Fluorescent molecular rotors as dyes to characterize polysorbate-containing IgG formulations.
Fluorescent molecular rotors as dyes to characterize polysorbate-containing IgG formulations.
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DOI：
10.1007/s11095-009-0020-2
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发表时间：
2010-02
影响因子：
3.7
通讯作者：
Jiskoot, Wim
中科院分区：
文献类型：
--
作者：
Hawe, Andrea;Filipe, Vasco;Jiskoot, Wim
关键词：
目的是评估主要对粘度敏感的荧光分子转子（DCVJ和CCVJ），用于表征含聚山梨醇酯的IgG制剂，并将其与极性敏感染料ANS、Bis-ANS和尼罗红进行比较。将含有聚山梨酯20或80的IgG制剂在低于聚集温度下强制降解，并通过稳态和时间分辨荧光以及HP-SEC与UV和荧光染料检测（Bis-ANS和CCVJ）进行分析。此外，相应地聚集治疗性蛋白质的商业蛋白质制剂（Enbrel®50 mg、Humira®40 mg和MabThera®100 mg），并用CCVJ荧光和HP-SEC分析。与（双）ANS和尼罗红类似，分子转子DCVJ和CCVJ在含聚山梨醇酯的缓冲液中显示出低背景荧光。时间分辨荧光实验证实了稳态荧光数据。DCVJ和CCVJ均显示出聚集IgG制剂的增强的荧光强度，并且适合于在稳态荧光和具有染料检测的HP-SEC（CCVJ）中表征含聚山梨醇酯的IgG制剂。CCVJ能够检测含聚山梨醇酯的市售产品Enbrel®50 mg、Humira®40 mg和MabThera®100 mg中的热诱导聚集。荧光分子转子是检测含聚山梨醇酯的IgG制剂中聚集的合适探针。本文的在线版本（doi：10.1007/s11095-009-0020-2）包含补充材料，可供授权用户使用。
The aim was to evaluate fluorescent molecular rotors (DCVJ and CCVJ), which are mainly sensitive to viscosity, for the characterization of polysorbate-containing IgG formulations and compare them to the polarity-sensitive dyes ANS, Bis-ANS and Nile Red. IgG formulations with polysorbate 20 or 80 were stressed below the aggregation temperature and analyzed by steady-state and time-resolved fluorescence and by HP-SEC with UV and fluorescent dye detection (Bis-ANS and CCVJ). Furthermore, commercial protein preparations of therapeutic proteins (Enbrel®50 mg, Humira®40 mg and MabThera®100 mg) were aggregated accordingly and analyzed with CCVJ fluorescence and HP-SEC. Contrarily to (Bis-)ANS and Nile Red, the molecular rotors DCVJ and CCVJ showed low background fluorescence in polysorbate-containing buffers. Time-resolved fluorescence experiments confirmed the steady-state fluorescence data. Both DCVJ and CCVJ showed enhanced fluorescence intensity for aggregated IgG formulations and were suitable for the characterization of polysorbate-containing IgG formulations in steady-state fluorescence and HP-SEC with dye detection (CCVJ). CCVJ was capable of detecting thermally induced aggregation in the commercial polysorbate-containing products Enbrel®50 mg, Humira®40 mg and MabThera®100 mg. Fluorescent molecular rotors are suitable probes to detect aggregation in polysorbate-containing IgG formulations. The online version of this article (doi:10.1007/s11095-009-0020-2) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Hawe, Andrea;Sutter, Marc;Jiskoot, Wim
通讯作者：
Jiskoot, Wim
影响因子：
2.9
作者：
KUNG, CE;REED, JK
通讯作者：
REED, JK
影响因子：
5
作者：
Castro, GR;Larson, BK;Kaplan, DL
通讯作者：
Kaplan, DL
影响因子：
3.8
作者：
Jones, LS;Randolph, TW;Carpenter, JF
通讯作者：
Carpenter, JF
DOI：
10.1016/j.bbrc.2005.01.164
发表时间：
2005-04-15
影响因子：
3.1
作者：
Fu, XM;Zhang, XF;Chang, ZY
通讯作者：
Chang, ZY