Mechanism analysis of sulfobetaines' activity to prevent protein aggregation and application of sulfobetaines to unstable proteins

2011 Fiscal Year Final Research Report

• Project/Area Number: 21570108
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Gunma University
• Principal Investigator: WAKAMATSU Kaori 群馬大学, 大学院・工学研究科, 教授 (40222426)
• Co-Investigator(Renkei-kenkyūsha): SUGASE Kenji (財)サントリー生物有機科学研究所, 主席研究員 (00300822) ; NAMEKI Nobukazu 群馬大学, 大学院・工学研究科, 准教授 (80302959)
• Project Period (FY): 2009 – 2011
• Keywords: タンパク質 / 凝集防止 / 安定化 / スルホベタイン

Research Abstract

Aggregation of proteins brings about severe problems in drug development. Although sulfobetaines have been known to prevent protein aggregation, its mechanisms have been elusive. Here we found that sulfobetaines accelerate inter-conformer exchange rates of proteins and increase flexibility of loops. In addition, sulfobetaines were found to make proteins unstable depending on the identity of proteins and buffer conditions. Prevention of protein aggregation by sulfobetaines did not correlate with enhancement of thermostability of proteins. Sulfobetaines were found to be useful for protecting antibodies and for efficient purification of unstable proteins.

Research Products

• [Journal Article] Inhibitory effects of choline-O-sulfate on amyloid formation of human islet amyloid polypeptide (2012)
  • Author(s): M. Hagihara, A. Takei, T. Ishii, F. Hayashi, K. Kubota, K. Wakamatsu, N. Nameki
  • Journal Title: FEBS Open Bio Volume: 2 Pages: 20-25
  • Peer Reviewed
• [Journal Article] Additive effects of betaines on the fibrinogen cryogelation induced by low temperature (2011)
  • Author(s): Y. Toyama, K. Miyamoto, K. Kubota, K. Wakamatsu, N. Nobukazu, T. Saheki, M. Ochiai
  • Journal Title: Trans. Materials Res. Soc. J. Volume: 36 Pages: 393-396
  • Peer Reviewed
• [Journal Article] Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity (2011)
  • Author(s): Y. Konno, Y. Kobayashi, K. Takahashi, E. Takahashi, S. Sakae, M. Wakitani, K. Yamano, T. Suzawa1, K. Yano, T. Ohta, M. Koike, K. Wakamatsu, S. Hosoi
  • Journal Title: Cytotechmology
  • Peer Reviewed
• [Journal Article] Evaluation of the aggregation states of monoclonal antibodies by diverse and complementary methods (2011)
  • Author(s): T. Yoshino, T. Ishikawa, T. Ishihara, Y. Takeuchi, H. Yoshikawa, H. Yoshida, H. Yoshida, K. Wakamatsu
  • Journal Title: Biol. Pharm. Bull. Volume: 34 Pages: 1273-1278
  • Peer Reviewed
• [Journal Article] Enhancement of antibody production by the addition of Coenzyme-Q_<10> (2011)
  • Author(s): Y. Konno, M. Aoki, M. Takagishi, N. Sakai, M. Koike, K. Wakamatsu, S. Hosoi
  • Journal Title: Cytotechnology Volume: 63 Pages: 163-170
  • Peer Reviewed
• [Journal Article] Influence of pH on Heat-Induced Aggregation and Degradation of Therapeutic Monoclonal Antibodies (2010)
  • Author(s): T. Ishikawa, T. Ito, R. Endo, K. Nakagawa, E. Sawa, K. Wakamatsu
