Research activity information

• Tracking Particle‐Encapsulated DNA Across the Anion‐Exchange Chromatography Fractions of Recombinant Adeno‐Associated Virus Using Droplet Digital PCR and High‐Throughput Sequencing

  Noriko Hashiba, Yuzhe Yuan, Emi Itou‐Kudo, Kyoko Masumi‐Koizumi, Keisuke Yusa, Kazuhisa Uchida

  Jun. 2025, Biotechnology Journal

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Encapsulation of a Single-Stranded Form of DNA Impurities into the Capsid of a Recombinant Adeno-Associated Virus

  Kazuhisa Uchida, Emi Ito-Kudo, Kiyoko Higashiyama, Kyoko Masumi-Koizumi, Keisuke Yusa, Yuzhe Yuan

  May 2025, Human Gene Therapy

  Scientific journal


• Database Chemistry for Genomics-Based Safety and Quality Evaluation of Biologics

  Norichika Ogata, Tomoko Matsuda, Aoi Hosaka, Akihisa Shina, Noriko Hashiba, Kazuhisa Uchida, Yoshinori Kawabe, Masamichi Kamihira, Takanori Yamaoka, Hiromu Kurita, Noriko Yamano-Adachi, Takeshi Omasa

  Feb. 2025


• Concise Analysis of Single-Stranded DNA of Recombinant Adeno-Associated Virus By Automated Electrophoresis System

  Yuzhe Yuan, Kiyoko Higashiyama, Noriko Hashiba, Kyoko Masumi-Koizumi, Keisuke Yusa, Kazuhisa Uchida

  Feb. 2024, Human Gene Therapy

  Scientific journal


• Evaluation of next-generation sequencing performance for in vitro detection of viruses in biological products

  Takamasa Hirai, Kiyoko Kataoka, Yuzhe Yuan, Keisuke Yusa, Yoji Sato, Kazuhisa Uchida, Ken Kono

  Feb. 2024, Biologicals

  Scientific journal


• Applications and Limitations of Equilibrium Density Gradient Analytical Ultracentrifugation for the Quantitative Characterization of Adeno-Associated Virus Vectors.

  Kiichi Hirohata, Yuki Yamaguchi, Takahiro Maruno, Risa Shibuya, Tetsuo Torisu, Takayuki Onishi, Hideto Chono, Junichi Mineno, Yuan Yuzhe, Kiyoko Higashiyama, Kyoko Masumi-Koizumi, Kazuhisa Uchida, Takenori Yamamoto, Eriko Uchida, Takashi Okada, Susumu Uchiyama

  Adeno-associated virus (AAV) vectors are produced as a mixture of the desired particle (full particle, FP), which is filled with the designed DNA, product-related impurities such as particle without DNA (empty particle, EP), and aggregates. Cesium chloride or iodixanol equilibrium density gradient ultracentrifugation (DGE-UC) has been used for the purification of AAV vectors. DGE-UC can separate FP from impurities based on the difference in their buoyant densities. Here, we report the applications and limitations of equilibrium density gradient analytical ultracentrifugation (DGE-AUC) using a modern AUC instrument that employs DGE-UC principles for the characterization and quantitation of AAV vectors. We evaluated the quantitative ability of DGE-AUC in comparison with sedimentation velocity AUC (SV-AUC) or band sedimentation AUC (BS-AUC) using AAVs with different DNA lengths and different serotypes. DGE-AUC enabled the accurate quantification of the ratio of FP to EP when the AAV vector primarily contains these particles. Furthermore, we developed a new workflow to identify the components of separated peaks in addition to FP and EP. Ultraviolet absorption spectra obtained by multiwavelength detection can also support peak assignment following component identification. DGE-AUC experiments for AAV vectors have limitations with regard to minor components with low absorption at the detected wavelength or those with a density similar to that of major components of AAV vectors. DGE-AUC is the only analytical method that can evaluate particle density heterogeneity; therefore, SV-AUC or BS-AUC and DGE-AUC are complementary methods for reliable assessment of the purity of AAV vectors.

  Jan. 2024, Analytical chemistry, English, International magazine

  [Refereed]

  Scientific journal


• Association between adeno‐associated virus genomic titers and intracellular plasmid levels

  Kyoko Masumi‐Koizumi, Yuzhe Yuan, Kiyoko Higashiyama, Keisuke Yusa, Kazuhisa Uchida

  Jan. 2024, Engineering Reports

  Scientific journal


• Novel Approach for Obtaining Variable Domain of New Antigen Receptor with Different Physicochemical Properties from Japanese Topeshark (Hemitriakis japanica)

  Tomofumi Nakada-Masuta, Hiroyuki Takeda, Kazuhisa Uchida

  Oct. 2023, Marine Drugs

  Scientific journal


• Novel Approach for Obtaining VNAR with Different Physicochemical Properties from Japanese Topeshark (Hemitriakis japanica)

