Document Detail


Measurement of poly(ethylene glycol) by cell-based anti-poly(ethylene glycol) ELISA.
MedLine Citation:
PMID:  20178318     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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/alphaPEG cells) to develop a competitive enzyme-linked immunosorbent assay (ELISA) for PEG quantification. The alphaPEG 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 alphaPEG 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 alphaPEG cell-based competition ELISA. Finally, we show here that the alphaPEG 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.
Authors:
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
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  82     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2010 Mar 
Date Detail:
Created Date:  2010-03-12     Completed Date:  2010-06-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2355-62     Citation Subset:  IM    
Affiliation:
Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies / immunology*
BALB 3T3 Cells
Cattle
Enzyme-Linked Immunosorbent Assay / methods*
Female
Mice
Polyethylene Glycols / analysis*,  pharmacokinetics*
Serum / chemistry
Serum Albumin, Bovine / chemistry
Chemical
Reg. No./Substance:
0/Antibodies; 0/Polyethylene Glycols; 0/Serum Albumin, Bovine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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