Impact of Freezing on pH of Buffered Solutions and Consequences for Monoclonal Antibody Aggregation
Impact of Freezing on pH of Buffered Solutions and Consequences for Monoclonal Antibody Aggregation
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DOI：
10.1002/btpr.377
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发表时间：
2010-05-01
影响因子：
2.9
通讯作者：
Singh, Satish K.
中科院分区：
文献类型：
--
作者：
Kolhe, Parag;Amend, Elizabeth;Singh, Satish K.
生物药物的大规模冷冻是一项重要的单元操作，它可以使材料具有制造灵活性和更长的使用周期。生物在冷冻溶液中的稳定性与许多问题有关，包括潜在的不稳定的pH变化。PH的变化是由pK(A)S的温度变化、溶解度限制、共晶结晶和低温浓缩引起的。大多数常见蛋白质配方缓冲液在冷冻状态下的pH变化尚未得到系统测量。在其他缓冲液中，组氨酸盐酸盐、乙酸钠、组氨酸醋酸酯、柠檬酸盐和琥珀酸在+25到-30℃的温度范围内观察到小于I的pH单位变化(增加)，而Tris-盐酸盐的pH单位增加类似于1.2 pH单位的增加。一般来说，一旦冷却到0℃以下，所有这些缓冲液的pH都会稳步上升。在添加了海藻糖的组氨酸缓冲液中配制的IgG2单抗表现出与缓冲液本身相同的pH行为。这种抗体在不同的配方中经历冷冻/解冻循环，代表了一个反映实际情况的宽过程(相变)时间范围。对反复冻融循环后的可溶性聚集体的测量表明，在这种情况下，pH的变化不是聚集体形成的一个因素，而是由是否存在非结晶的冷冻保护辅料所决定。在没有冷冻保护剂的情况下，较长的相变时间会导致较高的聚集性。(C)2009年美国化学工程师协会生物技术研究所。计划，26：727-733,2010
Freezing of biologic drug substance at large scale is an important unit operation that enables manufacturing flexibility and increased use-period for the material. Stability of the biologic in frozen solutions is associated with a number of issues including potentially destabilizing pH changes. The pH changes arise from temperature-associated change in the pK(a)s, solubility limitations, eutectic crystallization, and cryoconcentration. The pH changes for most of the common protein formulation buffers in the frozen state have not been systematically measured. Sodium phosphate buffer, a well-studied system, shows the greatest change in pH when going from +25 to -30 degrees C. Among the other buffers, histidine hydrochloride, sodium acetate, histidine acetate, citrate, and succinate, less than I pH unit change (increase) was observed over the temperature range from +25 to -30 degrees C, whereas Tris-hydrochloride had an similar to 1.2 pH unit increase. In general, a steady increase in pH was observed for all these buffers once cooled below 0 degrees C. A formulated IgG2 monoclonal antibody in histidine buffer with added trehalose showed the same pH behavior as the buffer itself This antibody in various formulations was subject to freeze/thaw cycling representing a wide process (phase transition) time range, reflective of practical situations. Measurement of soluble aggregates after repeated freeze-thaw cycles shows that the change in pH was not a factor for aggregate formation in this case, which instead is governed by the presence or absence of noncrystallizing cryoprotective excipients. In the absence of a cryoprotectant, longer phase transition times lead to higher aggregation. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 26: 727-733, 2010