ON-LINE MONITORING OF BIOMASS CONCENTRATION BASED ON A CAPACITANCE SENSOR: ASSESSING THE METHODOLOGY FOR DIFFERENT BACTERIA AND YEAST HIGH CELL DENSITY FED-BATCH CULTURES

The performance of an in-situ capacitance sensor for on-line monitoring of biomass concentration was evaluated for some of the most important microorganisms in the biotechnology industry: Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Bacillus megaterium. A total of 33 batch and fed-batch cultures were carried out in a bench-scale bioreactor and biomass formation trends were followed by dielectric measurements during the growth phase as well as the induction phase, for 5 recombinant E. coli strains. Permittivity measurements and viable cellular concentrations presented a linear correlation for all the studied conditions. In addition, the permittivity signal was further used for inference of the cellular growth rate. The estimated specific growth rates mirrored the main trends of the metabolic states of the different cells and they can be further used for setting-up control strategies in fed-batch cultures.

[1]  Brandon J Downey,et al.  A Novel Approach for Using Dielectric Spectroscopy to Predict Viable Cell Volume (VCV) in Early Process Development , 2013, Biotechnology progress.

[2]  T. C. Zangirolami,et al.  Cloning, Auto-induction Expression, and Purification of rSpaA Swine Erysipelas Antigen , 2012, Current Microbiology.

[3]  A. M. Vélez,et al.  High-throughput strategies for penicillin G acylase production in rE. coli fed-batch cultivations , 2014, BMC Biotechnology.

[4]  H M Davey,et al.  Oscillatory, stochastic and chaotic growth rate fluctuations in permittistatically controlled yeast cultures. , 1996, Bio Systems.

[5]  R. Giordano,et al.  Non-conventional induction strategies for production of subunit swine erysipelas vaccine antigen in rE. coli fed-batch cultures , 2013, SpringerPlus.

[6]  Thomas Maskow,et al.  Observation of non-linear biomass-capacitance correlations: reasons and implications for bioprocess control. , 2008, Biosensors & bioelectronics.

[7]  N. Tripathi.  High Yield Production of Heterologous Proteins with Escherichia coli , 2009 .

[8]  A. Farnoud,et al.  Recent Advances in High Cell Density Cultivation for Production of Recombinant Protein , 2008 .

[9]  Ana P Ferreira,et al.  Evaluation of a new annular capacitance probe for biomass monitoring in industrial pilot-scale fermentations. , 2005, Journal of biotechnology.

[10]  J. García,et al.  Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Immobilization on Ionic Exchangers , 2006, Applied and Environmental Microbiology.

[11]  F. Studier,et al.  Protein production by auto-induction in high density shaking cultures. , 2005, Protein expression and purification.

[12]  Luis A. García,et al.  Application of flow cytometry to industrial microbial bioprocesses , 2010 .

[13]  V. M. Gonçalves,et al.  Development of production and purification processes of recombinant fragment of pneumococcal surface protein A in Escherichia coli using different carbon sources and chromatography sequences , 2011, Applied Microbiology and Biotechnology.

[14]  Tim W. Overton,et al.  Flow cytometric analysis of E. coli on agar plates: implications for recombinant protein production , 2014, Biotechnology Letters.

[15]  Michal Dabros,et al.  Simple control of specific growth rate in biotechnological fed-batch processes based on enhanced online measurements of biomass , 2010, Bioprocess and biosystems engineering.

[16]  Geoffrey Esteban,et al.  Impact of microcarrier coverage on using permittivity for on-line monitoring high adherent Vero cell densities in perfusion bioreactors , 2013 .

[17]  R. Giordano,et al.  Brazilian Journal of Chemical Engineering A SUPERVISION AND CONTROL TOOL BASED ON ARTIFICIAL INTELLIGENCE FOR HIGH CELL DENSITY CULTIVATIONS , 2014 .

[18]  L. Harvey,et al.  Heterologous protein production using the Pichia pastoris expression system , 2005, Yeast.

[19]  Joseph Shiloach,et al.  Growing E. coli to high cell density--a historical perspective on method development. , 2005, Biotechnology advances.

[20]  Ju Chu,et al.  Real-time viable-cell mass monitoring in high-cell-density fed-batch glutathione fermentation by Saccharomyces cerevisiae T65 in industrial complex medium. , 2008, Journal of bioscience and bioengineering.

[21]  K. Sathyaseelan.  High Yield Production of Heterologous Proteins with Escherichia coli , 2009 .

[22]  U. Rinas,et al.  Comparison of temperature- and isopropyl-β-d-thiogalacto-pyranoside-induced synthesis of basic fibroblast growth factor in high-cell-density cultures of recombinant Escherichia coli , 1995 .

[24]  C. Ghommidh,et al.  Dielectric monitoring of growth and sporulation of Bacillus thuringiensis , 2005, Biotechnology Letters.

[25]  Christopher L. Davey,et al.  The dielectric properties of biological cells at radiofrequencies : Applications in biotechnology , 1999 .

[26]  K. Kiviharju,et al.  Biomass measurement online: the performance of in situ measurements and software sensors , 2008, Journal of Industrial Microbiology & Biotechnology.

[27]  T. C. Zangirolami,et al.  Intensification of high cell-density cultivations of rE. coli for production of S. pneumoniae antigenic surface protein, PspA3, using model-based adaptive control , 2012, Bioprocess and Biosystems Engineering.