A Fully Integrated Online Platform For Real Time Monitoring Of Multiple Product Quality Attributes In Biopharmaceutical Processes For Monoclonal Antibody Therapeutics

Liu, Yang; Zhang, Chi; Chen, Jiangchao; Fernandez, Janice; Vellala, Pragna; Kulkarni, Tanmay; Aguilar, Isaiah; Ritz, Diana; Lan, Kevin; Patel, Pramthesh; Liu, Aston

doi:10.1016/j.xphs.2021.09.011

Cited by 39 publications

(39 citation statements)

References 31 publications

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“…However, we decided against the use of such modifiers to decrease the total salt concentration within our workflow that could interfere with downstream sample processing. Similar mAb binding experiments have been reported on a similar ProSIAmpler system by Liu et al 22 that also gave very similar results to when using a commercially available and pre-packed protein A columns.…”

Section: Resultssupporting

confidence: 85%

An Integrated Process Analytical Platform for Automated Monitoring of Monoclonal Antibody N-linked Glycosylation

Gyorgypal

Chundawat

2021

Preprint

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The biopharmaceutical industry is transitioning towards adoption of continuous biomanufacturing practices that are often more flexible and efficient than traditional batch processes. Regulatory agencies such as the Food and Drug Administration (FDA) are further urging use of advanced PAT to analyze the design space to increase process knowledge and enable high quality biologics production. Post-translational modification of proteins, such as N-linked glycosylation are often critical quality attributes known to affect biologics safety and efficacy hence requiring close monitoring during manufacturing. Here, we developed an online sequential-injection based PAT system, called N-GLYcanyzer, that can rapidly monitor mAb glycosylation during upstream biomanufacturing. The key innovation includes design of an integrated mAb sampling and derivation system for antibody titer and glycoform analysis in under 2 hours. The N-GLYcanyzer process includes mAb capture, deglycosylation, fluorescent glycan labeling, and glycan enrichment for direct injection and analysis on an integrated high performance liquid chromatography (HPLC) system. Different fluorescent tags and reductants were tested to maximize glycan labeling efficiency under aqueous conditions, while porous graphitized carbon (PGC) was studied for optimum glycan recovery and enrichment. We find that 2-AB labeling of glycans with 2-picoline borane as a reducing agent, using the N-GLYcanyzer workflow, gives higher glycan labeling efficiency under aqueous conditions leading to upwards of a 5-fold increase in fluorescent products intensity. Finally, we showcase how the N-GLYcanyzer platform can be implemented at/on-line to an upstream bioreactor for automated and near real-time glycosylation monitoring of a Trastuzumab biosimilar produced by Chinese Hamster Ovary (CHO) cells.Graphical AbstractN-GLYcanyzer is an automated PAT toolkit for rapid sample processing for mAb N-linked glycans analysis to enable advanced biologics manufacturing

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Section: Resultssupporting

confidence: 85%

An Integrated Process Analytical Platform for Automated Monitoring of Monoclonal Antibody N-linked Glycosylation

Gyorgypal

Chundawat

2021

Preprint

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“…2-3, 6-8 As such, the multi-attribute method (MAM), including PQA monitoring and purity testing through new peak detection (NPD), serve as an orthogonal method and could eventually replace several traditional HPLC and CE-based assays currently used for characterization and release of biopharmaceuticals. 9-19…”

Section: Introductionmentioning

confidence: 99%

“…21 The applications of using Lys-C digestion for high-resolution MS based MAM for PQA monitoring and NPD has not been reported. 9-19…”

Section: Introductionmentioning

confidence: 99%

“…[2][3][6][7][8] As such, the multi-attribute method (MAM), including PQA monitoring and purity testing through new peak detection (NPD), serve as an orthogonal method and could eventually replace several traditional HPLC and CE-based assays currently used for characterization and release of biopharmaceuticals. [9][10][11][12][13][14][15][16][17][18][19] One of the challenges for the implementation of MAM in Good Manufacturing Practice (GMP)/ quality control (QC) areas is the streamlined sample preparation, which enables robust highthroughput testing while controlling assay variability. Reduced peptide mapping-based MAM (RPM-MAM) involves enzymatic digestion of proteins into peptides for MS detection.…”

