National Taiwan Ocean University Institutional Repository:Item 987654321/27215
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题名: Measurement of Poly(ethylene glycol) by Cell-Based Anti-poly(ethylene glycol) ELISA
作者: Kuo-Hsiang Chuang;Shey-Cherng Tzou;Ta-Chun Cheng;Chien-Han Kao;Wei-Lung Tseng;Jentaie Shiea;Kuang-Wen Liao;Yun-Ming Wang;Ya-Chen Chang;Bo-Jyun Huang;Chang-Jer Wu;Pei-Yu Chu;Steve R. Roffler;Tian-Lu Cheng
贡献者: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
日期: 2010
上传时间: 2011-10-21T02:23:59Z
出版者: Anal Chem
摘要: Abstract:Poly(ethylene glycol) (PEG) is increasingly used in clinical and experimental medicine. However, quantification of PEG and PEGylated small molecules remains laborious and unsatisfactory. In this report, we stably expressed a functional anti-PEG antibody on the surface of BALB 3T3 cells (3T3/αPEG cells) to develop a competitive enzyme-linked immunosorbent assay (ELISA) for PEG quantification. The αPEG cell-coated plate bound biotinylated PEG5K (CH3-PEG5K-biotin) and CH3-PEG5K-131I more effectively than did a traditional anti-PEG antibody-coated plate. Competitive binding between PEG (2, 5, 10, or 20 kDa) and a known amount of CH3-PEG5K-biotin allowed construction of a reproducible competition curve. The αPEG cell-based competition ELISA measured small molecules derivatized by PEG2K, PEG5K, PEG10K, PEG20K, and PEG5K at concentrations as low as 58.6, 14.6, 3.7, 3.7, and 14.6 ng/mL, respectively. Notably, the presence of serum or bovine serum albumin enhanced PEG measurement by the αPEG cell-based competition ELISA. Finally, we show here that the αPEG cell-based competition ELISA accurately delineated the pharmacokinetics of PEG5K, comparable to those determined by direct measurement of radioactivity in blood after intravenous injection of CH3-PEG5K-131I into mice. This quantitative strategy may provide a simple and sensitive method for quantifying PEG and PEGylated small molecules in vivo.
關聯: 82(6), pp.2355–2362
URI: http://ntour.ntou.edu.tw/handle/987654321/27215
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