Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Biotechnol Bioeng. 1992 Oct 20;40(8):895-903. doi: 10.1002/bit.260400805.
Undesirable aggregation of aqueous insulin solutions remains a serious obstacle in the development of alternative methods of diabetes therapy. We investigated the fundamental nature of the aggregation mechanism and proposed stabilization strategies based on a mathematical model for the reaction scheme. Insulin aggregation kinetics in the presence of solid-liquid and air-liquid interfaces were monitored using UV spectroscopy and quasielastic light scattering (QELS). Experimental observations were consistent with our model of monomer denaturation at hydrophobic surfaces followed by the formation of stable intermediate species which facilitated subsequent macroaggregation. The model was used to predict qualitative trends in insulin aggregation behavior, to propose stabilization strategies, and to elucidate mechanisms of stabilization. In the absence of additives, insulin solutions aggregated completely (more than 95% of the soluble protein lost) within 24 h; with sugar-based nonionic detergents, no detectable loss occurred for more than 6 weeks.
水性胰岛素溶液的不良聚集仍然是开发糖尿病治疗替代方法的严重障碍。我们研究了聚集机制的基本性质，并基于反应方案的数学模型提出了稳定策略。使用紫外光谱法和准弹性光散射（QELS）监测固-液和气-液界面存在时的胰岛素聚集动力学。实验观察与我们的模型一致，即单体在疏水性表面变性，然后形成稳定的中间产物，促进随后的大聚集。该模型用于预测胰岛素聚集行为的定性趋势，提出稳定策略，并阐明稳定机制。在没有添加剂的情况下，胰岛素溶液在 24 小时内完全聚集（超过 95%的可溶性蛋白丢失）；使用基于糖的非离子型清洁剂，超过 6 周没有检测到明显的损失。
