In situ cell differentiation monitoring of Catharanthus roseus suspension culture processes by NIR spectroscopy

Bioprocess and Biosystems Engineering - Plant suspension culture is attracting interest as a promising platform to produce biological medicines due to the absence of virus, prions or DNA related to...     

D-Aminoacylase from a novel producer: Stenotrophomonas maltophilia ITV-0595

A novel bacterial strain producing D-aminoacylase was isolated from organic waste and identified as Stenotrophomonas maltophilia ITV-0595. The isolation was performed using N-acetyl-D-phenylglycine (NAcDPG) as the sole source of C and N. The optimum pH for enzyme expression was 8 at 37°C. Using N-Ac-DPG concentrations from 0.5 up to 3% w/v, it was observed that at the 1% level, the microorganism showed acceptable responses in both enzyme activities and cell growth. From the different tested compounds N-acetyl-D-methionine (1%) was the best enzyme inducer (Sp. act. = 4.14 U mg⁻¹ protein, Vol. act. = 0.17 U ml⁻¹) and the only one that increased cell growth. Received 13 June 1997/ Accepted in revised form 29 October 1998     

Interest of locally weighted regression to overcome nonlinear effects during in situ NIR monitoring of CHO cell culture parameters and antibody glycosylation

Animal cell culture processes have become the standard platform to produce therapeutic proteins such as recombinant monoclonal antibodies (mAb). Since the mAb quality could be subject to significant changes depending on manufacturing process conditions, real time monitoring and control systems are required to ensure mAb specifications mainly glycosylation and patient safety. Up to now, real time monitoring glycosylation of proteins has received scarce attention. In this article, the use of near infrared (NIR) to monitor mAb glycosylation has been reported for the first time. Whereas monitoring models are mainly constructed using linear partial least squares regressions (PLSR), evidences presented in this study indicate nonlinearity relationship between in situ captured spectra and compound concentrations, compromising the PLSR performances. A novel and simple approach was proposed to fit nonlinearity using the locally weighted regression (LWR). The LWR models were found to be more appropriate for handling information contained in spectra so that real time monitoring of cultures were accurately performed. Moreover, for the first time, the LWR calibration models allowed mAb glycosylation to be monitored, in a real time manner, by using in situ NIR spectroscopy. These results represent a further step toward developing active-control feedback of animal cell processes, particularly for ensuring properties of biologics.