標題: Measurement of Poly(ethylene glycol) by Cell-Based Anti-poly(ethylene glycol) ELISA
作者: Chuang, Kuo-Hsiang
Tzou, Shey-Cherng
Cheng, Ta-Chun
Kao, Chien-Han
Tseng, Wei-Lung
Shiea, Jentaie
Liao, Kuang-Wen
Wang, Yun-Ming
Chang, Ya-Chen
Huang, Bo-Jyun
Wu, Chang-Jer
Chu, Pei-Yu
Roffler, Steve R.
Cheng, Tian-Lu
生物科技學系
Department of Biological Science and Technology
公開日期: 15-三月-2010
摘要: 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/alpha PEG cells) to develop a competitive enzyme-linked immunosorbent assay (ELISA) for PEG quantification. The alpha-PEG cell-coated plate bound biotinylated PEG(5K) (CH(3)-PEG(5K)-biotin) and CH(3)-PEG(5K)-(131)I 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 CH(3)-PEG(5K)-biotin allowed construction of a reproducible competition curve. The alpha PEG cell-based competition ELISA measured small molecules derivatized by PEG(2K), PEG(5K), PEG(10K), PEG(20K), and PEG(5K) 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 alpha PEG cell-based competition ELISA. Finally, we show here that the alpha PEG cell-based competition ELISA accurately delineated the pharmacokinetics of PEG(5K), comparable to those determined by direct measurement of radioactivity in blood after intravenous injection of CH(3)-PEG(5K)-(131)I into mice. This quantitative strategy may provide a simple and sensitive method for quantifying PEG and PEGylated small molecules in vivo.
URI: http://dx.doi.org/10.1021/ac902548m
http://hdl.handle.net/11536/5707
ISSN: 0003-2700
DOI: 10.1021/ac902548m
期刊: ANALYTICAL CHEMISTRY
Volume: 82
Issue: 6
起始頁: 2355
結束頁: 2362
顯示於類別:期刊論文


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