Process Development and Scale-up for Vibrio natriegens: From BioLector XTMicrobioreactor to Pre-pilot Scale

Authors:
Tiphaine Clement, TWB
Joanna Abboud, TWB
Anna Kress & Noud Drummen, Beckman Coulter Life Sciences

Abstract

Optimization, scalability and transferability represent key challenges in bioprocess development. In
this study, scalable fermentation processes for Vibrio natriegens were explored using the BioLector XT
Microbioreactor. Initial cultivation parameters, including glycerol concentration (20, 30 and 50 g/L),
inoculation density (OD₆₀₀ 0.1-0.5) and induction timing were systematically evaluated. Different feeding
strategies (constant, linear, exponential) and feed medium compositions (1× and 4× VN medium) were
compared to optimize mScarlet production. Two optimized process conditions were subsequently
transferred to 20-L bioreactors to validate scalability. The high-performance process (4× VN feed,
feed start at four hours) achieved 1.8-fold higher biomass compared to the low-performance process
(1× VN feed, feed start at eight hours) in both scales. A strong correlation was observed between the
BioLector XT Microbioreactor and the 20-L bioreactor performance, demonstrating successful process
transferability. The data obtained in the BioLector XT instrument, regarding medium composition and
process strategy, was representative of what was observed in the 20-L bioreactor, allowing strong
confidence in the scalability and transferability of processes developed at microscale and providing
better understanding of V. natriegens physiology.