In‐line monitoring of protein concentration with MIR spectroscopy during UFDF

Abstract

Rapid increase of product titers in upstream processes has presented challenges for downstream processing, where purification costs increase linearly with the increase of the product yield. Hence, innovative solutions are becoming increasingly popular. Process Analytical Technology (PAT) tools, such as spectroscopic techniques, are on the rise due to their capacity to provide real-time, precise analytics. This ensures consistent product quality and increased process understanding, as well as process control. Mid-infrared spectroscopy (MIR) has emerged as a highly promising technique within recent years, owing to its ability to monitor several critical process parameters at the same time and unchallenging spectral analysis and data interpretation. For in-line monitoring, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR) is a method of choice, as it enables reliable measurements in a liquid environment, even though water absorption bands are present in the region of interest. Here, we present MIR spectroscopy as a monitoring tool of critical process parameters in ultrafiltration/diafiltration (UFDF). MIR spectrometer was integrated in the UFDF process in an in-line fashion through a single-use flow cell containing a single bounce silicon ATR crystal. The results indicate that the one-point calibration algorithm applied to the MIR spectra, predicts highly accurate protein concentrations, as compared with validated offline analytical methods.

An adaptive modeling approach using spiking‐augmentation method to improve chemometric model performance in bioprocess monitoring

“…The FTIR spectra of mAbs consist of well-defined peaks that represent protein structural information, which has been reported for quantitative analysis in literature. 12,[28][29][30] Successful at-line and in-line applications has been reported using IR techniques to quantify product titer, aggregation, and host cell proteins in downstream unit operations. [31][32][33] Multivariate data analysis is often required to understand and correlate the FTIR spectra to a quality attribute or process parameter, such as product titer, via the chemometric modeling of a calibration data set.…”

“…Fourier transform infrared (FTIR) spectroscopy, utilizing the electromagnetic radiation region of 800–4000 cm −1 wavenumber is a promising candidate among all the technologies. The FTIR spectra of mAbs consist of well‐defined peaks that represent protein structural information, which has been reported for quantitative analysis in literature 12,28–30 . Successful at‐line and in‐line applications has been reported using IR techniques to quantify product titer, aggregation, and host cell proteins in downstream unit operations 31–33 .…”