Hiruta, Yuki

写真a

Affiliation

Faculty of Science and Technology, Department of Applied Chemistry (Yagami)

Position

Associate Professor
Page Top ▲

Career 【 Display / hide 】

  • 2010.04
    -
    2011.03

    Keio University, Graduate School of Science and Technology, Assistant Professor

  • 2011.04
    -
    2013.03

    Japan Society for the Promition of Science, Research Fellowship for Young Scientists (DC2)

  • 2013.04
    -
    2017.03

    Keio University, Faculty of Pharmacy, Assistant Professor

  • 2017.04
    -
    2023.03

    Keio University, Faculty of Science and Technology, Senior Assistant Professor

  • 2023.04
    -
    Present

    Keio University, Faculty of Science and Technology, Associate Professor

display all >>

Page Top ▲

Academic Background 【 Display / hide 】

  • 2004.04
    -
    2008.03

    Keio University, Faculty of Science and Technology, Department of Applied Chemistry

    University, Graduated

  • 2008.04
    -
    2010.03

    Keio University, Graduate School of Science and Technology, School of Integrated Design Engineering

    Graduate School, Completed, Master's course

  • 2010.04
    -
    2013.03

    Keio University, Graduate School of Science and Technology, School of Integrated Design Engineering

    Graduate School, Completed, Doctoral course

Page Top ▲

Academic Degrees 【 Display / hide 】

  • 博士（工学）, 慶應義塾大学理工学研究科総合デザイン工学専攻, Dissertation

Page Top ▲
 

Books 【 Display / hide 】

  • Development of Near-Infrared Fluorescent Mg<sup>2+</sup> Probe and Application to Multicolor Imaging of Intracellular Signals

    Shindo Y., Ikeda Y., Hiruta Y., Citterio D., Oka K., Methods in Molecular Biology, 2021

    　View Summary

    Recent extensive studies revealed that the intracellular concentration of magnesium ions (Mg2+) is one of the important factors to regulate cellular functions. To evaluate the impact of Mg2+ concentration changes on intracellular signals or events, simultaneous imaging of Mg2+ with those phenomena is a powerful technique. The present protocol describes the synthesis and evaluation of near-infrared (NIR) fluorescent Mg2+-selective probes, named KMG-500 series, and the application to simultaneous imaging of the corresponding intracellular signal transductions and molecular events. The present protocol for multicolor imaging using fluorescent probes in the NIR and visible ranges is highly useful to reveal how multiple molecular events are correlated each other in each single cell.

Page Top ▲

Papers 【 Display / hide 】

  • Current advances in separation chemistry for antibody purification and analysis

    Deol S., Matsuda Y., Hiruta Y.

    Analytical Sciences 41 ( 5 ) 653 - 666 2025.05

    Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted,  ISSN  09106340

    　View Summary

    Monoclonal antibodies (mAbs) have become essential in modern therapeutics, offering high specificity and efficacy in treating diseases such as cancer, autoimmune disorders, and infectious diseases. With the increasing demand for mAbs, robust analytical and purification technologies are critical to ensuring their safety, efficacy, and consistency. This review highlights recent advances in the application of HPLC for both mAb analysis and purification. In the analytical domain, techniques such as size exclusion chromatography (SEC), hydrophobic interaction chromatography (HIC), ion exchange chromatography (IEX), and reversed-phase chromatography (RPLC) are explored for their role in evaluating mAb structural attributes and post-translational modifications (PTMs). These methods are indispensable for ensuring stability and functionality while meeting stringent regulatory requirements. In the context of purification, Protein A affinity chromatography remains the gold standard for initial capture of mAb; however, challenges such as high cost and harsh elution conditions have prompted the development of alternative purification technologies. IEX, HIC, multimodal, and membrane chromatography are also critical in achieving high-purity mAb products through multistep workflows, including intermediate purification and polishing stages. This review emphasizes the central role of HPLC in addressing the complex challenges of mAb manufacturing and characterization. By integrating established and emerging chromatographic techniques, it provides insights into the future directions of therapeutic antibody development.

