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Researchers
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KURIHARA Yasuyuki
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【Published Thesis】 Coxfa4l3, a novel mitochondrial electron transport chain Complex 4 subunit protein, switches from Coxfa4 during spermatogenesis. 2020.02
【Published Thesis】 Rapid, simple, and effective strategy to produce monoclonal antibodies targeting protein structures using hybridoma technology(Journal of Biological Engineering) 2023.03
【Published Thesis】 Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody.(J.Biosci. Bioeng.) 2021.02
【Published Thesis】 Rapid, simple, and effective strategy to produce monoclonal antibodies targeting protein structures using hybridoma technology(Journal of Biological Engineering) 2023.03
【Published Thesis】 Ectopic expression of the mitochondrial protein COXFA4L3 in human sperm acrosome and its potential application in the selection of male infertility treatments(Reproductive Medicine and Biology) 2024
【Published Thesis】 Rapid and reliable hybridoma screening method that is suitable for production of functional structure-recognizing monoclonal antibody. 2021.02
【Published Thesis】 Monoclonal antibody production technology optimised for cytometry: from theory to practice.(Cytometry Research) 2024.09
Kobe University Doctor Course Completed
Toho University Master Course Completed
Toho University Department of Biology Graduated
Duty Yokohama National UniversityFaculty of Engineering Division of Materials Science and Chemical Engineering Professor
Duty Yokohama National UniversityResearch Institute of Environment and Information Sciences Professor
Duty Yokohama National UniversityResearch Institute of Environment and Information Sciences Associate Professor
Duty Yokohama National UniversityResearch Institute of Environment and Information Sciences Associate Professor
Duty Yokohama National UniversityResearch Institute of Environment and Information Sciences Research Assistant
University of Pennsylvania, Center for Research on Reproduction and Women's Health, Visiting Faculty Research Associate
National Institute of Radiological Sciences, Department of Biology Researcher
Japan Society for the Promotion of Science, Research Fellow
Life Science / Molecular biology
ミトコンドリア電子伝達系の活性制御
Project Year：
迅速簡便なモノクローナル抗体作製法の開発
Project Year：
哺乳動物の生殖細胞の発生分化に関わるRNA情報の発現制御の研究
Project Year：
小笠原100の素顔Ⅱ
東京農大小笠原100の素顔編集委員会（ Role： Other）
東京農大出版会 （ ISBN:4-88694-042-0 ）
Language：Japanese Book type：General book, introductory book for general audience
制限酵素、「ヒドロラーゼ」
栗原靖之、上杉晴一（ Role： Joint author）
廣川書店
Language：Japanese Book type：Scholarly book
高感度蛍光色素を使ったポストステイニングによる蛍光ディファレンシャルディスプレイ(SSDD)法、「non‐RI実験の最新プロトコール 蛍光の原理と実際:遺伝子解析からバイオイメージングまで」
丸本佳代子、栗原靖之、（ Role： Joint author）
羊土社
Language：Japanese Book type：Scholarly book
タンパク質と核酸のゲル電気泳動の高感度蛍光染色剤を使った染色法、「non‐RI実験の最新プロトコール 蛍光の原理と実際:遺伝子解析からバイオイメージングまで」
明神玲子、栗原靖之、（ Role： Joint author）
羊土社
Language：Japanese Book type：Scholarly book
Genetics in Wild Mice, Its Application to Biomedical Research
H. Suzuki and Y. Kurihara （ Role： Joint author）
Japan Scientific Societies Press
Language：Japanese Book type：Scholarly book
Atsumi Sakaguchi, Yoichiro Tanaka, Eiki Shoji, Teppei Takeshima, Rina Sakamaki, Takao Matsuba and … Show more authors
Atsumi Sakaguchi, Yoichiro Tanaka, Eiki Shoji, Teppei Takeshima, Rina Sakamaki, Takao Matsuba and Yasuyuki Kurihara Hide authors
Journal of Biological Engineering 17 ( 1 ) 2023.3 [Reviewed]
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Springer Nature Joint Work
BACKGROUND: Monoclonal antibodies are essential in life science research and developing antibody drugs and test drugs. Various methods have been developed to obtain monoclonal antibodies, among which hybridoma technology continues to be widely used. However, developing a rapid and efficient method for obtaining conformation-specific antibodies using hybridoma technology remains challenging. We previously developed the membrane-type immunoglobulin-directed hybridoma screening (MIHS) method, which is a flow cytometry-based screening technique based on the interaction between the B-cell receptor expressed on the hybridoma cell surface and the antigen protein, to obtain conformation-specific antibodies.
