2016 Fiscal Year Final Research Report
Development of fluorescence detection analytical ultracentrifugation
| Project/Area Number |
15K14457
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2015-04-01 – 2017-03-31
|
| Keywords | 抗体 / 複合体 / 蛋白質医薬 / 免疫原性 / 超遠心分析 / 質量分析 |
|---|
| Outline of Final Research Achievements |
We characterized the binding stoichiometry and sizes of soluble TNF-antagonist complexes for adalimumab, infliximab, and etanercept that were formed in human serum. Fluorescence-detected sedimentation velocity analytical ultracentrifugation analyses revealed that adalimumab and infliximab formed a range of complexes with TNF, with the major complexes consisting of 3 molcules of the respective antagonist and one or 2 molcules of TNF. Considerably greater amounts of high-molecular-weight complexes were detected for infliximab in human serum. Etanerept exclusively formed 1:1 complexes with TNF in PBS, and a small amount of complexes with higher stoichiometry was detected in human serum. Consistent with these biophysical characterizations, a reporter assay showed that adalimumab and infliximab, but not etanercept, exerted FcgRIIa- and FcgRIIIa-mediated cell signaling in the presence of TNF and that infliximab exhibited higher potency than adalimumab.
|
| Free Research Field |
生物物理化学
|
