susanne (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
; ; ; ; and
organization
publishing date
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
  • pmid:31145936
  • scopus:85067170119
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
2024-03-03 17:15:12
@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}},
}