Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length.
|Title||Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length.|
|Publication Type||Journal Article|
|Year of Publication||2013|
|Authors||Cheng, TC, Chuang KH, Chen M, Wang HE, Tzou SC, Su YC, Chuang CH, Kao CH, Chen BM, Chang LS, Roffler SR, Cheng TL|
|Date Published||2013 Jul 9|
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 sensitivity of anti-PEG sandwich ELISA was insensitive to PEGylation mediated epitope-masking and 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.
|Short Title||Bioconjug Chem|