Pharmaceutical Sciences, Research and Development, AbbVie Inc., 2525 Dupont Drive, Irvine, CA 92612, USA.
Center for Neutron Research, National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899, USA.
J Pharm Sci. 2023 Jan;112(1):76-82. doi: 10.1016/j.xphs.2022.08.017. Epub 2022 Aug 20.
Protein structural changes during freezing and subsequent thawing are of great importance to a variety of biopharmaceutical applications. In this work, we studied the influence of non-ionic surfactants (polysorbate 20 and poloxamer 188) on protein structural changes during freeze and thaw using lysozyme as a model protein. Small-angle neutron scattering was employed to characterize protein structures in both liquid and frozen solution states. The results show minimal impact of polysorbate 20 on lysozyme structures during freeze and thaw using practically relevant concentrations. Polysorbate 20 used at 0.04% (w/w) completely prevents freeze-induced aggregation of lysozyme. Poloxamer 188 seems to interact with lysozyme; when applied at high concentrations (10% w/w), such interaction prevents protein crowding or close packing typically associated with freeze concentration. Despite such interactions, lysozyme aggregation is observed with 10% (w/w) of poloxamer 188 during freezing, although the aggregation is reversed upon thawing.
蛋白质在冷冻和随后解冻过程中的结构变化对各种生物制药应用都非常重要。在这项工作中，我们以溶菌酶为模型蛋白，研究了非离子表面活性剂（聚山梨酯 20 和泊洛沙姆 188）在冷冻和解冻过程中对蛋白质结构变化的影响。小角中子散射被用来在液体和冷冻溶液状态下对蛋白质结构进行表征。结果表明，在实际相关浓度下，聚山梨酯 20 对溶菌酶结构在冷冻和解冻过程中的影响很小。0.04%（w/w）的聚山梨酯 20 可完全防止溶菌酶在冷冻诱导下聚集。泊洛沙姆 188 似乎与溶菌酶相互作用；当应用于高浓度（10%w/w）时，这种相互作用可防止通常与冷冻浓缩相关的蛋白质拥挤或紧密堆积。尽管存在这种相互作用，但在 10%（w/w）的泊洛沙姆 188 存在下仍观察到溶菌酶在冷冻过程中的聚集，尽管在解冻时聚集被逆转。
