ミヤノ　ケイ   Kei Miyano   宮野　佳    所属   川崎医科大学  医学部 一般教養　自然科学    職種   講師
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	Fine definition of the epitopes on the human gp91phox/NOX2 for the monoclonal antibodies CL-5 and 48
掲載誌名	正式名：Journal of Immunological Methods 略　 称：J Immunol Methods ISSNコード：00221759
掲載区分	国外
巻・号・頁	501
著者・共著者	Kawai C, Miyano K, Okamoto S, Yamauchi A, Kuribayashi F.
発行年月	2022/02
概要	Superoxide-producing NADPH oxidase, gp91phox/NOX2, in phagocytes plays a critical role in the host defenses against pathogens. Moreover, gp91phox/NOX2 contributes to the oxidative stress in endothelial cells. Therefore, investigating the level of gp91phox/NOX2 with immunoblotting is important for estimating the amount of superoxide produced. Here, we showed that the epitopes in human gp91phox/NOX2 recognized by monoclonal antibodies (mAbs) CL-5 and 48 were in amino acids 132–147 and 136–144, respectively. Although the epitopes were close to the N-glycosylation sites, N-glycan maturation did not affect mAbs recognition. When Pro-136 was substituted with Arg, the corresponding mouse residue, human gp91phox/NOX2 was not recognized by mAbs CL-5 and 48; however, the substitution did not affect gp91phox/NOX2-based oxidase activity. This finding explains why these mAbs specifically recognize the human but not mouse gp91phox/NOX2. Hence, these mAbs are useful for investigating the level of gp91phox/NOX2 without amino acid substitutions in the epitopes.
DOI	https://doi.org/10.1016/j.jim.2021.113213
PMID	34971634