The effects of tween 20 and sucrose on the stability of anti-L-selectin during lyophilization and reconstitution
The effects of tween 20 and sucrose on the stability of anti-L-selectin during lyophilization and reconstitution
复制标题
DOI：
10.1002/jps.1098
复制
发表时间：
2001-10-01
影响因子：
3.8
通讯作者：
Carpenter, JF
中科院分区：
文献类型：
--
作者：
Jones, LS;Randolph, TW;Carpenter, JF
我们选择了抗L-选择素抗体作为模型蛋白，以研究蔗糖和/或吐温20对冻干和重构过程中蛋白质稳定性的影响。在蔗糖（1或0.125%）存在下，天然抗L-选择素二级结构在冻干过程中基本保留。然而，在不含蔗糖的情况下制备的冻干蛋白质粉末的复溶期间蛋白质的聚集不会因复溶介质中存在蔗糖而减少。通过用蔗糖冷冻干燥蛋白质并用0.1%吐温20溶液复溶，完全抑制复溶时的聚集体形成。吐温20（0.1%）还部分抑制冻干过程中天然抗L-选择素二级结构的损失。然而，复溶后，用吐温20冻干的制剂含有最高水平的聚集体。仅在复溶溶液中存在吐温似乎抑制了从二聚体向更高级低聚物的转变。研究了吐温20效应的潜在机制。然而，没有证据表明抗L-选择素构象的热力学稳定性（例如，通过Tween 20结合）。(C)2001 Wiley-Liss Inc.和美国制药协会。
We have chosen an anti-L-selectin antibody as a model protein to investigate the effects of sucrose and/or Tween 20 on protein stability during lyophilization and reconstitution. Native anti-L-selectin secondary structure is substantially retained during lyophilization in the presence of sucrose (1 or 0.125%). However, aggregation of the protein during reconstitution of lyophilized protein powders prepared without sucrose is not reduced by the presence of sucrose in the reconstitution medium. Aggregate formation upon reconstitution is completely inhibited by freeze drying the protein with sucrose and reconstituting with a 0.1% Tween 20 solution. Tween 20 (0.1%) also partially inhibits loss of native anti-L-selectin secondary structure during lyophilization. However, upon reconstitution the formulations lyophilized with Tween 20 contain the highest levels of aggregates. The presence of Tween in only the reconstitution solution appears to inhibit the transition from dimers to higher order oligomers. Potential mechanism(s) for the Tween 20 effects were investigated. However, no evidence of thermodynamic stabilization of anti-L-selectin conformation (e.g., by Tween 20 binding) could be detected. (C) 2001 Wiley-Liss Inc. and the American Pharmaceutical Association.