Differential scanning fluorimetry: Rapid screening of formulations that promote the stability of reference preparations
Differential scanning fluorimetry: Rapid screening of formulations that promote the stability of reference preparations
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DOI：
10.1016/j.jpba.2013.01.006
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发表时间：
2013-04-15
影响因子：
3.4
通讯作者：
Burns, Chris J.
中科院分区：
文献类型：
--
作者：
Malik, Kiran;Matejtschuk, Paul;Burns, Chris J.
在配制生物药用蛋白质时，其液态稳定性至关重要。此外，在制备生物标准物质时，需要测定冻干和重建后的稳定性。为了使辅料在水环境中的稳定性达到最佳值(如特梅尔特或变性点所示)，应评估不同辅料选择的影响。微量差示扫描量热法是一种成熟的用于这些应用的方法，但即使使用自动进样器也可能非常耗时。差示扫描荧光法(DSF)是一种测量染料结合到变性蛋白质疏水区域时的荧光的新技术。我们使用α-1-蛋白酶抑制剂(A1PI)作为模型系统来研究这些技术的适用性，发现两种方法在不同辅料的影响方面有相似的趋势。DSF是一种很有前途的方法，在所需生物材料的速度和数量方面具有优势，并且可以使用PCR仪器进行。(C)2013爱思唯尔B.V.保留所有权利。
When formulating a biopharmaceutical protein, its stability in the liquid state is critical. In addition, when preparing biological reference materials the stability, both when lyophilised and after reconstitution, needs to be determined. In order to optimise the stability in aqueous conditions (as indicated by Tmelt or denaturation point) the impact of different excipient choices should be evaluated. Micro differential scanning calorimetry is a well established method for these applications but can be time consuming even when an autosampler is used. Differential scanning fluorimetry (DSF) is a novel technique which measures the fluorescence of a dye when bound to the hydrophobic regions of a denatured protein. We have investigated these techniques for their suitability using alpha-1-protease inhibitor (A1PI) as a model system and found similar trends in terms of the impact of different excipients by both methods. DSF is a promising method and has advantages in terms of speed and quantities of biological material required and can be performed using a PCR instrument. (c) 2013 Elsevier B.V. All rights reserved.