Sensitivity of PEGylated Interferon Detection by Anti-Polyethylene Glycol (PEG) Antibodies Depends on PEG Length

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• 作者：Ta-Chun Cheng ; Kuo-Hsiang Chuang ; Michael Chen ; Hsin-Ell Wang ; Shey-Cherng Tzou ; Yu-Cheng Su ; Chih-Hung Chuang ; Chien-Han Kao ; Bing-Mae Chen ; Long-Sen Chang ; Steve R. Roffler ; Tian-Lu Cheng
• 刊名：Bioconjugate Chemistry
• 出版年：2013
• 出版时间：August 21, 2013
• 年：2013
• 卷：24
• 期：8
• 页码：1408-1413
• 全文大小：254K
• 年卷期：v.24,no.8(August 21, 2013)
• ISSN：1520-4812

文摘

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 ¹³¹I-Pegasys in pharmacokinetic studies in mice. The anti-PEG sandwich ELISA provides a sensitive, accurate, and convenient quantitative measurement of PEGylated protein drugs.