  Tomofumi Nakada-Masuta, Hiroyuki Takeda, Kazuhisa Uchida

  Oct. 2023


• Association between adeno‐associated virus genomic titers and intracellular plasmid levels

  Kyoko Masumi-Koizumi, yuzhe yuan, Kiyoko Higashiyama, Keisuke Yusa, KAZUHISA UCHIDA

  Wiley, Jun. 2023, Engineering Reports, English

  Scientific journal


• Quantitation of Residual Host Cell DNA in Recombinant Adeno-Associated Virus Using Droplet Digital Polymerase Chain Reaction

  Kiyoko Higashiyama, Yuzhe Yuan, Noriko Hashiba, Kyoko Masumi-Koizumi, Keisuke Yusa, Kazuhisa Uchida

  Jun. 2023, Human Gene Therapy

  Scientific journal


• Quantitation of Residual Host Cell DNA in Recombinant Adeno-Associated Virus Using Droplet Digital Polymerase Chain Reaction

  Kiyoko Higashiyama, Yuzhe Yuan, Noriko Hashiba, Kyoko Masumi-Koizumi, Keisuke Yusa, KAZUHISA UCHIDA

  Mary Ann Liebert Inc, Jun. 2023, Human Gene Therapy, 34(11-12) (11-12), 578 - 585, English

  Scientific journal


• Investigating critical thermal parameters for pre-analytical preparation of adeno-associated virus vector genome titration by droplet digital polymerase chain reaction

  Shunsuke SAITO, Akihiko KONDO, Kazuhisa UCHIDA

  {AMED} {iD}3 Catalyst Unit, 2023, Translational and Regulatory Sciences, English

  Scientific journal


• Practice of Human Resource Development Related to Biopharmaceutical Manufacturing and Quality Control and the International Contribution as Centers of Excellence of Asia-Pacific Economic Cooperation-Life Sciences Innovation Forum-Regulatory Harmonization Steering Committee

  Kazuhisa Uchida

  Pharmaceutical Society of Japan, Jul. 2022, YAKUGAKU ZASSHI, 142(7) (7), 749 - 753, English

  Scientific journal


• Infectivity assessment of porcine endogenous retrovirus using high-throughput sequencing technologies

  Kono, K., Kataoka, K., Yuan, Y., Yusa, K., Uchida, K., Sato, Y.

  2021, Biologicals, 71

  Scientific journal


• Viral safety testing for biopharmaceuticals: Current and future prospects

  Keisuke YUSA, Yuzhe YUSA, Kazuhisa UCHIDA

  {AMED} {iD}3 Catalyst Unit, 2020, Translational and Regulatory Sciences, 2(3) (3), 94 - 99, English

  Scientific journal


• A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing technologies

  Kono, K., Kataoka, K., Yuan, Y., Yusa, K., Uchida, K., Sato, Y.

  2020, Scientific Reports, 10(1) (1)

  Scientific journal


• A chromatographic approach for elevating the antibody-dependent cellular cytotoxicity of antibody composites

  Tojo, S., Okazaki, A., Wakitani, M., Shinkawa, T., Uchida, K., Suzawa, T.

  2009, Biological and Pharmaceutical Bulletin, 32(9) (9)

  Scientific journal


• Glycoform-dependent conformational alteration of the Fc region of human immunoglobulin G1 as revealed by NMR spectroscopy

  Yamaguchi, Y., Nishimura, M., Nagano, M., Yagi, H., Sasakawa, H., Uchida, K., Shitara, K., Kato, K.

  2006, Biochimica et Biophysica Acta - General Subjects, 1760(4) (4)

  Scientific journal


• Enhanced Fc-dependent cellular cytotoxicity of Fc fusion proteins derived from TNF receptor II and LFA-3 by fucose removal from asn-linked oligosaccharides

  Shoji-Hosaka, E., Kobayashi, Y., Wakitani, M., Uchida, K., Niwa, R., Nakamura, K., Shitara, K.

  2006, Journal of Biochemistry, 140(6) (6)

  Scientific journal


• Fucose removal from complex-type oligosaccharide enhances the antibody-dependent cellular cytotoxicity of single-gene-encoded bispecific antibody comprising of two single-chain antibodies linked to the antibody constant region

  Natsume, A., Wakitani, M., Yamane-Ohnuki, N., Shoji-Hosaka, E., Niwa, R., Uchida, K., Satoh, M., Shitara, K.

  2006, Journal of Biochemistry, 140(3) (3)

  Scientific journal


• Fucose removal from complex-type oligosaccharide enhances the antibody-dependent cellular cytotoxicity of single-gene-encoded antibody comprising a single-chain antibody linked the antibody constant region

  Natsume, A., Wakitani, M., Yamane-Ohnuki, N., Shoji-Hosaka, E., Niwa, R., Uchida, K., Satoh, M., Shitara, K.

