The Application of Dielectric Spectroscopy and Biocalorimetry for the Monitoring of Biomass in Immobilized Mammalian Cell Cultures

Abstract

Abstract:The purpose of this study was to introduce dielectric spectroscopy and biocalorimetry as monitoring methods to follow immobilised Chinese Hamster Ovary (CHO) cell culture development. The theory behind both monitoring techniques is explained and perfusion cultures are performed in a Reaction Calorimeter (eRC1 from Mettler Toledo) as an application example. The findings of this work show that dielectric spectroscopy gives highly reliable information upon the viable cell density throughout the entire culture. On the other hand, the RC1 could only provide accurate data from day 5, when the cell density exceeded 4 × 10 , a viable cell density commonly achieved in fed-batch and the early stages of a perfusion culture. This work suggests that biocalorimetry should be possible to implement at industrial scale to monitor CHO cell cultures.

Cytoplasmic conductivity as a marker for bioprocess monitoring: Study of Chinese hamster ovary cells under nutrient deprivation and reintroduction

“…Near‐infrared spectroscopy has been demonstrated to be useful in monitoring metabolites, viable cell density, and total cell density in batch cultures for final product quality control (Crowley, Arnold, Wood, Harvey, & McNeil, ; Teixeira, Oliveira, Alves, & Carrondo, ). Dielectric spectroscopy has been employed to monitor the biomass in batch cultures to improve process monitoring (Ansorge, Esteban, & Schmid, ; Cole, Demont, & Marison, ; Justice et al, ; Opel, Li, & Amanullah, ). One important aspect of mammalian cell cultures for which PAT tools have been developed is the metabolically active biomass (Ansorge et al, ; Handlogten et al, ; Zhang et al, ).…”

“…The use of dielectric properties for monitoring biomass is of interest for biopharmaceutical production as the methods are label‐free and noninvasive and have the potential to perform online analysis (Carvell & Dowd, ; Downey, Graham, Breit, & Glutting, ; Kiviharju et al, ; Lee, Carvell, Brorson, & Yoon, ; Petiot, Ansorge, Rosa‐Calatrava, & Kamen, ). It has been demonstrated that dielectric spectroscopy is a useful technique to monitor the biomass as well as the membrane and cytoplasm properties of Chinese hamster ovary (CHO) cells (Ansorge, Esteban, & Schmid, ; Ansorge et al, ; Cole et al, ; Downey et al, ; Elias, Zeiser, Bédard, & Kamen, ; Justice et al, ; Opel et al, ). Table shows the application of dielectric spectroscopy in monitoring dielectric parameters, specially cytoplasm conductivity in different cell lines.…”

“…The majority of the available dielectric based label‐free methods for monitoring biomass in a batch culture are based on bulk measurements. Dielectric spectroscopy techniques use the average dielectric properties of the cell culture medium to estimate the concentration of cells and their dielectric properties (Ansorge et al, ; Asami, Takahashi, & Takashima, ; Asami & Yamaguchi, ; Cannizzaro et al, ; Cole et al, ; Courtès et al, ; Druzinec et al, ; Grein et al, ; Justice et al, ; Katsumoto et al, ; Opel et al, ; Ron et al, ). Employing dielectric spectroscopy, also known as capacitance measurement, for real‐time in situ monitoring of the biomass content in fed‐batch cultures has been demonstrated to be successful in the estimation of dielectric properties and hence the physiological state of the cells (Ansorge et al, ; Petiot et al, ).…”

Control of IgG glycosylation by in situ and real‐time estimation of specific growth rate of CHO cells cultured in bioreactor

“…The need for biopharmaceutical protein products, in particular recombinant monoclonal antibodies (mAbs), has been constantly increasing in the last decade (Walsh, ). Chinese hamster ovary (CHO) cells are the most commonly used cell lines for mAb production due to their capability of performing human‐like posttranslational modifications (PTMs; Cole, Demont, & Marison, ). Glycosylation is one of the most important PTMs for mAbs, as it impacts effector functions, pharmacokinetics, antigenicity, safety, stability, and solubility of the mAbs produced (Tharmalingam, Wu, Callahan, & T. Goudar, ).…”

Zeta potential change of Neuro-2a tumor cells after exposure to alumina nanoparticles

“…Many concerns arise as regards to their potential toxicity to mammalian and also human cells. The following methods for studying cells characteristics were used: Raman spectroscopy [1], dielectric spectroscopy [2], and NMR spectroscopy techniques [3]. But these methods have a range of disadvantages such as relatively expensive reagents, heating and labour input.…”