Focussed beam reflectance measurement (FBRM) monitoring of particle size and morphology in suspension cultures ofMorinda citrifoliaandCentaurea calcitrapa

Abstract

Laser light scattering technology, as applied in the Lasentec focussed beam reflectance measurement (FBRM) system, was used to characterise two morphologically dissimilar plant cell suspension cultures, Morinda citrifolia and Centaurea calcitrapa. Shake-flask suspensions were analysed in terms of biomass concentration and aggregate size/shape over the course of typical batch growth cycles. For the heavily aggregated C. calcitrapa, biomass levels [from 10-160 g fresh weight (fw) l(-1))] were linearly correlated with FBRM counts. For M. citrifolia, which grows in unbranched chains of 2-10 elongated cells, linear correlation of biomass concentration with FBRM counts was applicable in the range 0-100 g fw l(-1); at higher levels (100-300 g fw l(-1)), biomass was non-linearly correlated with FBRM counts and length-weighted average FBRM chord length. For both cell systems, particle morphology (size/shape) was quantified using semi-automated digital image analysis. The average aggregate equivalent diameter (C. calcitrapa) and average chain length (M. citrifolia), determined using image analysis, closely tracked the FBRM average chord length. The data clearly demonstrate the potential for applying the FBRM technique for rapid characterisation of plant cell suspension cultures.

Use of focussed beam reflectance measurement (FBRM) for monitoring changes in biomass concentration

“…The methods used to grow and morphologically analyse cultures of M. citrifolia and C. calcitrapa have been presented previously [16]. In brief, batch cultures of M. citrifolia and C. calcitrapa were grown in shake flasks.…”

“…Jeffers et al [16] demonstrated that the biomass concentrations in plant cell cultures of both Centaurea calcitrapa (C. calcitrapa) and Morinda citrifolia (M. citrifolia) correlated well with total counts measured. C. calcitrapa grew as aggregates of 50-3,000 lm in size and cultures of 10-160 g FCW L -1 were linearly correlated.…”

A PAT Approach to Improve Process Understanding of High Shear Wet Granulation Through In-Line Particle Measurement Using FBRM C35

“…FBRM has been used to characterize a variety of particulate systems and its use in industry and academia is widespread, for example, crystallization, 13 polymerization, 14 wastewater treatment, 15 flocculation, 16 and plant cell suspensions. 17 The FBRM Model C35 system is equipped with a mechanical scraper, which can clear the optical window at a pre-defined time interval. This new feature allows the probe to be used in wet and sticky environments, without having to stop the process and clean the probe.…”

Optimization of Insect Cell Based Protein Production Processes - Online Monitoring, Expression Systems, Scale Up

“…Although the probe has already been utilized for offline characterization of microbial [95][96][97][98] and plant cell systems [99][100][101], applications in cell culture processes are not yet reported. The latest version of the G400 FBRM system distributed by Mettler-Toledo is fully autoclavable and therefore allows the integration of this technology even in cell culture processes.…”