  2005, Journal of Immunological Methods, 306(1-2) (1-2)

  Scientific journal


• IgG subclass-independent improvement of antibody-dependent cellular cytotoxicity by fucose removal from Asn²⁹⁷-linked oligosaccharides

  Niwa, R., Natsume, A., Uehara, A., Wakitani, M., Iida, S., Uchida, K., Satoh, M., Shitara, K.

  2005, Journal of Immunological Methods, 306(1-2) (1-2)

  Scientific journal


• Establishment of FUT8 knockout Chinese hamster ovary cells: An ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity

  Yamane-Ohnuki, N., Kinoshita, S., Inoue-Urakubo, M., Kusunoki, M., Iida, S., Nakano, R., Wakitani, M., Niwa, R., Sakurada, M., Uchida, K., Shitara, K., Satoh, M.

  2004, Biotechnology and Bioengineering, 87(5) (5)

  Scientific journal


• Engineering Chinese hamster ovary cells to maximize effector function of produced antibodies using FUT8 siRNA

  Mori, K., Kuni-Kamochi, R., Yamane-Ohnuki, N., Wakitani, M., Yamano, K., Imai, H., Kanda, Y., Niwa, R., Iida, S., Uchida, K., Shitara, K., Satoh, M.

  2004, Biotechnology and Bioengineering, 88(7) (7)

  Scientific journal


• Fucose Depletion from Human IgG1 Oligosaccharide Enhances Binding Enthalpy and Association Rate between IgG1 and FcγRIIIa

  Okazaki, A., Shoji-Hosaka, E., Nakamura, K., Wakitani, M., Uchida, K., Kakita, S., Tsumoto, K., Kumagai, I., Shitara, K.

  2004, Journal of Molecular Biology, 336(5) (5)

  Scientific journal


• The absence of fucose but not the presence of galactose or bisecting N-acetylglucosamine of human IgG1 complex-type oligosaccharides shows the critical role of enhancing antibody-dependent cellular cytotoxicity

  Shinkawa, T., Nakamura, K., Yamane, N., Shoji-Hosaka, E., Kanda, Y., Sakurada, M., Uchida, K., Anazawa, H., Satoh, M., Yamasaki, M., Hanai, N., Shitara, K.

  2003, Journal of Biological Chemistry, 278(5) (5)

  Scientific journal


• Capillary column high-performance liquid chromatographic-electrospray ionization triple-stage quadrupole mass spectrometric analysis of proteins separated by two-dimensional polyacrylamide gel electrophoresis application to cerebellar protein mapping

  Nakayama, H., Uchida, K., Shinkai, F., Shinoda, T., Okuyama, T., Seta, K., Isobe, T.

  1996, Journal of Chromatography A, 730(1-2) (1-2)

  Scientific journal


• Nuclear localization of p70-like protein in neurons of the superior colliculus and substantia nigra of the gerbil

  Seto-Ohshima, A., Karasawa, N., Nagatsu, I., Uchida, K., Isobe, T., Imai, S., Ozono, S., Onozuka, M.

  1995, Acta Histochemica et Cytochemica, 28(5) (5)

  Scientific journal


• Similarity of ribosomal proteins studied by two-dimensional coelectrophoresis for identification of gram-positive bacteria

  Ochiai, K., Uchida, K., Kawamoto, I.

  1993, International Journal of Systematic Bacteriology, 43(1) (1)

  Scientific journal


• Automated two-dimensional liquid chromatographic system for mapping proteins in highly complex mixtures

  Isobe, T., Uchida, K., Taoka, M., Shinkai, F., Manabe, T., Okuyama, T.

  1991, Journal of Chromatography A, 588(1-2) (1-2)

  Scientific journal


• Molecular cloning and characterization of a ras p21-like GTP-binding protein (24KG) from rat liver

  Sakurada, K., Uchida, K., Yamaguchi, K., Aisaka, K., Ito, S., Ohmori, T., Takeyama, Y., Ueda, T., Hori, Y., Ohyanagi, H., Saitoh, Y., Kaibuchi, K., Takai, Y.

  1991, Biochemical and Biophysical Research Communications, 177(3) (3)

  Scientific journal

• バイオ医薬品の製造・開発と規制上の課題

  UCHIDA KAZUHISA

  バイオ医薬品の製造・開発と規制上の課題の列挙と解決法の提示

  レギュラトリーサイエンス学会, May 2017, レギュラトリーサイエンス学会誌, 7(2) (2), 121 - 129, Japanese

  [Refereed]

  Introduction scientific journal

• バイオ医薬品の品質管理等に関わる人材育成プログラムの開発

  内田 和久

  日本医療研究開発機構, 創薬基盤推進研究事業, 2015, Principal investigator

  Competitive research funding