Development of an Anti-Methoxy Poly(ethylene glycol) (伪-mPEG) Cell-Based Capture System to Measure mPEG and mPEGylated Molecules

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• 作者：Kuo-Hsiang Chuang ; Chien-Han Kao ; Steve R. Roffler ; Ssu-Jung Lu ; Ta-Chun Cheng ; Yun-Ming Wang ; Chih-Hung Chuang ; Yuan-Chin Hsieh ; Yeng-Tseng Wang ; Jaw-Yuan Wang ; Kuo-Yi Weng ; Tian-Lu Cheng
• 刊名：Macromolecules
• 出版年：2014
• 出版时间：October 14, 2014
• 年：2014
• 卷：47
• 期：19
• 页码：6880-6888
• 全文大小：426K
• ISSN：1520-5835

文摘

Quantitative pharmacokinetic analysis of methoxy-poly(ethylene glycol) (mPEG) and mPEGylated molecules is important for clinical drug development. Here we developed sensitive sandwich and competitive ELISAs by expressing an anti-mPEG antibody on the surface of fibroblasts for effective capture of mPEG molecules in biological samples. 伪-mPEG sandwich ELISA could quantify the higher-molecular-weight of mPEG (2, 5, and 20 kDa) and mPEGylated molecules. 伪-mPEG cell-based competitive ELISA was developed to measure the lower-molecular-weight of mPEG molecules (559, 750, and 1000 Da) at nanomolar levels. In addition, 伪-mPEG cell-based ELISA was unaffected by the presence of 10% human serum or murine serum. We further demonstrate that the 伪-mPEG cell-based ELISA determined similar pharmacokinetics of mPEG_5K as traditional gamma counting of ¹³¹I-mPEG_5K. The 伪-mPEG cell-based ELISA may provide an accurate, high sensitivity and easy-to-use tool for directly measuring mPEG and mPEGylated molecules in complex biological samples to accelerate the clinical development of mPEG drugs.