  • Bioluminescence readout lateral flow immunoassay using nanobody targeting aflatoxin B1

    Takahashi, S; Hiruta, Y; Citterio, D

    ANALYST 150 ( 8 ) 1563 - 1570 2025.04

    Research paper (scientific journal), Joint Work, Accepted,  ISSN  0003-2654

    　View Summary

    Multiple signal detection methods are known for lateral flow immunoassays (LFIAs), with colorimetric approaches dominating the field. However, their limited sensitivity is a remaining challenge. Fluorescence-based signaling is regarded as a more sensitive method, but it comes at the cost of partial sacrifice of the user-friendliness of LFIAs due to the requirement of an excitation light source. In this context, bioluminescence providing an inherently high signal to noise ratio without the need of excitation light could be an attractive alternative. But only a few studies have demonstrated the application of bioluminescence signaling in LFIAs. This work aimed at the development of a simple bioluminescence-based LFIA for the detection of aflatoxin B1 (AFB1), used as a model target in a competitive LFIA format. Signal transduction was achieved by nanobody-nanoluciferase (Nluc) fusion proteins. These small-sized recombinant heavy-chain-only antibodies derived from the camelidae family directly linked with the Nluc enzyme produce high intensity glow-type bioluminescence in combination with the furimazine substrate. LFIA devices consisting of a sample pad, nitrocellulose membrane and absorbent pad with AFB1-BSA conjugate deposited at the test line on the nitrocellulose membrane, achieved an LOD of 0.26 ng mL<sup>−1</sup> for aqueous AFB1 solutions pre-mixed with Nanobody-Nluc and bioluminescence emission observed on an imaging system. More user-friendly LFIA devices with integrated conjugate pad and pre-deposited Nanobody-Nluc provided clear AFB1 concentration-dependent bioluminescence signals with low background and enabled readout with a standard digital camera, resulting in an LOD of 1.12 ng mL<sup>−1</sup>. Finally, the LFIA strips have been applied in AFB1-spiked oat milk samples. The LOD of 4.09 ng mL<sup>−1</sup> achieved in the real sample matrix is well below the maximum allowable residual concentration of AFB1 in the U.S. (20 ng mL<sup>−1</sup>).

  • Origami Paper-Based Immunoassay Device with CRISPR/Cas12a Signal Amplification

    Suzuki, H; Tong, GD; Nath, P; Hiruta, Y; Citterio, D

    ACS SENSORS 10 ( 3 ) 1811 - 1821 2025.03

    Research paper (scientific journal), Joint Work, Accepted,  ISSN  2379-3694

    　View Summary

    In clinical diagnosis, the determination of target proteins at low concentration levels is generally performed by immunoassays, such as the enzyme-linked immunosorbent assay (ELISA), which is a time-consuming process. To date, paper-based ELISA platforms enabling faster and less expensive analysis have been developed, but their important issue for clinical applications is the limited sensitivity compared to conventional ELISA. To address this challenge, this paper introduces a simple, rapid, and highly sensitive detection method for non-nucleic acid targets achieved by integrating the CRISPR/Cas12a system into paper-based ELISA. An origami-type paper-based device enabling simple assay operation has been designed, and the detection of targets on the paper substrates is based on observing the fluorescence signal induced by the CRISPR/Cas12a enzyme cleaving a probe single-stranded DNA (ssDNA) labeled with fluorophore and quencher (FQ reporter). To enhance sensitivity, antibodies labeled with a network of multiple DNA activating the CRISPR/Cas12a enzyme have been utilized as detection antibodies. As a result, the developed device successfully boosted the detection sensitivity for both human IgG and the hepatitis B virus surface antigen (HBsAg). In particular, the limit of detection (LOD) for HBsAg was estimated to be 12 pg/mL, representing over 10-fold higher sensitivity compared with commercially available HBsAg ELISA kits (LOD: 200 pg/mL). In addition, the fluorescence response toward porcine whole blood samples containing different HBsAg concentrations was also confirmed by capturing images with a smartphone, followed by quantitative data analysis. These results demonstrate the potential applicability of the proposed platform for clinical tests at the point of care.

