Development of cell-based assay for predictively evaluating the FcγR-mediated human immune cell activation by therapeutic monoclonal antibodies

Tada, Minoru; Ishii‐Watabe, Akiko

doi:10.1016/j.bbrc.2017.02.050

Cited by 5 publications

(7 citation statements)

References 29 publications

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“…We previously reported that reporter cell lines expressing FcgRs were useful for evaluating the Fc-mediated functions of therapeutic mAbs, and the activity measured using these reporter cell lines reflected that measured using hPBMCs. 22,23 As shown in this study, such reporter cell lines are also useful for evaluating differences in FcgR-activation properties of mAb aggregates with different characteristics. Considering that FcgR activation is only part of the mechanism related to the induction of immunogenicity by mAb aggregates, further studies using hPBMCs and animals will be required to reveal the detailed relationship between FcgR-activation properties and the immunogenicity of mAb aggregates.…”

Section: Discussionmentioning

confidence: 83%

“…We previously developed FcgR-expressing reporter cell lines in which activation of FcgRs led to the expression of luciferase reporter and reported that these reporter cells could be useful for the evaluation of Fc-mediated immune cell activation by mAbs. [21][22][23] In this study, we used the FcgR-expressing reporter cell lines to examine the FcgR-activation properties of mAb aggregates with different characteristics, and we revealed that FcgR activation by mAb aggregates depends on the Fc function of the native (nonaggregated) mAbs. We also showed that mAb aggregates smaller than 1 mm can directly activate FcgRs.…”

Section: Introductionmentioning

confidence: 99%

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Fcγ Receptor Activation by Human Monoclonal Antibody Aggregates

Tada

Aoyama

Ishii‐Watabe

2020

Journal of Pharmaceutical Sciences

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Protein aggregates are a potential risk factor for immunogenicity. The measurement, characterization, and control of protein aggregates in drug products are indispensable for the development of biopharmaceuticals, including therapeutic mAbs. In this study, Fcg receptor (FcgR)-expressing reporter cell lines were used to analyze the FcgR-activation properties of mAb aggregates. Comparison of aggregates of mAbs harboring different IgG subclasses revealed that the FcgR-activation profiles of the mAb aggregates were dependent on IgG subclass. In addition, aggregates of Fc-engineered mAb with enhanced FcgRactivation properties exhibited stronger activation of FcgRs than was observed in the wild-type aggregates, whereas aggregates of Fc-engineered mAb with decreased FcgR-activation properties showed reduced activation. These results suggest that FcgR activation by mAb aggregates depends greatly on the Fc functions of the native (nonaggregated) mAbs. We also showed that aggregates of mAbs smaller than 1 mm in size have the potential to directly activate FcgRs. Unintended immune cell activation can be induced on account of FcgR activation by mAb aggregates and such FcgR activation may contribute to immunogenicity, and therefore, analysis of the FcgR-activation properties of mAb aggregates using FcgR-expressing reporter cell lines is a promising approach for the characterization of mAb aggregates.

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Section: Discussionmentioning

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Fcγ Receptor Activation by Human Monoclonal Antibody Aggregates

Tada

Aoyama

Ishii‐Watabe

2020

Journal of Pharmaceutical Sciences

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“…Studies using Fc-engineered mAbs with higher FcgR activation properties have demonstrated a positive correlation between the release of inflammatory cytokines from hPBMCs and FcgR activation. 21 We suspected that the increased secretion of cytokines from hPBMCs in the presence of SO droplets was also FcgR mediated. Nevertheless, our FcgR reporter cell assay showed similar low levels of FcgRIIA and FcgRIIIA activation in the samples regardless of the presence of SO droplets ( Supplementary Information Fig.…”

Section: Fcgr Binding By Proteins Adsorbed To So Dropletsmentioning

confidence: 99%

“…Adalimumab and etanercept were diluted to a final concentration of 10 mg/mL using 1Â PBS (pH 7.4) prepared using WFI from SO and SOF PFSs subjected to friability testing and then were added to Jurkat/Fc gamma receptors (FcgRs)/nuclear factor of activated Tcellseluciferase cells expressing FcgRIIa or FcgRIIIa. 21 We measured the resulting luciferase activities using the ONE-Glo Luciferase Assay System (Promega Corporation, Madison, WI) as described previously. 21 Control solutions prepared using nonstressed WFI were assayed identically.…”

Section: Fcgr Reporter Assaymentioning

confidence: 99%

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An Assessment of the Ability of Submicron- and Micron-Size Silicone Oil Droplets in Dropped Prefillable Syringes to Invoke Early- and Late-Stage Immune Responses

Krayukhina¹,

Yokoyama²,

Hayashihara³

et al. 2019

Journal of Pharmaceutical Sciences

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a b s t r a c tA number of biopharmaceuticals are available as lyophilized formulations along with a prefilled syringe (PFS) containing water for injection (WFI). Submicron-and micron-size droplets of lubricating silicone oil (SO) applied to the inner surface of the PFS barrel might migrate into the WFI, to which protein pharmaceuticals can adsorb, potentially inducing an immune response. In the present study, we subjected siliconized cyclo-olefin polymer PFSs filled with WFI to dropping stress to simulate actual shipping conditions as well as evaluated the risk associated with the released SO droplets. The results confirmed the undesirable effects of SO on therapeutic proteins, including adsorption to SO droplets and increased secretion of several innate cytokines from human peripheral blood mononuclear cells of a small donor panel. Assessment of immunogenicity in vivo using BALB/c mice revealed a slight increase in the plasma concentrations of antidrug antibodies over 21 days in response to SO-containing antibody samples compared to the absence of SO. These results indicate that SO droplets form complexes with pharmaceutical proteins that can potentially invoke early-and late-stage immune responses. Therefore, the use of SO-free cyclo-olefin polymer PFSs as primary containers for WFI could contribute to the enhanced safety of reconstituted biopharmaceuticals.

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