In-line monitoring of Bordetella pertussis cultivation using fluorescence spectroscopy

Fluorescence spectroscopy is a non-invasive and highly sensitive method for bioprocess monitoring. The use of fluorescence spectroscopy is not very well established in the industry for in-line monitoring. In the present work, a 2-D fluorometer with two excitation lights (365 and 405 nm) and emission spectra in the range of 350-850 nm were used for in-line monitoring of two strains of Bordetella pertussis cultivation operated in batch and fed batch. A Partial Least Squares (PLS) based regression model was used for the estimation of cell biomass, amino acids (glutamate and proline) and antigen (Pertactin) produced. It was observed that accurate predictions were achieved when models were calibrated separately for each cell strain and nutrient media formulation. Also, prediction accuracy was improved when dissolved oxygen, agitation and culture volume are added as additional features in the regression model. The proposed approach of combining in-line fluorescence and other online measurements is shown to have good potential for in-line monitoring of bioprocesses.