Multi‐cell‐line learning for the data‐driven construction of mechanistic metabolic models DOI Creative Commons
Yen‐An Lu,

Meghan G. McCann,

Wei‐Shou Hu

et al.

Biotechnology and Bioengineering, Journal Year: 2024, Volume and Issue: 121(9), P. 2833 - 2847

Published: June 3, 2024

Mammalian cells are commonly used as hosts in cell culture for biologics production the pharmaceutical industry. Structured mechanistic models of metabolism have been to capture complex cellular mechanisms that contribute varying metabolic shifts different lines. However, little research has focused on impact temporal changes enzyme abundance and activity modeling metabolism. In this work, we present a framework constructing integrate growth-signaling control transcript dynamics. The proposed approach is applied build three Chinese hamster ovary (CHO) lines using fed-batch data time-series profiles. Leveraging information from transcriptome data, develop parameter estimation based multi-cell-line (MCL) learning, which combines sets trains individual cell-line jointly improve model accuracy. computational results demonstrate important role growth signaling variability well virtue MCL with limited amount data. resulting exhibit high level accuracy predicting distinct behaviors lines; these can potentially be accelerate process development biomanufacturing new protein therapeutics.

Language: Английский

Approved delivery strategies for biopharmaceuticals DOI
Makhloufi Zoulikha, Zhongjian Chen, Jun Wu

et al.

Chinese Chemical Letters, Journal Year: 2024, Volume and Issue: 36(2), P. 110225 - 110225

Published: July 8, 2024

Language: Английский

Citations

7
Affinity chromatography for virus-like particle manufacturing: Challenges, solutions, and perspectives DOI
Jing Ma, Zengquan Tian, Qinghong Shi

et al.

Journal of Chromatography A, Journal Year: 2024, Volume and Issue: 1721, P. 464851 - 464851

Published: March 27, 2024

Language: Английский

Citations

6
Impact of IgG subclass on monoclonal antibody developability DOI Creative Commons

Paul Cain,

Lihua Huang, Yu Tang

et al.

mAbs, Journal Year: 2023, Volume and Issue: 15(1)

Published: March 21, 2023

IgG-based monoclonal antibody therapeutics, which are mainly IgG1, IgG2, and IgG4 subclasses or related variants, have dominated the biotherapeutics field for decades. Multiple laboratories reported that IgG possess different molecular characteristics can affect their developability. For example, most popular subclass is known to a characteristic degradation pathway its hinge fragility. However, there remains paucity of studies systematically evaluate on manufacturability long-term stability. We thus conducted systematic study 12 mAbs derived from three sets unrelated variable regions, each cloned into an IgG1 variant with diminished effector functions, stabilized further reduced FcγR interaction, impact high concentration stability in common formulation buffer matrix. Our evaluation included Chinese hamster ovary cell productivity, host protein removal efficiency, N-linked glycan structure at conserved N297 Fc position, solution appearance concentration, aggregate growth, fragmentation, charge profile change, post-translational modification upon thermal stress conditions storage refrigerated temperature. results elucidated attributes all subclasses, as well those unique certain domains, providing new insight effects These learnings be used enable balanced decision selection therapeutic antibodies aid acceleration product development process.

Language: Английский

Citations

12
Exploring the sustainability of single use plastics in the biopharmaceuticals sector: a scoping review of challenges, opportunities, and impacts DOI Creative Commons

Malcolm Goggin,

Aysha Fleming, Cormac MacMahon

et al.

