Sensitive quantification of PEGylated compounds by second-generation anti-poly(ethylene glycol) monoclonal antibodies

Yu Cheng Su, Bing Mae Chen, Kuo Hsiang Chuang, Tian Lu Cheng, Steve R. Roffler

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Poly(ethylene glycol) (PEG) is often attached to compounds to increase serum half-life, reduce immunogenicity, and enhance bioavailability. Accurate and sensitive quantification of PEG conjugates is critical for product development, pharmacokinetic measurements, and efficacy studies. However, PEGylated compounds can be difficult to quantify due to epitope masking by PEG. We previously generated two monoclonal antibodies to PEG (AGP3, IgM and E11, IgG) for quantitative detection of PEGylated proteins. We now report the identification of two second-generation mAbs to PEG (AGP4, IgM and 3.3, IgG) that bind to the repeating subunits of the PEG backbone and facilitate more sensitive quantification of a wider range of PEGylated compounds. A sandwich ELISA in which AGP4/3.3-biotin was employed as the capture/detection antibodies allowed quantification of PEG-Qdot 525 with 14-50-fold greater sensitivity than the original AGP3/E11 combination. Pegasys (PEG-interferon alpha-2a), PEG-Intron (PEG-interferon alpha-2b), Neulasta (PEG-G-CSF), and Lipo-Dox (PEGylated liposomal doxorubicin) could also be quantified with low ng/mL detection limits. The assay tolerated the presence of 50% human serum or 20% free PEG molecules. These new anti-PEG antibodies appear useful for qualitative and quantitative analysis of a wide range of PEGylated compounds.

Original languageEnglish
Pages (from-to)1264-1270
Number of pages7
JournalBioconjugate Chemistry
Volume21
Issue number7
DOIs
Publication statusPublished - Jul 21 2010
Externally publishedYes

Fingerprint

Monoclonal antibodies
interferon alfa-2b
Ethylene Glycol
Polyethylene glycols
Immunoglobulin M
Immunoglobulin G
Monoclonal Antibodies
Granulocyte Colony-Stimulating Factor
Biotin
Serum
Biological Availability
Half-Life
Limit of Detection
Epitopes
Anti-Idiotypic Antibodies
Pharmacokinetics
Enzyme-Linked Immunosorbent Assay
Antibodies
Interferons
Proteins

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Sensitive quantification of PEGylated compounds by second-generation anti-poly(ethylene glycol) monoclonal antibodies. / Su, Yu Cheng; Chen, Bing Mae; Chuang, Kuo Hsiang; Cheng, Tian Lu; Roffler, Steve R.

In: Bioconjugate Chemistry, Vol. 21, No. 7, 21.07.2010, p. 1264-1270.

Research output: Contribution to journalArticle

Su, Yu Cheng ; Chen, Bing Mae ; Chuang, Kuo Hsiang ; Cheng, Tian Lu ; Roffler, Steve R. / Sensitive quantification of PEGylated compounds by second-generation anti-poly(ethylene glycol) monoclonal antibodies. In: Bioconjugate Chemistry. 2010 ; Vol. 21, No. 7. pp. 1264-1270.
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