RESULTS: In this study, we proposed a streptavidin-anchored ELISA screening technology (SAST) as a secondary screening method that retains the advantages of the MIHS method. Anti-enhanced green fluorescent protein monoclonal antibodies were generated as a model experiment, and their structural recognition abilities were examined. Examination of the reaction profiles showed that all monoclonal antibodies obtained in this study recognize the conformational epitopes of the protein antigen. Furthermore, these monoclonal antibodies were classified into two groups: those with binding activities against partially denatured proteins and those with complete loss of binding activities. Next, when screening monoclonal antibodies by the MIHS method as the first screening, we found that monoclonal antibodies with stronger binding constants may be selected by double-staining for hybridomas with fluorescently labeled target antigens and fluorescently labeled B cell receptor antibodies.
CONCLUSIONS: The proposed two-step screening method, which incorporates MIHS and SAST, constitutes a rapid, simple, and effective strategy to obtain conformation-specific monoclonal antibodies generated through hybridoma technology. The novel monoclonal antibody screening strategy reported herein could accelerate the development of antibody drugs and antibody tests.
Other Link： https://link.springer.com/article/10.1186/s13036-023-00345-9
Atsumi Sakaguchi, Chika Nakajima, Ayuko Sawano, Yoichiro Tanaka, and Yasuyuki Kurihara
J.Biosci. Bioeng. 131 ( 6 ) 696 - 702 2021.2 [Reviewed]
DOI Web of Science PubMed CiNii Research
Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier Joint Work
Monoclonal antibodies are extremely valuable functional biomaterials that are widely used not only in life science research but also in antibody drugs and test drugs. There is also a strong need to develop high-quality neutralizing antibodies as soon as possible in order to stop the rapid spread of new infectious diseases such as the SARS-CoV-2 virus. This study has developed a membrane-type immunoglobulin-directed hybridoma screening (MIHS) method for obtaining high-quality monoclonal antibodies with high efficiency and high speed. In addition to these advantages, this paper demonstrates that the MIHS method can selectively obtain monoclonal antibodies that specifically recognize the functional structure of proteins. The MIHS method is a useful technology that greatly contributes to the research community because it can be easily introduced in any laboratory that uses a flow cytometer.
Masahiro Endou, Kaito Yoshida, Makoto Hirota, Chika Nakajima, Atsumi Sakaguchi, Naoto Komatsubara,Y … Show more authors
Masahiro Endou, Kaito Yoshida, Makoto Hirota, Chika Nakajima, Atsumi Sakaguchi, Naoto Komatsubara,Yasuyuki Kurihara Hide authors
Mitochondrion 52 1 - 7 2020.2 [Reviewed]
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier Joint Work
We identified Coxfa4l3, previously called C15orf48 or Nmes1, as a novel accessory protein of Complex IV of the mitochondrial electron transport chain (ETC). Amino acid sequence comparison, the intracellular localization and the protein expression data showed that the protein is the third isoform of Coxfa4 and the expression of Coxfa4 and Coxfa4l3 proteins during spermatogenesis showed a mutually exclusive pattern, implying that Coxfa4 replaces Coxfa4l3 in Complex IV after meiosis. These results may provide some insight into the unique mechanism of ATP production in late spermatogenesis.