  • Journal Title: Biol. Pharm. Bull. Volume: 33 Pages: 1413-1417
  • Peer Reviewed
• [Journal Article] Relationship between human IgG structure and retention time in hydroxyapatite chromatography with sodium-chloride gradient elution (2010)
  • Author(s): T. Nakagawa, T. Ishihara, H. Yoshida, T. Yoshida, K. Wakamatsu, T. Kadoya
  • Journal Title: J. Sep. Sci. Volume: 33 Pages: 2045-2051
  • Peer Reviewed
• [Journal Article] Prevention of stirring-induced microparticle formation in monoclonal antibody solutions (2010)
  • Author(s): T. Ishikawa, N. Kobayashi, C. Osawa, E. Sawa, K. Wakamatsu
  • Journal Title: Biol. Pharm. Bull. Volume: 33 Pages: 1043-1046
  • Peer Reviewed
• [Journal Article] Relationship between human IgG structure and retention time in hydroxyapatite chromatography with sodium-phosphate gradient elution (2010)
  • Author(s): T. Nakagawa, T. Ishihara, H. Yoshida, T. Yoneya, K. Wakamatsu, T. Kadoya
  • Journal Title: J. Sep. Sci. Volume: 33 Pages: 37-45
  • Peer Reviewed
• [Journal Article] Binding site of C-reactive protein on M-ficolin (2009)
  • Author(s): M. Tanio, K. Wakamatsu, T. Kohno
  • Journal Title: Mol Immunol. Volume: 47 Pages: 215-221
  • Peer Reviewed
• [Journal Article] GDP-GTP exchange processes of G・i1 proten areaccelrated/decelerted depending on the type and the concentration of added detergents (2009)
  • Author(s): M. Kubota, T. Tanaka, T. Kohno, K. Wakamatsu
  • Journal Title: J. Biochem Volume: 146 Pages: 875-880
  • Peer Reviewed
• [Presentation] Stabilization of proteins by cholie-O-sulfate duing preparation and NMR measurement (2011)
  • Author(s): K. Wakamatsu, H. Wang, M. Takahashi, K. Nakazawa, M. Yoshizawa, T. Demura, K. Hosoda, S. Teraw, N. Nameki, K. Sugase
  • Organizer: The International Symposium on Nuclear Magnetic Resonance 2011
  • Place of Presentation: 横浜大さん橋ホール(神奈川県)
  • Year and Date: 2011-11-15
• [Presentation] 各種NDSBがタンパク質の凝集防止と安定化に及ぼす効果 (2011)
  • Author(s): 若松馨, 五十嵐麗樹, 金子敬輔, 高橋雅彦, 真井隆徳, 手塚福栄, 中沢和磨, 高橋玲羅, 王海梅, 石井毅, 細田和男, 寺脇慎一, 行木信一, 窪田健二
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都府)
  • Year and Date: 2011-09-21
• [Presentation] Elucidation of the mechanisms whereby NDSBs protect proteins from denaturation and aggregation (2011)
  • Author(s): M. Takahashi, K. Igarashi, K. Kaneko, E. Miura, H. Wang, T. Ishii, K. Kubota, K. Wakamatsu
  • Organizer: 第49回日本生物物理学会年会
  • Place of Presentation: 兵庫県立大学書写キャンパス(兵庫県)
  • Year and Date: 2011-09-16
• [Presentation] 蛋白質・ペプチドの溶液NMRへのスルホベタイン類の応用 (2010)
  • Author(s): 若松馨, 王海梅, 五十嵐麗樹, 神谷歩, 高橋雅彦, 手塚福栄, 向瓏, 石井毅, 細田和男, 行木信一, 菅瀬謙治
  • Organizer: 第49回NMR討論会
  • Place of Presentation: タワーホール船堀(東京都)
  • Year and Date: 2010-11-15
• [Presentation] NDSBか類蛋白質の & タイナミクスに及す効果 (2009)
  • Author(s): 若松馨, 王海梅, 松井文香, 神谷歩, 加藤瀬里, 行木信一, 菅瀬謙治
  • Organizer: 第48回NMR討論会
  • Place of Presentation: 九州大学(福岡県)
  • Year and Date: 2009-11-10
• [Remarks]
  • URL: http://wakamatsu-lab.dept.eng.gunma-u.ac.jp/Wakamatsu-lab/Tob.html