Abstract
Methods of biomass monitoring have increasingly been developed towards online, in situ techniques in order to advance process analysis and control. Off-line, ex situ methods, such as dry cell mass determination and direct cell counts, remain the reference for determining cell mass and number, respectively, but this type of analysis is time consuming. Absorbance measurement, which is used extensively as an off-line, ex situ, or online, in situ technique, is fast and straightforward, as the absorbance can be correlated to cell mass and number by a simple calibration. The downside is that absorbance measurements provide no estimation of viability and in situ applications can suffer from interference, such as aeration. Impedance spectroscopy is widely available and provides a quick measure of viable cell concentration, but does not give an estimation of total cell concentration and, hence, potential product. Sensitivity of impedance spectroscopy remains an issue at low cell concentration, and interference during in situ measurements is also a significant factor. In this chapter, a range of protocols is presented for online determination of biomass yields of recombinant yeast cultures.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Olsson L, Nielsen J (1997) On-line and in situ monitoring of biomass in submerged cultivations. Trends Biotechnol 15:517–522
Mallette MF (1969) Evaluation of growth by physical and chemical means. In: Norris JR, Ribbons DW (eds) Methods in microbiology. Academic, London, England and New York, NY, USA, pp 521–566
Junker BH, Reddy J, Gbewonyo K, Greasham R (1994) Online and in situ monitoring technology for cell-density measurement in microbial and animal-cell cultures. Bioprocess Eng 10:195–207
Singh A, Kuhad RC, Sahai V, Ghosh P (1994) Evaluation of biomass. Adv Biochem Eng Biotechnol 51:47–70
Clarke DJ, Blakecoleman BC, Carr RJG, Calder MR, Atkinson T (1986) Monitoring reactor biomass. Trends Biotechnol 4:173–178
Fehrenbach R, Comberbach M, Petre JO (1992) Online biomass monitoring by capacitance measurement. J Biotechnol 23:303–314
Kiviharju K, Salonen K, Moilanen U, Eerikainen T (2008) Biomass measurement online: the performance of in situ measurements and software sensors. J Ind Microbiol Biotechnol 35:657–65
Carvell JP, Dowd JE (2006) On-line measurements and control of viable cell density in cell culture manufacturing processes using radio-frequency impedance. Cytotechnol 50:35–48
Markx GH, Davey CL (1999) The dielectric properties of biological cells at radiofrequencies: applications in biotechnology. Enzyme Microbial Technol 25:161–171
Yardley YE, Kell DB, Barrett J, Davey CL (2000) On-line, real-time measurements of cellular biomass using dielectric spectroscopy. Biotechnol Genetic Eng Rev 17:3–35
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+business Media, LLC
About this protocol
Cite this protocol
Palmer, S.M., Kunji, E.R.S. (2012). Online Monitoring of Biomass Accumulation in Recombinant Yeast Cultures. In: Bill, R. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 866. Humana Press. https://doi.org/10.1007/978-1-61779-770-5_14
Download citation
DOI: https://doi.org/10.1007/978-1-61779-770-5_14
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-769-9
Online ISBN: 978-1-61779-770-5
eBook Packages: Springer Protocols