使用超高效液相色谱-质谱联用技术和免疫测定法对治疗领域抗体进行绝对定量。
Absolute quantification of a therapeutic domain antibody using ultra-performance liquid chromatography-mass spectrometry and immunoassay.
作者信息
Szapacs Matthew E, Urbanski James J, Kehler Jonathan R, Wilson Robert, Boram Sharon L, Hottenstein Charles S, Citerone David R
机构信息
Worldwide Bioanalysis, Drug Metabolism & Pharmacokinetics, GlaxoSmithKline, 709 Swedeland Rd, King of Prussia, PA 19406, USA.
出版信息
Bioanalysis. 2010 Sep;2(9):1597-608. doi: 10.4155/bio.10.70.
BACKGROUND
Domain antibodies (dAbs; ∼10-15 kDa) are made up of the variable heavy chain or the variable light chain of the antibody structure, and retain binding capability. dAbs have proved difficult to detect in plasma using immunoassay without specific antibodies raised against the dAb.
RESULTS
A sensitive and selective UPLC-MS/MS method for the absolute quantification of a dAb in monkey plasma was developed (range: 1 to 500 ng/ml) without the need for a specific capture antibody. This method was used to analyze pharmacokinetic studies early on in drug development. Furthermore, an immunoassay was developed and the pharmacokinetic samples were reanalyzed.
CONCLUSION
The two assays show good correlation (r(2) = 0.92), giving confidence in using either method for quantification of the dAb.
背景
结构域抗体（dAbs；约10 - 15 kDa）由抗体结构的可变重链或可变轻链组成，并保留结合能力。事实证明，在没有针对dAb产生的特异性抗体的情况下，使用免疫测定法很难在血浆中检测到dAbs。
结果
开发了一种灵敏且具有选择性的超高效液相色谱 - 串联质谱（UPLC-MS/MS）方法，用于在无需特异性捕获抗体的情况下对猴血浆中的dAb进行绝对定量（范围：1至500 ng/ml）。该方法在药物开发早期用于分析药代动力学研究。此外，还开发了一种免疫测定法，并对药代动力学样本进行了重新分析。
结论
这两种测定方法显示出良好的相关性（r(2) = 0.92），这使得对使用任何一种方法进行dAb定量都有信心。
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