Multiplexed Evaluation of Serum and CSF Pharmacokinetics of Brain-Targeting Single-Domain Antibodies Using a NanoLC鈥揝RM-ILIS Method

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• 作者：Arsalan S. Haqqani ; Nadia Caram-Salas ; Wen Ding ; Eric Brunette ; Christie E. Delaney ; Ewa Baumann ; Eve Boileau ; Danica Stanimirovic
• 刊名：Molecular Pharmaceutics
• 出版年：2013
• 出版时间：May 6, 2013
• 年：2013
• 卷：10
• 期：5
• 页码：1542-1556
• 全文大小：717K
• 年卷期：v.10,no.5(May 6, 2013)
• ISSN：1543-8392

文摘

FC5 and FC44 are single-domain antibodies (V_HHs), selected by functional panning of phage-display llama V_HH library for their ability to internalize human brain endothelial cells (BEC) and to transmigrate the in vitro BBB model. Quantification of brain delivery of FC5 and FC44 in vivo was challenging using classical methods because of their short plasma half-life and their loss of functionality with radioactive labeling. A highly sensitive (detection limit <2 ng/mL) and specific SRM-ILIS method to detect and quantify unlabeled V_HHs in multiplexed assays was developed and applied to comparatively evaluate brain delivery of FC5 and FC44, and two control V_HHs, EG2 and A20.1. FC5 and FC44 compared to control V_HHs demonstrated significantly (p < 0.01) enhanced transport (50鈥?00-fold) across rat in vitro BBB model as well as in vivo brain targeting assessed by optical imaging. The multiplexed SRM-ILIS analyses of plasma and CSF levels of codosed V_HHs demonstrated that while all 4 V_HHs have similar blood pharmacokinetics, only FC5 and FC44 show elevated CSF levels, suggesting that they are potential novel carriers for delivery of drugs and macromolecules across the BBB.

Keywords:

blood鈭抌rain barrier; single-domain antibodies; CSF; pharmacokinetics; MRM; SRM; mass spectrometry; drug delivery