Flow cytometry-based method for rapid and high-throughput screening of hybridoma cells secreting monoclonal antibody
赤城幸、中嶋千佳、田中陽一郎、栗原靖之
Journal of Bioscience and Bioengeering 125 ( 4 ) 464 - 469 2018.5 [Reviewed]
Language：English Publishing type：Research paper (scientific journal) Joint Work
Ectopic expression of the mitochondrial protein COXFA4L3 in human sperm acrosome and its potential application in the selection of male infertility treatments
Yusuke Fujisawa, Sayaka Kikuchi, Fujino Kuba, Kosei Oishi, Soushi Murayama, Tomoya Sugiyama, Reiji … Show more authors
Yusuke Fujisawa, Sayaka Kikuchi, Fujino Kuba, Kosei Oishi, Soushi Murayama, Tomoya Sugiyama, Reiji Tokito, Hiroe Ueno, Shin-ichi Kashiwabara, Yasushi Yumura, and Yasuyuki Kurihara Hide authors
Reproductive Medicine and Biology 2024 [Reviewed]
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Single Work
陰陽のミトコンドリア 第１回ミトコンドリアは面白い
First Knowledge 第7号 ( 7 ) 3 - 6 2018.8
Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (bulletin of university, research institution) Publisher：株式会社 jast
Mechanism of mammalian germ cell development: toward new assisted reproductive technologies
Atsushi Suzuki, Yumiko SAGA
Strides of medicine 238 ( 13 ) 1221 - 1223 2011.9
Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (scientific journal) Joint Work
実験がうまくいく 蛍光・発光試薬の選び方と使い方、 「タンパク質検出法」
羊土社 2007.10
Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (other) Joint Work
ハツカネズミ亜種間雑種がもたらす染色体不安定性
アニメイト 2006
Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (scientific journal) Single Work
ハイブリドーマ細胞のスクリーニング方法
酒巻里菜、松葉隆雄、栗原靖之
Applicant：東ソー株式会社
Application no：特願2024-195425 Date applied：2025
Rights holder：酒巻里菜、松葉隆雄、栗原靖之
抗複数回膜貫通タンパク質モノクローナル抗体作製の革新的な新技術開拓とその普及
Grant number：2 4 K 0 8 1 6 2024.4 - 2027.3
Grant-in-Aid for Scientific Research(C)
Investigator(s)：栗原靖之
Authorship：Principal investigator Grant type：Competitive
本申請課題では、真核細胞の膜タンパク質(TM-タンパク質)に対する高品質モノクローナ
ル抗体（mAb）を、短期間に、効率よく、最少の手間で確実に取得する独自のハイブリド
ーマスクリーニング技術を開発する。これは、申請者が水溶性抗原に対するmAb取得の最適
解であるMIHS法をTM-タンパク質にも適用できるように改良し、その技術の普及を図るこ
とで我が国の研究開発力の向上に貢献することを目標にしている。
分子病理学的アプローチに基づく男性不妊症に関わるミトコンドリア電子伝達系制御機構
2021.4 - 2023.3
科学研究費補助金 Grant-in-Aid for Scientific Research(C)
Investigator(s)：栗原靖之
Grant type：Competitive
申請者を含めた多くの研究により、男性不妊症の約8割を占める造精障害ではミトコンドリア電子伝達系(Mt-ETC) の機能不全が起こっていることが示されている。この申請課題は、精子形成後期に特異的に発現するETC複合体Ⅳタンパク質(Coxfa4l3)の欠損（KO）マウスを、世界で初めての男性不妊症疾患モデル動物として利用して、男性不妊症が起こる分子メカニズムを分子病理学的アプローチで迫り、この治療法の開発に貢献する。
