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Effect of high DMSO concentration on albumin during freezing and vitrification

机译：高达DMSO浓度对冻融和玻璃化期间白蛋白的影响

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摘要

IR spectroscopy was used to probe the composition of the freeze concentrated liquid (FCL) during near equilibrium freezing of albumin-dimethyl sulfoxide (DMSO) solutions down to -70 degrees C. While cooling down to -32 degrees C, preferential entrapment of DMSO within ice occurred, which was attributed to formation of DMSO- water complexes. These complexes subsequently collapsed on cooling below -32 degrees C with albumin being preferentially entrapped in ice. Heterogeneity induced in the FCL as a result, was more pronounced in solutions containing lower solute concentrations and thus larger volume fractions of ice. Crystallization of DMSO was not observed on cooling below its eutectic temperature and the FCL vitrified instead at -58 degrees C, as confirmed by differential scanning calorimetry and low temperature X-ray crystallography. The increase in viscosity of the FCL upon glass transition prevented further compositional changes. Isothermal measurements confirmed that preferential exclusion of either species from ice occurred solely during advance of the ice interfaces and was hence governed by its relative diffusivity. Adverse effects of high DMSO concentrations on the protein were observed between -35 degrees C and -55 degrees C and devitrification during thawing proved to be even more detrimental in vitrified samples. Thawing rate had a significant effect on the extent of structural unfolding of the protein in vitrified samples, but not on the ones which were simply frozen. These results will be relevant to vitrification of solutions containing proteins, cells and small tissue systems as such as monocytes, oocytes, ova, embryos, zygotes, vein rings/segments and pancreatic islets.
机译：IR光谱法用于探测在近距离白蛋白 - 二甲基磺氧化物（DMSO）溶液的接近平衡至-70℃的近平均浓缩液体（FCL）的组合物。在冷却至-32摄氏度下，优先捕获DMSO内发生了冰，这归因于形成DMSO-水络合物。这些配合物随后在低于-32℃的冷却后塌陷，用白蛋白优先捕获在冰中。因此，在FCL中诱导的异质性在含有较低溶质浓度的溶液中更加明显，因此更大体积冰。通过差示扫描量热法和低温X射线晶体学证实，未观察到DMSO的结晶而不是在其共晶温度和-58℃下的vcl vitrizer代替。玻璃化转变对FCL的粘度的增加防止了进一步的组成变化。等温测量证实，在冰界面的前进期间，仅从冰中发生了从冰中出现的任何种类，因此通过其相对扩散性来治理。高DMSO浓度对蛋白质对蛋白质的不利影响在-35℃和-55℃之间观察到，在解冻期间的缺失被证明在玻璃化样品中是更有害的。解冻率对玻璃化样品中蛋白质的结构展开的程度具有显着影响，但不是简单地冷冻的结构展开的程度。这些结果将与含有蛋白质，细胞和小组织系统的溶液的玻璃化相关，例如单核细胞，卵母细胞，卵子，胚胎，Zygotes，静脉环/段和胰岛。

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  • 来源
    《RSC Advances》 |2017年第69期|共10页
  • 作者

    Jena Sampreeti; Aksan Alptekin;

  • 作者单位

    Univ Minnesota Dept Mech Engn Biostabilizat Lab 111 Church St SE Minneapolis MN 55455 USA;

    Univ Minnesota Dept Mech Engn Biostabilizat Lab 111 Church St SE Minneapolis MN 55455 USA;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
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