Frontiers in Sustainability, Journal Year: 2025, Volume and Issue: 6

Published: March 10, 2025

Single-use plastics (SUPs) are synonymous with the biopharmaceuticals sector, facilitating economies of scale, process efficiency, flexibility and sterility assurance, all a seemingly negligible environmental footprint. Yet, in ever-tightening regulation, mandated by Sustainable Development Goals (SDGs) concern for large-scale industrial impacts, sustainability SUP consumption is increasingly being questioned. Whilst sector contributes to human welfare, its transition risk unlikely remain immune societal pressure more sustainable production. This article aims present scoping review apparent contradiction between sectoral adoption increasing importance circularity. The approach relies on three interwoven strands evidence: [i] intersectionality policy regulation biopharmaceuticals, [ii] single-use technology [iii] applications circular economy principles technology. It argued that, whilst life-cycle analysis (LCA) SUPs articulates an benefit vis-à-vis conventional technology, high energy intensity embodied carbon stainless steel renders comparison redundant. Moreover, there dearth evidence circularity, post-use, end-of-life considerations. Likewise, appears be little sector-wide appetite embryonic solutions enhancing such as biodegradables, offsets, reusability, waste-to-energy, ocean cleanup. Urgent mission-driven research required LCA, business model feasibility, materials innovation, regulatory frameworks, sectoral-wide impact. A design-driven inquisition their interactions, based symbiosis, could inform potential pathways.

Language: Английский

Citations

0
Scalable process development for rAAV transient transfection production using computational fluid dynamics modeling DOI
Jianfa Ou, Yawen Tang,

Alexander Williams

et al.

Biotechnology Progress, Journal Year: 2025, Volume and Issue: unknown

Published: April 4, 2025

Recombinant adeno-associated virus (rAAV) is a promising delivery vehicle for cell and gene therapies. Upstream development faces challenges like low productivity inconsistent performance despite advancements. This study presents scale-up design robust rAAV production at 250 L scale using transfection system. Initial process in shake flasks optimized plasmid ratio to improve production. However, genome titer decreased by up 50% stirred-tank bioreactors, likely due mechanical shear forces. Stirred-tank bioreactors were modeled with computational fluid dynamics (CFD) M-STAR (250 mL, 5 L, 50 L) empirical correlations Dynochem L). Hydrodynamics characterized provide normalized stress across different geometries. The power per unit volume (P/V) of 71 W/m3 was optimal the mL bioreactor, focusing on growth, titer, capsid full ratio. Based CFD modeling, P/V 20 projected perform best scales during development, confirmed comparable flask culture. A 15 subsequently final scale. negative impact could be further mitigated adding extra Poloxamer-188 as protectant. Additionally, pre-transfection viable density (VCD) identified critical attribute. included 30% fixed-volume dilution culture along controlled DNA complexation conditions robustness. Sequential demonstrated consistent growth

Language: Английский

Citations

0
Pharmaceutical Analysis of Protein–Peptide Coformulations and the Influence of Polysorbates DOI Creative Commons
Joseph Whiteley, Susanna Abrahmsén‐Alami,

Jonathan Booth

et al.

Molecular Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown

Published: April 30, 2025

Coformulation is an approach to formulating multiple biopharmaceutical therapeutics in a single formulation, promising the benefits of both therapies one dose. However, as molecular stability key consideration traditional formulations, coformulations will require extensive investigation. This study evaluated effects formulation stabilizers, specifically surfactants, at different grades, namely, regular grade (RG) Tween 20 and 80 Super-refined Polysorbate 80. Their were assessed through their interactions with human serum albumin (HSA) glucagon-like peptide-1 (GLP-1) receptor agonist (MEDI7219). Isothermal titration calorimetry (ITC) differential scanning (DSC) implemented determine strength binding thermal tertiary system. ITC confirmed that upon MEDI7219 into solution HSA RG, affinity peptide was reduced, resulting negatively cooperative binding. when titrated grades 80, increased positive DSC established similar extent polysorbates. Combining polysorbate did not further increase HSA; however, it reduce unfolding molecules absence heat. Overall, unique findings this have demonstrated order addition ternary coformulation affects final composition which important for pharmaceutical development.

