RRC ID 45035
著者 Tada M, Ishii-Watabe A, Suzuki T, Kawasaki N.
タイトル Development of a cell-based assay measuring the activation of FcγRIIa for the characterization of therapeutic monoclonal antibodies.
ジャーナル PLoS One
Abstract Antibody-dependent cellular cytotoxicity (ADCC) is one of the important mechanisms of action of the targeting of tumor cells by therapeutic monoclonal antibodies (mAbs). Among the human Fcγ receptors (FcγRs), FcγRIIIa is well known as the only receptor expressed in natural killer (NK) cells, and it plays a pivotal role in ADCC by IgG1-subclass mAbs. In addition, the contributions of FcγRIIa to mAb-mediated cytotoxicity have been reported. FcγRIIa is expressed in myeloid effector cells including neutrophils and macrophages, and it is involved in the activation of these effector cells. However, the measurement of the cytotoxicity via FcγRIIa-expressing effector cells is complicated and inconvenient for the characterization of therapeutic mAbs. Here we report the development of a cell-based assay using a human FcγRIIa-expressing reporter cell line. The FcγRIIa reporter cell assay was able to estimate the activation of FcγRIIa by antigen-bound mAbs by a very simple method in vitro. The usefulness of this assay for evaluating the activity of mAbs with different abilities to activate FcγRIIa was confirmed by the examples including the comparison of the activity of the anti-CD20 mAb rituximab and its Fc-engineered variants, and two anti-EGFR mAbs with different IgG subclasses, cetuximab (IgG1) and panitumumab (IgG2). We also applied this assay to the characterization of a force-oxidized mAb, and we observed that oxidation significantly decreased the FcγRIIa activation by EGFR-bound cetuximab. These results suggest that our FcγRIIa reporter assay is a promising tool for the characterization of therapeutic mAbs, including Fc-engineered mAbs, IgG2-subclass mAbs, and their product-related variants.
巻・号 9(4)
ページ e95787
公開日 2014-1-1
DOI 10.1371/journal.pone.0095787
PII PONE-D-14-00922
PMID 24752341
PMC PMC3994145
MeSH Antibodies, Monoclonal / therapeutic use* Antibodies, Monoclonal, Humanized / therapeutic use Antibodies, Monoclonal, Murine-Derived / therapeutic use Biological Assay / methods* Cell Line Cetuximab Flow Cytometry Humans Jurkat Cells Panitumumab Receptors, IgG / metabolism* Rituximab
IF 2.74
引用数 33
WOS 分野 IMMUNOLOGY
リソース情報
ヒト・動物細胞 Jurkat(RCB0806)