Multi-attribute Raman spectroscopy (MARS) for monitoring product quality attributes in formulated monoclonal antibody therapeutics

Wei, Bingchuan; Woon, Nicholas; Dai, Lu; Fish, Raphael; Tai, Michelle; Handagama, Winode; Yin, Ashley; Jia, Sun; Maier, Andrew J.; McDaniel, Dana; Kadaub, Elvira; Yang, Jessica; Saggu, Miguel; Woys, Ann Marie; Pester, Oxana; Lambert, Danny; Pell, Alex; Hao, Zhiqi; Magill, Gordon; Yim, Jack; Chan, Julia Y.; Yang, Lindsay; Macchi, Frank; Bell, Christian; Deperalta, Galahad; Chen, Yan

doi:10.1080/19420862.2021.2007564

Cited by 17 publications

(31 citation statements)

References 46 publications

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“…The concentrations of the individual species might be too low and the structural changes between differently sized species not prominent enough to be picked up by Raman spectroscopy in the short measurement times. However, Wei et al [ 37 ] showed promising results to quantify aggregates and fragments with a multi-product PLS model based on offline Raman measurements with a measurement time of 22.5 minutes.…”

Section: Resultsmentioning

confidence: 99%

Monitoring of ultra- and diafiltration processes by Kalman-filtered Raman measurements

2023

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Monitoring the protein concentration and buffer composition during the Ultrafiltration/Diafiltration (UF/DF) step enables the further automation of biopharmaceutical production and supports Real-time Release Testing (RTRT). Previously, in-line Ultraviolet (UV) and Infrared (IR) measurements have been used to successfully monitor the protein concentration over a large range. The progress of the diafiltration step has been monitored with density measurements and Infrared Spectroscopy (IR). Raman spectroscopy is capable of measuring both the protein and excipient concentration while being more robust and suitable for production measurements in comparison to Infrared Spectroscopy (IR). Regardless of the spectroscopic sensor used, the low concentration of excipients poses a challenge for the sensors. By combining sensor measurements with a semi-mechanistic model through an Extended Kalman Filter (EKF), the sensitivity to determine the progress of the diafiltration can be improved. In this study, Raman measurements are combined with an EKF for three case studies. The advantages of Kalman-filtered Raman measurements for excipient monitoring are shown in comparison to density measurements. Furthermore, Raman measurements showed a higher measurement speed in comparison to Variable Pathlength (VP) UV measurement at the trade-off of a slightly worse prediction accuracy for the protein concentration. However, the Raman-based protein concentration measurements relied mostly on an increase in the background signal during the process and not on proteinaceous features, which could pose a challenge due to the potential influence of batch variability on the background signal. Overall, the combination of Raman spectroscopy and EKF is a promising tool for monitoring the UF/DF step and enables process automation by using adaptive process control.

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Section: Resultsmentioning

confidence: 99%

Monitoring of ultra- and diafiltration processes by Kalman-filtered Raman measurements

2023

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“…With regard to PAT, spectroscopic sensors have shown to enable real‐time measurement of certain critical quality attributes (Rathore et al, 2010). Consequently, these sensors have been deployed as PAT for various unit operations in downstream processing of mAbs (Großhans et al, 2018; Rolinger et al, 2020; Wei et al, 2022). In the field of ADCs, monitoring of the conjugation reaction using UV/Vis spectra (Andris et al, 2018) and quantification of aggregation using Raman spectroscopy (Zhang et al, 2019) are recently established PAT tools.…”

Section: Introductionmentioning

confidence: 99%

An adaptive soft‐sensor for advanced real‐time monitoring of an antibody‐drug conjugation reaction

