Simple and rapid analysis of tocilizumab using HPLC-fluorescence detection method
Simple and rapid analysis of tocilizumab using HPLC-fluorescence detection method
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DOI：
10.1002/bio.3615
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发表时间：
2019-05-01
期刊：
影响因子：
2.9
通讯作者：
Nakashima, Kenichiro
中科院分区：
文献类型：
--
作者：
Takada, Makoto;Ohba, Yoshihito;Nakashima, Kenichiro
我们开发了一种新的方法，使用高效液相色谱法（HPLC）与荧光检测测定托珠单抗（TCZ），经过一个简单和快速的前处理。TCZ与IgG1同属免疫球蛋白G亚类1（Immunocytulin G subclass 1），利用该检测方法可以在不发生抗原抗体反应的情况下将TCZ与IgG1分离。这些抗体的分离通过用含有碱（如三甲基胺和三乙胺）的有机溶剂预处理来实现。这些碱基对TCZ分离的影响与碱基的疏水性有关，而与静电荷无关。结果表明，尽管抗体的氨基酸序列差异很小，但由于结构变化，抗体的表面电荷发生了变化。该方法可用于TCZ抗体亚类的分离，并可用于血中TCZ的检测。
We developed a novel assay using high-performance liquid chromatography (HPLC) with fluorescence detection for the determination of tocilizumab (TCZ), after it has undergone a facile and rapid pretreatment. TCZ belongs to the same subclass as IgG1 (Immunoglobulin G subclass 1), and we could separate TCZ from IgG1 without antigen-antibody reactions, with the novel detection method. The separation of these antibodies was achieved by pretreatment with an organic solvent containing a base, such as trimethylamine and triethylamine. The effect of these bases on the separation of TCZ is related to the hydrophobicity of the base rather than the electrostatic charge. The results indicated that the surface charge of antibodies changed because of the structural change, even though the difference in the amino acid sequences of the antibodies was very low. Our method is available for the separation of the antibody subclasses, and it would be useful to assay TCZ in blood.