  • The latest developments of near-infrared fluorescent probes from NIR-I to NIR-II for bioimaging

    Kuronuma, Y; Watanabe, R; Hiruta, Y

    ANALYTICAL SCIENCES 41 ( 6 ) 737 - 757 2025.02

    Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted,  ISSN  0910-6340

    　View Summary

    Abstract: Near-infrared I (NIR-I: 650–950 nm) fluorescence imaging is a powerful tool for deep-tissue biological imaging, addressing the limitations of photon penetration depth in the visible-light region. Over the past decade, NIR imaging has extended to the near-infrared II (NIR-II: 1000–1700 nm) region, offering high-resolution and low background imaging by suppressing light scattering, and autofluorescence of tissues. Near-infrared fluorescent probes from NIR-I to NIR-II, with diverse functionalities, are increasingly utilized across biological fields to meet various detection needs and to explore physiological events in real time and spatial dimensions. This review discusses recent advancements in small-molecule NIR fluorescent dyes and probes, particularly those based on cyanine and rhodamine scaffolds, highlighting examples of their applications in bioimaging.

  • Ratiometric Imaging for Quantification of Elevated Ca<sup>2+</sup> in Neurons Using Synthetic Low-Affinity Fluorescent Probe

    Kuronuma Y., Shindo Y., Kumada R., Sakama A., Citterio D., Oka K., Hiruta Y.

    ACS Chemical Neuroscience 16 ( 4 ) 649 - 658 2025.02

    Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted,  ISSN  1948-7193

    　View Summary

    The availability of various calcium ion (Ca2+) fluorescent probes has contributed to revealing physiological events related to intracellular Ca2+. However, conventional probes face challenges for quantitatively and selectively visualizing high Ca2+ concentrations in cells induced by any stimuli, including biomolecules or electrical signal that disrupt Ca2+ homeostasis. In this report, we designed and synthesized a low-affinity ratiometric Ca2+ probe, KLCA-Fura, utilizing o-aminophenol-N,N-diacetate-O-methylene-methylphosphinate (APDAP) as a ligand, for which we recently demonstrated the suitability as a new low-affinity ligand for Ca2+. KLCA-Fura showed a blue shift in excitation wavelength with increasing Ca2+ concentration based on the intramolecular charge transfer (ICT). Its affinity for Ca2+ is lower than commercially available conventional Ca2+ probes. Furthermore, the selectivity for Ca2+ and the fluorescence intensity were considered sufficient to accurately detect Ca2+. The corresponding acetoxymethyl ester, KLCA-FuraAM, was synthesized for intracellular imaging and applied to Ca2+ quantification in neurons. KLCA-FuraAM enabled quantitative ratiometric monitoring of the two-step Ca2+ concentration increase induced by glutamate stimulation. While this two-step response was not clearly observed with a commercially available low-affinity ratiometric Ca2+ probe, Fura-FF, KLCA-FuraAM has demonstrated the potential to quantitatively visualize the behavior of high Ca2+ concentrations. The ratiometric low-affinity Ca2+ probe, KLCA-Fura, is expected to be a powerful tool for discovering new functions of Ca2+ in neurons.

display all >>

Page Top ▲

Papers, etc., Registered in KOARA 【 Display / hide 】

display all >>

Page Top ▲

Presentations 【 Display / hide 】

  • Development of column packing materials using functional polymers and application to pharmaceutical analysis

    The 35th Conference of Chromatographic Sciences, 

    2024.11
    , 

    Oral presentation (invited, special)

  • ペプチドミメティック分子設計によるpH応答性高分子の開発

    蛭田 勇樹

    第73回高分子討論会, 

    2024.09
    , 

    Oral presentation (general)

  • スピロ環化平衡を利用したpH応答性近赤外蛍光シアニンプローブ

    蛭田 勇樹

    日本分析化学会第73年会, 

    2024.09
    , 

    Oral presentation (general)

  • Cation/anion-exchange mode switching chromatography utilizing pH-responsive polymer for pharmaceutical separations

    Yuki Hiruta

    HPLC 2024, 

    2024.07
    , 

    Poster presentation

  • Evaluation of hydrophilic brush copolymers on biodistribution for tumor-targeted carriers of diagnostic drugs

