Research output: Contribution to journal › Article › peer-review
Ice Recrystallization in a Solution of a Cryoprotector and Its Inhibition by a Protein : Synchrotron X-Ray Diffraction Study. / Zakharov, Boris; Fisyuk, Alexander; Fitch, Andy et al.
In: Journal of Pharmaceutical Sciences, Vol. 105, No. 7, 01.07.2016, p. 2129-2138.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Ice Recrystallization in a Solution of a Cryoprotector and Its Inhibition by a Protein
T2 - Synchrotron X-Ray Diffraction Study
AU - Zakharov, Boris
AU - Fisyuk, Alexander
AU - Fitch, Andy
AU - Watier, Yves
AU - Kostyuchenko, Anastasia
AU - Varshney, Dushyant
AU - Sztucki, Michael
AU - Boldyreva, Elena
AU - Shalaev, Evgenyi
PY - 2016/7/1
Y1 - 2016/7/1
N2 - Ice formation and recrystallization is a key phenomenon in freezing and freeze-drying of pharmaceuticals and biopharmaceuticals. In this investigation, high-resolution synchrotron X-ray diffraction is used to quantify the extent of disorder of ice crystals in binary aqueous solutions of a cryoprotectant (sorbitol) and a protein, bovine serum albumin. Ice crystals in more dilute (10 wt%) solutions have lower level of microstrain and larger crystal domain size than these in more concentrated (40 wt%) solutions. Warming the sorbitol–water mixtures from 100 to 228 K resulted in partial ice melting, with simultaneous reduction in the microstrain and increase in crystallite size, that is, recrystallization. In contrast to sorbitol solutions, ice crystals in the BSA solutions preserved both the microstrain and smaller crystallite size on partial melting, demonstrating that BSA inhibits ice recrystallization. The results are consistent with BSA partitioning into quasi-liquid layer on ice crystals but not with a direct protein–ice interaction and protein sorption on ice surface. The study shows for the first time that a common (i.e., not-antifreeze) protein can have a major impact on ice recrystallization and also presents synchrotron X-ray diffraction as a unique tool for quantification of crystallinity and disorder in frozen aqueous systems.
AB - Ice formation and recrystallization is a key phenomenon in freezing and freeze-drying of pharmaceuticals and biopharmaceuticals. In this investigation, high-resolution synchrotron X-ray diffraction is used to quantify the extent of disorder of ice crystals in binary aqueous solutions of a cryoprotectant (sorbitol) and a protein, bovine serum albumin. Ice crystals in more dilute (10 wt%) solutions have lower level of microstrain and larger crystal domain size than these in more concentrated (40 wt%) solutions. Warming the sorbitol–water mixtures from 100 to 228 K resulted in partial ice melting, with simultaneous reduction in the microstrain and increase in crystallite size, that is, recrystallization. In contrast to sorbitol solutions, ice crystals in the BSA solutions preserved both the microstrain and smaller crystallite size on partial melting, demonstrating that BSA inhibits ice recrystallization. The results are consistent with BSA partitioning into quasi-liquid layer on ice crystals but not with a direct protein–ice interaction and protein sorption on ice surface. The study shows for the first time that a common (i.e., not-antifreeze) protein can have a major impact on ice recrystallization and also presents synchrotron X-ray diffraction as a unique tool for quantification of crystallinity and disorder in frozen aqueous systems.
KW - calorimetry (DSC)
KW - crystal defects
KW - freeze drying/lyophilization
KW - phase transition
KW - proteins
KW - X-ray powder diffractometry
UR - http://www.scopus.com/inward/record.url?scp=84975842553&partnerID=8YFLogxK
U2 - 10.1016/j.xphs.2016.04.020
DO - 10.1016/j.xphs.2016.04.020
M3 - Article
C2 - 27287516
AN - SCOPUS:84975842553
VL - 105
SP - 2129
EP - 2138
JO - Journal of Pharmaceutical Sciences
JF - Journal of Pharmaceutical Sciences
SN - 0022-3549
IS - 7
ER -
ID: 25461836