Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms.
Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms.
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关联明确的寡糖结构对 IgG1-Fc 糖型物理稳定性的影响。
DOI：
10.1016/j.xphs.2015.10.014
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发表时间：
2016
影响因子：
3.8
通讯作者：
Volkin,DavidB
中科院分区：
文献类型：
--
作者：
More,ApurvaS;Toprani,VishalM;Okbazghi,SolomonZ;Kim,JaeH;Joshi,SangeetaB;Middaugh,CRussell;Tolbert,ThomasJ;Volkin,DavidB
作为本期特刊系列文章的一部分，本文描述了4种定义良好的IgG1-Fc糖型作为生物相似性分析的模型系统（高甘露糖- fc， Man5-Fc， GlcNAc-Fc和N297Q-Fc糖基化），本工作的重点是比较它们的物理性质。聚乙二醇沉淀法（pH值为4.5、6.0）发现n297连接的低聚糖结构尺寸减小，表观溶解度（热力学活性）降低，差示扫描量热法（pH值为4.5）发现构象稳定性降低。使用多种高通量生物物理技术，在2种配方（NaCl和蔗糖）下，在广泛的温度范围（10°C-90°C）和pH（4.0-7.5）条件下，测量了Fc糖蛋白的物理稳定性。数据集用于构建3指数经验相图和雷达图，以可视化蛋白质结构稳定性区域。每种糖形式在蔗糖（相对于盐）配方中表现出更好的稳定性。HM-Fc和Man5-Fc的相对稳定性最高，其次是GlcNAc-Fc， N297Q-Fc的相对稳定性最低。因此，4种IgG1-Fc糖型的整体物理稳定性也显示出与低聚糖结构的相关性。这些数据集用于开发生物相似性分析的数学模型（如Kim等人在本期的配套文章中所述）。
As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. Each glycoform showed improved stability in the sucrose (vs. salt) formulation. The HM-Fc and Man5-Fc displayed the highest relative stability, followed by GlcNAc-Fc, with N297Q-Fc being the least stable. Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue).