標題: Sensitivity of PEGylated Interferon Detection by Anti-Polyethylene Glycol (PEG) Antibodies Depends on PEG Length
作者: Cheng, Ta-Chun
Chuang, Kuo-Hsiang
Chen, Michael
Wang, Hsin-Ell
Tzou, Shey-Cherng
Su, Yu-Cheng
Chuang, Chih-Hung
Kao, Chien-Han
Chen, Bing-Mae
Chang, Long-Sen
Roffler, Steve R.
Cheng, Tian-Lu
生物科技學系
Department of Biological Science and Technology
公開日期: 1-Aug-2013
摘要: Attachment of poly(ethylene glycol) to proteins can mask immune epitopes to increase serum half-life, reduce immunogenicity, and enhance in vivo biological efficacy. However, PEGylation mediated epitope-masking may also limit sensitivity and accuracy of traditional ELISA. We previously described an anti-PEG-based sandwich ELISA for universal assay of PEGylated molecules. Here, we compared the quantitative assessment of PEGylated interferons by anti-PEG and traditional anti-interferon sandwich ELISA. The detection limits for PEG-Intron (12k-PEG) and Pegasys (40k-PEG) were 1.9 and 0.03 ng/mL for anti-PEG ELISA compared to 0.18 and 0.42 ng/mL for traditional anti-interferon sandwich ELISA. These results indicate that the anti-PEG sandwich ELISA was insensitive to PEGylation mediated epitope-masking and the sensitivity increased in proportion to the length of PEG. By contrast, PEG-masking interfered with detection by traditional anti-interferon sandwich ELISA. Human and mouse serum did not affect the sensitivity of anti-PEG ELISA but impeded traditional anti-interferon sandwich ELISA. The anti-PEG sandwich ELISA was comparable to anti-interferon sandwich ELISA and radioassay of I-131-Pegasys in pharmacokinetic studies in mice. The anti-PEG sandwich ELISA provides a sensitive, accurate, and convenient quantitative measurement of PEGylated protein drugs.
URI: http://dx.doi.org/10.1021/bc3006144
http://hdl.handle.net/11536/22586
ISSN: 1043-1802
DOI: 10.1021/bc3006144
期刊: BIOCONJUGATE CHEMISTRY
Volume: 24
Issue: 8
起始頁: 1408
結束頁: 1413
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