ID 112359
著者
Kimura, Hitoshi Kyoto Pharmaceutical University|Tokushima University
Mikawa, Shiho Kyoto Pharmaceutical University|Tokushima University
Mizuguchi, Chiharu Kyoto Pharmaceutical University|Tokushima University
Horie, Yuki Kobe Pharmaceutical University
Morita, Izumi Kobe Pharmaceutical University
Oyama, Hiroyuki Kobe Pharmaceutical University
Ohgita, Takashi Kyoto Pharmaceutical University
Takeuchi, Atsuko Kobe Pharmaceutical University
Lund-Katz, Sissel University of Pennsylvania
Akaji, Kenichi Kyoto Pharmaceutical University
Kobayashi, Norihiro Kobe Pharmaceutical University
斎藤, 博幸 Kyoto Pharmaceutical University KAKEN研究者をさがす
資料タイプ
学術雑誌論文
抄録
Apolipoprotein A-I (apoA-I) undergoes a large conformational reorganization during remodeling of high-density lipoprotein (HDL) particles. To detect structural transition of apoA-I upon HDL formation, we developed novel monoclonal antibodies (mAbs). Splenocytes from BALB/c mice immunized with a recombinant human apoA-I, with or without conjugation with keyhole limpet hemocyanin, were fused with P3/NS1/1-Ag4-1 myeloma cells. After the HAT-selection and cloning, we established nine hybridoma clones secreting anti-apoA-I mAbs in which four mAbs recognize epitopes on the N-terminal half of apoA-I while the other five mAbs recognize the central region. ELISA and bio-layer interferometry measurements demonstrated that mAbs whose epitopes are within residues 1–43 or 44–65 obviously discriminate discoidal and spherical reconstituted HDL particles despite their great reactivities to lipid-free apoA-I and plasma HDL, suggesting the possibility of these mAbs to detect structural transition of apoA-I on HDL. Importantly, a helix-disrupting mutation of W50R into residues 44–65 restored the immunoreactivity of mAbs whose epitope being within residues 44–65 against reconstituted HDL particles, indicating that these mAbs specifically recognize the epitope region in a random coil state. These results encourage us to develop mAbs targeting epitopes in the N-terminal residues of apoA-I as useful probes for monitoring formation and remodeling of HDL particles.
掲載誌名
Scientific Reports
ISSN
20452322
出版者
Springer Nature
7
開始ページ
2988
発行日
2017-06-07
備考
Supplementary information : srep_7_2988_s1.pdf
権利情報
© The Author(s) 2017
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出版社版DOI
出版社版URL
フルテキストファイル
言語
eng
著者版フラグ
出版社版
部局
医学系
薬学系