さらに、男性不妊症の有望な検査薬候補としてETC複合体Ⅳタンパク質を使った簡便な検査法の確立を目指す。これにより、多くの不妊に悩む人達に貢献したい。
体細胞超突然変異による高機能モノクローナル抗体を短期間に確実に取得する技術開発
2024.4 - 2025.10
オープンイノベーションプログ ラム NeyeS
Investigator(s)：栗原靖之
Authorship：Principal investigator
ミトコンドリアタンパク質を診断標的とする男性不妊症体外診断薬の開発
2021.4 - 2022.3
Japan Agency for Medical Research and Development 橋渡し研究戦略的推進プログラム
Investigator(s)：栗原靖之
Authorship：Principal investigator
迅速にミトコンドリア（関連）病をスクリーニング検査できる体外 診断薬開発
2020.4 - 2021.3
Japan Agency for Medical Research and Development
Investigator(s)：栗原靖之
Authorship：Principal investigator
様々なミトコンドリア病を迅速 。 簡便にスクリーニングできる抗体検査薬開発
2019.8 - 2020.3
Private Foundations
Authorship：Principal investigator
哺乳類精子の受精に欠かせない活発な精子運動性を支える新規エネルギー生産メカニズムの発見
大石航世, 時任怜史, 杉山智哉, 川村芽生, 三上小雪, 島田昌之, 渡部浩之, 手塚雅文, 浅田正嗣,村西由紀, 湯村 寧, 吉田 薫, 吉田 学, 栗原靖之
日本アンドロロジー学会 第44回学術大会＠神戸 2025.7
Event date： 2025.7
Language：Japanese Presentation type：Oral presentation (general)
Venue：神戸
MIHS法を用いた抗可溶性インターロイキン-2受容体抗体の開発
菊池 沙也香、黒山 浩之、横山 明彦、栗原 靖之
第3回日本抗体学会学術大会＠仙台
Event date： 2024.12
Language：English Presentation type：Oral presentation (general)
迅速・簡便なサンドイッチELISA抗体ペア・スクリーニング法の開発
青木 耕一郎, 川村 芽生, 久芳 藤乃, 栗原 靖之
第3回日本抗体学会学術大会
Event date： 2024.12
Language：Japanese Presentation type：Poster presentation
ミトコンドリア電子伝達系複合体IVのサブユニットタンパク質Coxfa4l3は精子運動性を高める
大石 航世、時任 怜史、杉山 智哉、吉田 学、吉田 薫、小出 剛、栗原 靖之
第47回日本分子生物学会
Event date： 2024.11
Language：English Presentation type：Oral presentation (general)
潜在的に電子伝達系活性を高める精子形成期の複合体Ⅳサブユニットタンパク質Coxfa4l3
時任 怜史、大石 航世、杉山 智哉、吉田 学、吉田 薫、今井 悠二、吉見 一人、小出 剛、栗原 靖之
第23回日本ミトコンドリア学会年会
Event date： 2024.11
Language：English Presentation type：Oral presentation (general)
新規モノクローナル抗体の作製
Reproduction mammalian genetics
2025 Seminar C in Chemistry Applications and Life Science
Graduate school of Engineering Science
2025 Functional Genome Science
Graduate school of Engineering Science
2025 Molecular Biology
College of Engineering Science
2025 Modern Biology 1
College of Engineering Science
2025 Cell Genetics
College of Engineering Science
医科学実験法Ⅰ、Ⅱ
Institution：横浜市立大学
国立大学法人 機器分析センター協議会
2020.4 会長
Committee type：Other
横浜市大と横浜国大、男性不妊治療法の選択に有用な新規バイオマーカー候補となるタンパク質を発見
日本経済新聞 2025.1
Author：Other
Une découverte d’anticorps qui suit le courant (流れに乗った抗体発見)
Nouvelles du monde 2024.3
Author：Other
横浜市立大、男性不妊の新規バイオマーカー候補となるタンパク質を発見
日経バイオテク
Author：Other
