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Open AccessArticle

Estimating Extrinsic Dyes for Fluorometric Online Monitoring of Antibody Aggregation in CHO Fed-Batch Cultivations

Institute of Applied Biotechnology, Biberach University of Applied Science, 77781 Biberach, Germany
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Academic Editor: Christoph Herwig
Bioengineering 2017, 4(3), 65; https://doi.org/10.3390/bioengineering4030065
Received: 24 June 2017 / Revised: 15 July 2017 / Accepted: 17 July 2017 / Published: 24 July 2017
(This article belongs to the Special Issue Hybrid Modelling and Multi-Parametric Control of Bioprocesses)
Multi-wavelength fluorescence spectroscopy was evaluated in this work as tool for real-time monitoring of antibody aggregation in CHO fed-batch cultivations via partial least square (PLS) modeling. Therefore, we used the extrinsic fluorescence dyes 1-anilinonaphthalene-8-sulfonate (ANS), 4,4′-bis-1-anilinonaphthalene-8-sulfonate (Bis-ANS), or Thioflavin T (ThT) as medium additives. This is a new application area, since these dyes are commonly used for aggregate detection during formulation development. We determined the half maximum inhibitory concentrations of ANS (203 ± 11 µmol·L−1), Bis-ANS (5 ± 0.5 µmol·L−1), and ThT (3 ± 0.2 µmol·L−1), and selected suitable concentrations for this application. The results showed that the emission signals of non-covalent dye antibody aggregate interaction superimposed the fluorescence signals originating from feed medium and cell culture. The fluorescence datasets were subsequently used to build PLS models, and the dye-related elevated fluorescence signals dominated the model calibration. The soft sensors based on ANS and Bis-ANS signals showed high predictability with a low error of prediction (1.7 and 2.3 mg·mL−1 aggregates). In general, the combination of extrinsic dye and used concentration influenced the predictability. Furthermore, the ThT soft sensor indicated that the intrinsic fluorescence of the culture might be sufficient to predict antibody aggregation online. View Full-Text
Keywords: antibody aggregation; ANS; bioprocess monitoring; Bis-ANS; fluorescence spectroscopy; CHO antibody aggregation; ANS; bioprocess monitoring; Bis-ANS; fluorescence spectroscopy; CHO
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MDPI and ACS Style

Schwab, K.; Hesse, F. Estimating Extrinsic Dyes for Fluorometric Online Monitoring of Antibody Aggregation in CHO Fed-Batch Cultivations. Bioengineering 2017, 4, 65. https://doi.org/10.3390/bioengineering4030065

AMA Style

Schwab K, Hesse F. Estimating Extrinsic Dyes for Fluorometric Online Monitoring of Antibody Aggregation in CHO Fed-Batch Cultivations. Bioengineering. 2017; 4(3):65. https://doi.org/10.3390/bioengineering4030065

Chicago/Turabian Style

Schwab, Karen; Hesse, Friedemann. 2017. "Estimating Extrinsic Dyes for Fluorometric Online Monitoring of Antibody Aggregation in CHO Fed-Batch Cultivations" Bioengineering 4, no. 3: 65. https://doi.org/10.3390/bioengineering4030065

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