Synchronous fluorescence spectroscopy as a novel tool to enable PAT applications in bioprocesses.
Biotechnol Bioeng
; 108(8): 1852-61, 2011 Aug.
Article
em En
| MEDLINE
| ID: mdl-21391211
ABSTRACT
In this work, synchronous fluorescence spectroscopy (SFS) is evaluated as a new tool for real-time bioprocess monitoring of animal cell cultures. This technique presents several advantages over the traditional two-dimensional (2D) fluorometry since it provides data on various fluorescent compounds in a single spectrum, showing improved peak resolution and recording speed. Bioreactor cultures of three monoclonal antibody-producing CHO cell lines were followed in situ by both 2D and synchronous fluorometry techniques. The time profiles of the main spectral features in each data type present some differences, but principal component analysis indicated both as containing enough information to distinguish the cultures. Partial least squares regression models were then independently developed for viable cell density and antibody levels on the basis of the different fluorescence signals recorded, hiding half of the dataset for subsequent validation purposes. Regardless of the signal used, model predictions fit very well the off-line measurements; still, the synchronous spectra collected at a wavelength difference of 20 nm allowed comparable and superior performances for cell density and antibody titer, respectively, with validation accuracies higher than 91%. Therefore, SFS compares favorably with the traditional 2D approach, becoming an improved, faster option for real-time monitoring of cells and product titer over culture time. The readiness in data acquisition facilitates the design of process control strategies meeting the requirements of a PAT application.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Fluorescência
/
Biotecnologia
Tipo de estudo:
Evaluation_studies
/
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Biotechnol Bioeng
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Portugal