Language: Английский

Citations

0
A comparison of SWATH-MS methods for measurement of residual host cell proteins in adeno-associated virus preparations DOI Creative Commons

Thomas M. Leibiger,

Lie Min,

Kelvin H. Lee

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: May 2, 2025

Introduction Analysis of residual host cell proteins in adeno-associated virus (AAV) preparations is challenging due to low availability and high complexity samples. One strategy address these challenges through development improved liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods with greater sensitivity reduced sample requirement. Methods In this work, we compare the performance four sequential window acquisition all theoretical fragment ion spectra (SWATH-MS) for identification quantitation HCPs rAAV2, -5, -8, -9 produced human embryonic kidney 293 (HEK293) cells purified using immunoaffinity chromatography. Key SWATH-MS parameters including spectral library construction (data dependent vs. silico ), data processing software (DIA-NN Skyline), spectrometer instrument (Sciex TripleTOF 6600 Sciex ZenoTOF 7600) were assessed. Method attributes requirement time, method outputs protein precursor identifications, comparisons across methods, coefficients variance (CV) considered help establish a workflow well-suited rAAV HCP analytics. Results A 78% increase 80% reduction requirement, 70% runtime was achieved an library, DIA-NN, collection 7600 (DIA-NN-7600 method) compared previously established DDA-derived Skyline, (Skyline-DDA-6600 method). Additionally, DIA-NN-7600 shows median CV below 10% triplicate acquisitions, comparable other panel highly abundant identified downstream processing. Discussion This work highlights specifically tailored analysis.

Language: Английский

Citations

0
Behavior of host-cell-protein-rich aggregates in antibody capture and polishing chromatography DOI Creative Commons
Chase E. Herman,

Lie Min,

Leila H. Choe

et al.

Journal of Chromatography A, Journal Year: 2023, Volume and Issue: 1702, P. 464081 - 464081

Published: May 18, 2023

Language: Английский

Citations

10
Bayesian cell therapy process optimization DOI
Evan Claes, Tommy Heck,

Kathleen Coddens

et al.

Biotechnology and Bioengineering, Journal Year: 2024, Volume and Issue: 121(5), P. 1569 - 1582

Published: Feb. 19, 2024

Abstract Optimizing complex bioprocesses poses a significant challenge in several fields, particularly cell therapy manufacturing. The development of customized, closed, and automated processes is crucial for their industrial translation addressing large patient populations at sustainable price. Limited understanding the underlying biological mechanisms, coupled with highly resource‐intensive experimentation, are two contributing factors that make these next‐generation challenging. Bayesian optimization (BO) an iterative experimental design methodology addresses challenges, but has not been extensively tested situations require parallel experimentation variability. In this study, we present evaluation noisy, BO increasing noise levels batch sizes on silico bioprocesses, compare it to industry state‐of‐the‐art. As vitro showcase, apply method monocyte purification unit operation. results show significantly outperforms state‐of‐the‐art, requiring approximately 50% fewer experiments average. This study highlights potential as valuable tool process optimization.

Language: Английский

Citations

3
Analytical characterization of host‐cell‐protein‐rich aggregates in monoclonal antibody solutions DOI
Chase E. Herman,

Lie Min,

Leila H. Choe

et al.

Biotechnology Progress, Journal Year: 2023, Volume and Issue: 39(4)

Published: April 5, 2023

Host-cell proteins (HCPs) and high molecular weight (HMW) species have historically been treated as independent classes of impurities in the downstream processing monoclonal antibodies (mAbs), but recent indications suggest that they may be partially linked. We explored this connection with a shotgun proteomic analysis HMW were isolated from harvest cell culture fluid (HCCF) protein A eluate using size-exclusion chromatography (SEC). As part analysis, cross-digest study was performed which samples analyzed both standard native digest techniques to enable fair comparison between bioprocess pools. This reveals HCP profiles HCCF overlap substantially more than previous work has suggested, because hundreds HCPs are conserved aggregates up ~50 nm hydrodynamic radius persist through capture step. Quantitative SWATH proteomics suggests majority eluate's mass is found such aggregates, corroborated by ELISA measurements on SEC fractions. The data also show intra-aggregate concentrations individual positively correlated eluate, generally considered difficult remove tend concentrated their counterparts. These observations support prior hypotheses regarding aggregate-mediated persistence highlight importance mechanism.

Language: Английский

Citations

8