Schiemer

Weggen

Schmitt

et al. 2023

Biotech & Bioengineering

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In the production of antibody‐drug conjugates (ADCs), the conjugation reaction is a central step defining the final product composition and, hence, directly affecting product safety and efficacy. To enable real‐time monitoring, spectroscopic sensors in combination with multivariate regression models have gained popularity in recent years. The extended Kalman filter (EKF) can be used as so‐called soft‐sensor to fuse sensor predictions with long‐horizon forecasts by process models. This enables the dynamic update of the current state and provides increased robustness against experimental noise or model errors. Due to the uncertainty associated with sensor and process models in biopharmaceutical applications, the deployment of such soft‐sensors is challenging. In this study, we demonstrate the combination of an uncertainty‐aware sensor model with a kinetic reaction model using an EKF to monitor a site‐directed ADC conjugation reaction. As the sensor model, a Gaussian process regression model is presented to realize a time‐variant determination of the sensor uncertainty. The EKF fuses the time‐discrete predictions of the amount of conjugated drug from the sensor model with the time‐continuous predictions from the kinetic model. While the ADC species are not distinguishable by on‐line recorded UV/Vis spectra, the developed soft‐sensor is able to dynamically update all relevant reaction species. It could be shown that the use of time‐variant process and sensor noise computation approaches improved the performance of the EKF and achieved a reduction of the prediction error of up to 23% compared with the kinetic model. The developed framework proved to enhance robustness against noisy sensor measurements or wrong model initialization and was successfully transferred from batch to fed‐batch mode. In future, this framework could be implemented for model‐based process control and be adopted for other ADC conjugation reaction types.

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“…8 Additionally, associating specific peaks with the critical quality attributes (CQA) is intricate due to limited peak specificity and convoluted fingerprints. 10 light, both of which contribute to strong and sharp emissions in Raman spectra, with variances surpassing those from the antibody. Liquid chromatography technologies have been adapted to PAT applications by integrating with an automated sampling system for seamless online monitoring.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Raman spectroscopy experiences minimal water interference, yet protein signals are relatively weak due to their small scattering cross sections, making the analysis of dilute protein solutions challenging . Additionally, associating specific peaks with the critical quality attributes (CQA) is intricate due to limited peak specificity and convoluted fingerprints . It is crucial during spectral acquisition to shield the bioreactor from daylight and artificial light, both of which contribute to strong and sharp emissions in Raman spectra, with variances surpassing those from the antibody.…”

Section: Introductionmentioning

confidence: 99%

Continuous Online Titer Monitoring in CHO Cell Culture Supernatant Using a Herringbone Nanofluidic Filter Array

Rohskopf,

Kwon,

et al. 2023

Anal. Chem.

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Online monitoring of monoclonal antibody product titers throughout biologics process development and production enables rapid bioprocess decision-making and process optimization. Conventional analytical methods, including high-performance liquid chromatography and turbidimetry, typically require interfacing with an automated sampling system capable of online sampling and fractionation, which suffers from increased cost, a higher risk of failure, and a higher mechanical complexity of the system. In this study, a novel nanofluidic system for continuous direct (no sample preparation) IgG titer measurements was investigated. Tumor necrosis factor α (TNF-α), conjugated with fluorophores, was utilized as a selective binder for adalimumab in the unprocessed cell culture supernatant. The nanofluidic device can separate the bound complex from unbound TNF-α and selectively concentrate the bound complex for high-sensitivity detection. Based on the fluorescence intensity from the concentrated bound complex, a fluorescence intensity versus titer curve can be generated, which was used to determine the titer of samples from filtered, unpurified Chinese hamster ovary cell cultures continuously. The system performed direct monitoring of IgG titers with nanomolar resolution and showed a good correlation with the biolayer interferometry assays. Furthermore, by variation of the concentration of the indicator (TNF-α), the dynamic range of the system can be tuned and further expanded.

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Multi-attribute Raman spectroscopy (MARS) for monitoring product quality attributes in formulated monoclonal antibody therapeutics

Cited by 17 publications

References 46 publications

Monitoring of ultra- and diafiltration processes by Kalman-filtered Raman measurements

Monitoring of ultra- and diafiltration processes by Kalman-filtered Raman measurements

An adaptive soft‐sensor for advanced real‐time monitoring of an antibody‐drug conjugation reaction

Continuous Online Titer Monitoring in CHO Cell Culture Supernatant Using a Herringbone Nanofluidic Filter Array

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