Quantification of active infliximab in human serum with liquid chromatography-tandem mass spectrometry using a tumor necrosis factor alpha -based pre-analytical sample purification and a stable isotopic labeled infliximab bio-similar as internal standard: A target-based, sensitive and cost-effective method
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- 作者：Mohsin El Amrani ; m.elamrani@umcutrecht.nl" class="auth_mail" title="E-mail the corresponding author ; mohsin.elamrani@gmail.com" class="auth_mail" title="E-mail the corresponding author ; Marcel P.H. van den Broek ; Camiel Gö ; bel ; Erik M. van Maarseveen
- 关键词：Infliximab ; Immuno-affinity ; Streptavidin ; Biotin ; TNF-α
- 刊名：Journal of Chromatography A
- 出版年：2016
- 出版时间：8 July 2016
- 年：2016
- 卷：1454
- 期：Complete
- 页码：42-48
- 全文大小：1283 K
文摘
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A selective purification and quantification of Infliximab in human serum is proposed.
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The purification is based on immuno-affinity using biotinylated TNF-alpha captured onto a streptavidin coated 96 well plate.
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Quantification is performed using LC–MS/MS on one signature peptide after tryptic digestion.
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The method was validated according to EMA guidelines, has a lower limit of quantification of 0.5 μg/mL using 2 μL serum.
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Cross validation against reference ELISA method showed a strong agreement of r2 = 0.95.