    Yuki Hiruta

    第40回日本DDS学会学術集会, 

    2024.07
    , 

    Oral presentation (general)

display all >>

Page Top ▲

Research Projects of Competitive Funds, etc. 【 Display / hide 】

  • がんバイオマーカー応答性高分子を基盤とした光セラノスティクスプラットフォーム構築

    2023.04
    -
    2026.03

    文部科学省, 日本学術振興会 科学研究費助成事業 国際共同研究加速基金(国際共同研究強化(A)), Principal investigator

  • 生命科学研究を拓く生物発光技術の開発

    2023.04
    -
    2024.03

    文部科学省, 日本学術振興会 科学研究費助成事業 基盤研究(B), Coinvestigator(s)

  • がんバイオマーカー応答性高分子設計を戦略とした光セラノスティクス薬剤の開発

    2021.04
    -
    2024.03

    MEXT,JSPS, Grant-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Principal investigator

  • 時空間トランススケールイメージングを可能にする超分子ケージドルシフェリンの開発

    2021.04
    -
    2023.03

    MEXT,JSPS, Grant-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas, Principal investigator

  • Development of polymer caged luciferin enabling spatiotemporal trans-scale imaging

    2019.04
    -
    2021.03

    MEXT,JSPS, Grant-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas, Principal investigator

display all >>

Page Top ▲

Awards 【 Display / hide 】

  • Award for Encouragement of Research in Chromatographic Sciences

    2024.11, The Society for Chromatographic Sciences, Development of column packing materials using functional polymers and application to pharmaceutical analysis

    Type of Award： Award from Japanese society, conference, symposium, etc.

  • Best Poster Award HPLC 2022, San Diego

    2022.06

    Type of Award： Award from international society, conference, symposium, etc.

  • Outstanding Reviewer for journal of Materials Chemistry B

    2021.03

    Type of Award： Award from publisher, newspaper, foundation, etc.

  • Award for Encouragement of Research in Analytical Chemistry

    2020.08, The Japan Society of Analytical Chemistry, Development and application of bioimaging probes based on precision molecular design

    Type of Award： Award from Japanese society, conference, symposium, etc.

  • Award for Encouragement of Research in Polymer Science

    Development of stimuli-responsive polymer materials for application to medical analysis, 2020.05, 公益社団法人高分子学会, 医療分析への応用を志向した刺激応答性高分子材料の開発

    Type of Award： Award from Japanese society, conference, symposium, etc.

display all >>

Page Top ▲
 

Courses Taught 【 Display / hide 】

  • SEMINAR IN APPLIED CHEMISTRY

    2025

  • PRACTICAL INSTRUMENTAL ANALYSIS

    2025

  • NANO SCALE SCIENCE JOINT SEMINAR

    2025

  • MATERIAL DESIGN SCIENCE JOINT SEMINAR

    2025

  • LABORATORY IN SCIENCE

    2025

display all >>

Page Top ▲
 

Memberships in Academic Societies 【 Display / hide 】

  • クロマトグラフィー科学会会員, 

    2016.01
    -
    Present

  • 高分子学会会員, 

    2016.01
    -
    Present

  • 日本DDS学会会員, 

    2014.04
    -
    Present

  • 日本薬学会会員, 

    2014.01
    -
    Present

  • 日本化学会会員, 

    2010.01
    -
    Present

display all >>

Page Top ▲

Committee Experiences 【 Display / hide 】

  • 2024.11
    -
    Present

    生物発光化学発光研究会世話人, 生物発光化学発光研究会

  • 2024.03
    -
    Present

    Associate Editor in Analytical Sciences, The Japan Society for Analytical Chemistry

  • 2021.01
    -
    2022.12

    関東支部常任幹事 , 日本分析化学会

  • 2019.12
    -
    2021.11

    化学グランプリ・オリンピック委員会 グランプリ小委員会 委員, 公益社団法人日本化学会

  • 2018.04
    -
    Present

    代議員, 日本分析化学会

display all >>

Page Top ▲