| Measurement of poly(ethylene glycol) by cell-based anti-poly(ethylene glycol) ELISA. | |
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MedLine Citation:
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PMID: 20178318 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Analytical chemistry Volume: 82 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2010 Mar |
Date Detail:
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Created Date: 2010-03-12 Completed Date: 2010-06-08 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 2355-62 Citation Subset: IM |
Affiliation:
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Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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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