Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage

Hideto Hirahara, Yutaka Nagare, Alan Twomey, Kosaku Kurata, Takanobu Fukunaga, Alptekin Aksan, Hiroshi Takamatsu

研究成果: 会議への寄与タイプ論文

抄録

Stability and activities of biopharmaceuticals after thawing largely depend on ice-solute interaction in freezing process. The aims of the present study were to observe microscopic distribution of protein or sugar that was trapped in the ice after freezing of their aqueous solutions and to evaluate protein denaturation. A confocal Raman microscope (CRM) combined with a directional solidification stage (DSS) was developed for this purpose. Trehalose and bovine serum albumin solutions at a concentration of 5 to 20 wt % were respectively enclosed in a microchannel. They were solidified on the DSS at different degree of supercooling, and then scanned by the CRM along the microchannel. The Raman imaging successfully exhibited variety in distribution of ice and solutes: uniform dispersion of the organic matters and ice crystals at highly supercooled area because of higher freezing speed, formation of solute-accumulated spots between ice crystals at lower supercooled area due to rejection of solutes, and relatively pure ice region near the ice front due to full rejection of solutes. It depended on the solute and its concentration, the degree of supercooling, and the freezing speed. An attempt to evaluate the freeze-induced denaturation of protein was also made with analyzing the Raman spectrum of α-helix band relative to that of CH band. An increase in denaturation was observed with repetitive freezing, which demonstrated the feasibility of denaturation detection. It however suggested that using the other type of protein that is more sensitive to freezing is preferable to evaluate site-dependent denaturation in-situ.

元の言語英語
出版物ステータス出版済み - 1 1 2014
イベント15th International Heat Transfer Conference, IHTC 2014 - Kyoto, 日本
継続期間: 8 10 20148 15 2014

その他

その他15th International Heat Transfer Conference, IHTC 2014
日本
Kyoto
期間8/10/148/15/14

Fingerprint

albumins
Freezing
freezing
Ice
Solidification
Denaturation
solutes
biopolymer denaturation
ice
Microscopes
microscopes
Proteins
interactions
Supercooling
supercooling
microchannels
Microchannels
proteins
rejection
Crystals

All Science Journal Classification (ASJC) codes

  • Mechanical Engineering
  • Condensed Matter Physics

これを引用

Hirahara, H., Nagare, Y., Twomey, A., Kurata, K., Fukunaga, T., Aksan, A., & Takamatsu, H. (2014). Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage. 論文発表場所 15th International Heat Transfer Conference, IHTC 2014, Kyoto, 日本.

Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage. / Hirahara, Hideto; Nagare, Yutaka; Twomey, Alan; Kurata, Kosaku; Fukunaga, Takanobu; Aksan, Alptekin; Takamatsu, Hiroshi.

2014. 論文発表場所 15th International Heat Transfer Conference, IHTC 2014, Kyoto, 日本.

研究成果: 会議への寄与タイプ論文

Hirahara, H, Nagare, Y, Twomey, A, Kurata, K, Fukunaga, T, Aksan, A & Takamatsu, H 2014, 'Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage' 論文発表場所 15th International Heat Transfer Conference, IHTC 2014, Kyoto, 日本, 8/10/14 - 8/15/14, .
Hirahara H, Nagare Y, Twomey A, Kurata K, Fukunaga T, Aksan A その他. Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage. 2014. 論文発表場所 15th International Heat Transfer Conference, IHTC 2014, Kyoto, 日本.
Hirahara, Hideto ; Nagare, Yutaka ; Twomey, Alan ; Kurata, Kosaku ; Fukunaga, Takanobu ; Aksan, Alptekin ; Takamatsu, Hiroshi. / Observation of ice-solute interaction in freezing of trehalose and albumin solutions by using confocal Raman microscope equipped with directional solidification stage. 論文発表場所 15th International Heat Transfer Conference, IHTC 2014, Kyoto, 日本.
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abstract = "Stability and activities of biopharmaceuticals after thawing largely depend on ice-solute interaction in freezing process. The aims of the present study were to observe microscopic distribution of protein or sugar that was trapped in the ice after freezing of their aqueous solutions and to evaluate protein denaturation. A confocal Raman microscope (CRM) combined with a directional solidification stage (DSS) was developed for this purpose. Trehalose and bovine serum albumin solutions at a concentration of 5 to 20 wt {\%} were respectively enclosed in a microchannel. They were solidified on the DSS at different degree of supercooling, and then scanned by the CRM along the microchannel. The Raman imaging successfully exhibited variety in distribution of ice and solutes: uniform dispersion of the organic matters and ice crystals at highly supercooled area because of higher freezing speed, formation of solute-accumulated spots between ice crystals at lower supercooled area due to rejection of solutes, and relatively pure ice region near the ice front due to full rejection of solutes. It depended on the solute and its concentration, the degree of supercooling, and the freezing speed. An attempt to evaluate the freeze-induced denaturation of protein was also made with analyzing the Raman spectrum of α-helix band relative to that of CH band. An increase in denaturation was observed with repetitive freezing, which demonstrated the feasibility of denaturation detection. It however suggested that using the other type of protein that is more sensitive to freezing is preferable to evaluate site-dependent denaturation in-situ.",
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AU - Hirahara, Hideto

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AU - Kurata, Kosaku

AU - Fukunaga, Takanobu

AU - Aksan, Alptekin

AU - Takamatsu, Hiroshi

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