Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance
5 affiliations
- 1Department of Pharmacy, Huashan Hospital, Fudan University, Shanghai, 200030, China
- 2Department of Pharmacy, Jing'an District Central Hospital of Shanghai, Fudan University, Shanghai, 200040, China
- 3Department of Pharmacology, School of Basic Medical Sciences, Department of Pharmacy, Pudong Hospital, State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, 200032, China
- 4School of Pharmaceutical Sciences, Key Laboratory of Smart Drug Delivery, Ministry of Education & MOE Innovative Center for New Drug Development of Immune Inflammatory Diseases, Fudan University, Shanghai, 201203, China
- 5Department of Pharmacy, Huashan Hospital, Fudan University, Shanghai, 200030, China; Department of Pharmacy, Jing'an District Central Hospital of Shanghai, Fudan University, Shanghai, 200040, China; Corresponding author. Department of Pharmacy, Huashan Hospital, Fudan University, Shanghai, 200030, China.
Abstract
Polyethylene glycol (PEG) is a widely used pharmaceutical excipient to improve drug solubility and prolong in vivo circulation duration. Although it was initially considered non-immunogenic, recent studies have confirmed the ubiquitous presence of anti-PEG antibodies pre-existing in human, influencing the efficacy and safety of multiple PEGylated medicines. Due to the contradictions of current anti-PEG antibody detection methods and unclear induction mechanism of human pre-existing anti-PEG antibodies, the actual prevalence, binding features, and biological effects of pre-existing anti-PEG antibodies remain elusive, hampering the clinical implementation of personalized PEGylated therapeutics, as well as the rational design of evasion strategies and the development of innovative PEG materials and PEGylated nanocarriers. Herein, we established a robust enzyme-linked immunosorbent assay (ELISA)-based methodology for the detection of human pre-existing anti-PEG antibodies, and defined the positive threshold based on complement activation. Mechanistic analysis revealed that human pre-existing anti-PEG IgG predominantly recognized the PEG backbone, while anti-PEG IgM specifically bound to the methoxy end group. Notably, the weak binding affinity of human pre-existing anti-PEG IgG limited its ability to induce complement activation or promote capture of PEGylated liposomes by mononuclear phagocyte system, in contrast to IgM. Collectively, this study not only provided a reliable detection strategy for human pre-existing anti-PEG antibodies, but also revealed the distinct binding patterns, affinities, and biological effects of pre-existing anti-PEG IgM and IgG, with important implications for the safe and rational applications of PEGylated therapeutics.
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BibTeX
@article{fu2026standardizing,
title = {{Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance}},
author = {Jiaru Fu and Tianhao Ding and Guanghui Li and Ercan Wu and Anqi Wei and Mengyuan Liu and Bin Wang and Changyou Zhan},
journal = {Journal of Pharmaceutical Analysis},
year = {2026},
month = sep,
doi = {10.1016/j.jpha.2026.101609},
url = {https://doi.org/10.1016/j.jpha.2026.101609},
note = {CC BY},
}
RIS
TY - JOUR
TI - Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance
AU - Jiaru Fu
AU - Tianhao Ding
AU - Guanghui Li
AU - Ercan Wu
AU - Anqi Wei
AU - Mengyuan Liu
AU - Bin Wang
AU - Changyou Zhan
PY - 2026
DA - 2026/09/01
JO - Journal of Pharmaceutical Analysis
DO - 10.1016/j.jpha.2026.101609
UR - https://doi.org/10.1016/j.jpha.2026.101609
AB - Polyethylene glycol (PEG) is a widely used pharmaceutical excipient to improve drug solubility and prolong in vivo circulation duration. Although it was initially considered non-immunogenic, recent studies have confirmed the ubiquitous presence of anti-PEG antibodies pre-existing in human, influencing the efficacy and safety of multiple PEGylated medicines. Due to the contradictions of current anti-PEG antibody detection methods and unclear induction mechanism of human pre-existing anti-PEG antibodies, the actual prevalence, binding features, and biological effects of pre-existing anti-PEG antibodies remain elusive, hampering the clinical implementation of personalized PEGylated therapeutics, as well as the rational design of evasion strategies and the development of innovative PEG materials and PEGylated nanocarriers. Herein, we established a robust enzyme-linked immunosorbent assay (ELISA)-based methodology for the detection of human pre-existing anti-PEG antibodies, and defined the positive threshold based on complement activation. Mechanistic analysis revealed that human pre-existing anti-PEG IgG predominantly recognized the PEG backbone, while anti-PEG IgM specifically bound to the methoxy end group. Notably, the weak binding affinity of human pre-existing anti-PEG IgG limited its ability to induce complement activation or promote capture of PEGylated liposomes by mononuclear phagocyte system, in contrast to IgM. Collectively, this study not only provided a reliable detection strategy for human pre-existing anti-PEG antibodies, but also revealed the distinct binding patterns, affinities, and biological effects of pre-existing anti-PEG IgM and IgG, with important implications for the safe and rational applications of PEGylated therapeutics.
SN - 2095-1779
SN - 2214-0883
ER -