Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length

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

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1408-1413
Number of pages6
JournalBioconjugate Chemistry
Volume24
Issue number8
DOIs
Publication statusPublished - Aug 21 2013

Fingerprint

Interferons
Antibodies
Polyethylene glycols
Enzyme-Linked Immunosorbent Assay
Epitopes
Proteins
Pharmacokinetics
Ethylene Glycol
Masks
Serum
Introns
Half-Life
Limit of Detection
Assays

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology
  • Medicine(all)

Cite this

Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length. / 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.

In: Bioconjugate Chemistry, Vol. 24, No. 8, 21.08.2013, p. 1408-1413.

Research output: Contribution to journalArticle

Cheng, TC, Chuang, KH, Chen, M, Wang, HE, Tzou, SC, Su, YC, Chuang, CH, Kao, CH, Chen, BM, Chang, LS, Roffler, SR & Cheng, TL 2013, 'Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length', Bioconjugate Chemistry, vol. 24, no. 8, pp. 1408-1413. https://doi.org/10.1021/bc3006144
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. / Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length. In: Bioconjugate Chemistry. 2013 ; Vol. 24, No. 8. pp. 1408-1413.
@article{8586156dfdf44457b375dc87e1f4a775,
title = "Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length",
abstract = "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.",
author = "Cheng, {Ta Chun} and Chuang, {Kuo Hsiang} and Michael Chen and Wang, {Hsin Ell} and Tzou, {Shey Cherng} and Su, {Yu Cheng} and Chuang, {Chih Hung} and Kao, {Chien Han} and Chen, {Bing Mae} and Chang, {Long Sen} and Roffler, {Steve R.} and Cheng, {Tian Lu}",
year = "2013",
month = "8",
day = "21",
doi = "10.1021/bc3006144",
language = "English",
volume = "24",
pages = "1408--1413",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "8",

}

TY - JOUR

T1 - Sensitivity of PEGylated interferon detection by anti-polyethylene glycol (PEG) antibodies depends on PEG length

AU - Cheng, Ta Chun

AU - Chuang, Kuo Hsiang

AU - Chen, Michael

AU - Wang, Hsin Ell

AU - Tzou, Shey Cherng

AU - Su, Yu Cheng

AU - Chuang, Chih Hung

AU - Kao, Chien Han

AU - Chen, Bing Mae

AU - Chang, Long Sen

AU - Roffler, Steve R.

AU - Cheng, Tian Lu

PY - 2013/8/21

Y1 - 2013/8/21

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84883154050&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84883154050&partnerID=8YFLogxK

U2 - 10.1021/bc3006144

DO - 10.1021/bc3006144

M3 - Article

C2 - 23837865

AN - SCOPUS:84883154050

VL - 24

SP - 1408

EP - 1413

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 8

ER -