Section: Introductionmentioning

confidence: 99%

“…21 The applications of using Lys-C digestion for high-resolution MS based MAM for PQA monitoring and NPD has not been reported. [9][10][11][12][13][14][15][16][17][18][19] Beyond sample preparation and protein digestion, peptide separation is another critical aspect of MAM development with the goal of capturing as many PQAs as possible. In the past, the changes observed on heat-stressed mAbs by ion exchange chromatography (IEX) were difficult to trace back to amino acid modifications by RPM.…”

Section: Introductionmentioning

confidence: 99%

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Improvements on Sample Preparation and Peptide Separation for Reduced Peptide Mapping Based Multi-Attribute Method analysis of Therapeutic Monoclonal Antibodies Using Lys-C Digestion

Rawal

Rivera

et al. 2022

Preprint

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The mass spectrometry based multi-attribute method (MAM) has gained popularity in the field of biopharmaceutical analysis as it promises a single method for comprehensive monitoring of multiple product quality attributes (PQAs) and product purity. Sample preparation for protein digestion and peptide separation are critical considerations for a reduced peptide mapping-based MAM. To avoid desalting steps required in tryptic protein digestion and in order to improve peptide separation for hydrophilic peptides, we developed an improved robust sample preparation using Lys-C protease for high-throughput MAM testing. Additionally, this method optimizes the peptide retention and separation of a stability-indicating VSNK peptide using a HSS T3 column for comprehensive PQA monitoring. A fully automated sample preparation had similar assay variations for PQAs monitoring compared to manual sample preparation. To the best of our knowledge, this is the first report of a high-resolution MS-based MAM using Lys-C digestion with enhanced PQA monitoring for hydrophilic peptides. The improved, robust MAM workflow for protein digestion and peptide separation will pave the way for broader MAM qualification and its applications for the characterization and quality control of therapeutic monoclonal antibodies.

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Control strategy for biopharmaceutical production by model predictive control

Eslami,

Jungbauer

2024

Biotechnology Progress

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The biopharmaceutical industry is rapidly advancing, driven by the need for cutting‐edge technologies to meet the growing demand for life‐saving treatments. In this context, Model Predictive Control (MPC) has emerged as a promising solution to address the complexity of modern biopharmaceutical production processes. Its ability to optimize operations and ensure consistent product yields has made it an attractive option for manufacturers in this sector. Furthermore, MPC's alignment with the Process Analytical Technology (PAT) initiative provides an additional layer of assurance, facilitating real‐time monitoring and enabling swift adjustments to maintain process integrity. This comprehensive review delves into the various applications of MPC, ranging from robust control to stochastic model predictive control, thereby equipping biotechnologists and process engineers with a powerful toolset. By harnessing the capabilities of MPC, as elucidated in this review, manufacturers can confidently navigate the intricate bioprocessing landscape and unlock this approach's full potential in their production processes.

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A Fully Integrated Online Platform For Real Time Monitoring Of Multiple Product Quality Attributes In Biopharmaceutical Processes For Monoclonal Antibody Therapeutics

Cited by 39 publications

References 31 publications

An Integrated Process Analytical Platform for Automated Monitoring of Monoclonal Antibody N-linked Glycosylation

An Integrated Process Analytical Platform for Automated Monitoring of Monoclonal Antibody N-linked Glycosylation

Improvements on Sample Preparation and Peptide Separation for Reduced Peptide Mapping Based Multi-Attribute Method analysis of Therapeutic Monoclonal Antibodies Using Lys-C Digestion

Control strategy for biopharmaceutical production by model predictive control

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