FOURIER-TRANSFORM INFRARED SPECTROSCOPIC INVESTIGATION OF PROTEIN STABILITY IN THE LYOPHILIZED FORM
FOURIER-TRANSFORM INFRARED SPECTROSCOPIC INVESTIGATION OF PROTEIN STABILITY IN THE LYOPHILIZED FORM
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DOI：
10.1016/0167-4838(95)00156-o
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发表时间：
1995-11-15
期刊：
影响因子：
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通讯作者：
KLIBANOV, A
中科院分区：
文献类型：
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作者：
COSTANTINO, HR;GRIEBENOW, K;KLIBANOV, A
在除去水后，蛋白质经历了一个主要的，可逆的重排其二级结构，如FTIR光谱所揭示的。我们在此发现，对于重组人白蛋白（rHA），这种结构变化的程度并不显著依赖于冻干前水溶液的组成（蛋白质浓度、pH和赋形剂如葡聚糖或NaCl的存在）或脱水模式（冻干、喷雾干燥或旋转蒸发），即使这些因素深刻地影响rKA的固态稳定性以抵抗水分诱导的聚集。在所有情况下，rHA的α-螺旋含量从溶液中的58%下降到脱水状态下的25 - 35%，β-折叠含量从0上升到10 - 20%，无序结构从40%增加到50 - 60%。我们还研究了另一种模型蛋白，鸡蛋清溶菌酶，并证实，它也经历了一个显着的改变，在冻干后的二级结构。已经发现这种结构重组的程度对冻干前水溶液的pH从pH 1.9至5.1不敏感，即使水溶液中的热转变温度（T-m）在该范围内变化30 ℃。
Upon the removal of water, proteins undergo a major, reversible rearrangement of their secondary structure, as revealed by FTIR spectroscopy. We have found herein that for recombinant human albumin (rHA) the extent of this structural change does not depend significantly either on the composition of the aqueous solution prior to lyophilization (protein concentration, pH, and the presence of excipients such as dextran or NaCl) or on the mode of dehydration (lyophilization, spray drying, or rotary evaporation), even though these factors profoundly affect rKA's solid-state stability against moisture-induced aggregation. In all cases, the alpha-helix content of rHA drops from 58% in solution to 25-35% in the dehydrated state, the beta-sheet content rises from 0 to 10-20%, and unordered structures increase from 40% to 50-60%. We have also investigated another model protein, hen egg-white lysozyme, and confirmed that it too undergoes a significant alteration of the secondary structure upon lyophilization. The extent of this structural reorganization has been found to be insensitive to the pH of the aqueous solution prior to lyophilization from pH 1.9 to 5.1, even though the thermal transition temperature (T-m) in aqueous solution over this range varies by 30 degrees C.