Premium
Integration of host strain bioengineering and bioprocess development using ultra‐scale down studies to select the optimum combination: An antibody fragment primary recovery case study
Author(s) -
Aucamp Jean P.,
Davies Richard,
Hallet Damien,
Weiss Amanda,
TitchenerHooker Nigel J.
Publication year - 2014
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25259
Subject(s) - bioprocess , fragment (logic) , strain (injury) , host (biology) , primary (astronomy) , scale (ratio) , computational biology , chemistry , biochemical engineering , biology , computer science , engineering , chemical engineering , genetics , physics , algorithm , quantum mechanics , astronomy , anatomy
An ultra scale‐down primary recovery sequence was established for a platform E. coli Fab production process. It was used to evaluate the process robustness of various bioengineered strains. Centrifugal discharge in the initial dewatering stage was determined to be the major cause of cell breakage. The ability of cells to resist breakage was dependant on a combination of factors including host strain, vector, and fermentation strategy. Periplasmic extraction studies were conducted in shake flasks and it was demonstrated that key performance parameters such as Fab titre and nucleic acid concentrations were mimicked. The shake flask system also captured particle aggregation effects seen in a large scale stirred vessel, reproducing the fine particle size distribution that impacts the final centrifugal clarification stage. The use of scale‐down primary recovery process sequences can be used to screen a larger number of engineered strains. This can lead to closer integration with and better feedback between strain development, fermentation development, and primary recovery studies. Biotechnol. Bioeng. 2014;111: 1971–1981. © 2014 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.