Monitoring of Flow‐Induced Aggregation and Conformational Change of Monoclonal Antibodies

Wolfrum, Sonja; Weichsel, Ulrike; Siedler, Michael; Weber, Carsten

doi:10.1002/cite.201600161

Cited by 4 publications

(3 citation statements)

References 18 publications

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“…There is thus a need to develop stress tests that more closely replicate the conformational ensemble generated during processing and transport. In light of this, we and others have shown that hydrodynamic extensional flow fields encountered during the nano-filtration, pumping, and fill-finish steps of bio-processing can trigger protein aggregation (Charm & Wong, 1981;Dobson et al, 2017;Simon, Krause, Weber, & Peukert, 2011;Wolfrum, Weichsel, Siedler, Weber, & Peukert, 2017). Using a reciprocating extensional and shear flow device (EFD) (Figure 1a), we showed that extensional flow fields can induce the conformational unfolding/remodeling of bovine serum albumin (BSA, Figure 1b), leading to aggregation that was characterized and quantified by an array of biophysical techniques including DLS, NTA, and TEM (Dobson et al, 2017).…”

Section: Introductionmentioning

confidence: 93%

Using extensional flow to reveal diverse aggregation landscapes for three IgG1 molecules

Willis

Kumar

Dobson

et al. 2018

Biotech & Bioengineering

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Monoclonal antibodies (mAbs) currently dominate the biopharmaceutical sector due to their potency and efficacy against a range of disease targets. These proteinaceous therapeutics are, however, susceptible to unfolding, mis‐folding, and aggregation by environmental perturbations. Aggregation thus poses an enormous challenge to biopharmaceutical development, production, formulation, and storage. Hydrodynamic forces have also been linked to aggregation, but the ability of different flow fields (e.g., shear and extensional flow) to trigger aggregation has remained unclear. To address this question, we previously developed a device that allows the degree of extensional flow to be controlled. Using this device we demonstrated that mAbs are particularly sensitive to the force exerted as a result of this flow‐field. Here, to investigate the utility of this device to bio‐process/biopharmaceutical development, we quantify the effects of the flow field and protein concentration on the aggregation of three mAbs. We show that the response surface of mAbs is distinct from that of bovine serum albumin (BSA) and also that mAbs of similar sequence display diverse sensitivity to hydrodynamic flow. Finally, we show that flow‐induced aggregation of each mAb is ameliorated by different buffers, opening up the possibility of using the device as a formulation tool. Perturbation of the native state by extensional flow may thus allow identification of aggregation‐resistant mAb candidates, their bio‐process parameters and formulation to be optimized earlier in the drug‐discovery pipeline using sub‐milligram quantities of material.

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Section: Introductionmentioning

confidence: 93%

Using extensional flow to reveal diverse aggregation landscapes for three IgG1 molecules

Willis

Kumar

Dobson

et al. 2018

Biotech & Bioengineering

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“…The potential for hydrodynamic forces to induce mAb aggregation has long been of concern, with fresh attention being paid recently . Our efforts to address this inspired design of the EFD and its application to three IgG1s .…”

Section: Discussionmentioning

confidence: 99%

“…The potential for hydrodynamic forces to induce mAb aggregation has long been of concern, 9 with fresh attention being paid recently. 77 , 78 , 79 , 80 Our efforts to address this inspired design of the EFD and its application to three IgG1s. 59 , 60 With the above in mind, we set out to analyze the extent of EFD‐induced aggregation for a subset of 33 mAbs from the Jain et al dataset.…”

Section: Discussionmentioning

confidence: 99%

The uniqueness of flow in probing the aggregation behavior of clinically relevant antibodies

Willis

Kumar

Jain

et al. 2020

Engineering Reports

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The development of therapeutic monoclonal antibodies (mAbs) can be hindered by their tendency to aggregate throughout their lifetime, which can illicit immunogenic responses and render mAb manufacturing unfeasible. Consequently, there is a need to identify mAbs with desirable thermodynamic stability, solubility, and lack of self-association. These behaviors are assessed using an array of in silico and in vitro assays, as no single assay can predict aggregation and developability. We have developed an extensional and shear flow device (EFD), which subjects proteins to defined hydrodynamic forces which mimic those experienced in bioprocessing. Here, we utilize the EFD to explore the aggregation propensity of 33 IgG1 mAbs, whose variable domains are derived from clinical antibodies. Using submilligram quantities of material per replicate, wide-ranging EFD-induced aggregation (9-81% protein in pellet) was observed for these mAbs, highlighting the EFD as a sensitive method to assess aggregation propensity. By comparing the EFD-induced aggregation data to those obtained previously from 12 other biophysical assays, we show that the EFD provides distinct information compared with current measures of adverse biophysical behavior. Assessing a candidate's liability to hydrodynamic force thus adds novel insight into the rational selection of developable mAbs that complements other assays. K E Y W O R D S aggregation, developability, extensional flow, monoclonal antibody, shear flowThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Characterization of Grinding-Induced Subvisible Particles and Free Radicals in a Freeze-Dried Monoclonal Antibody Formulation

et al. 2022

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Monitoring of Flow‐Induced Aggregation and Conformational Change of Monoclonal Antibodies

Cited by 4 publications

References 18 publications

Using extensional flow to reveal diverse aggregation landscapes for three IgG1 molecules

Using extensional flow to reveal diverse aggregation landscapes for three IgG1 molecules

The uniqueness of flow in probing the aggregation behavior of clinically relevant antibodies

Characterization of Grinding-Induced Subvisible Particles and Free Radicals in a Freeze-Dried Monoclonal Antibody Formulation

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