Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance

Jiaru Fu; Tianhao Ding; Guanghui Li; Ercan Wu; Anqi Wei; Mengyuan Liu; Bin Wang; Changyou Zhan

doi:10.1016/j.jpha.2026.101609

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Jiaru Fu, Tianhao Ding, Guanghui Li, Ercan Wu, Anqi Wei, Mengyuan Liu, Bin Wang, Changyou Zhan. Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101609

Citation:

Jiaru Fu, Tianhao Ding, Guanghui Li, Ercan Wu, Anqi Wei, Mengyuan Liu, Bin Wang, Changyou Zhan. Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101609

Citation:

Jiaru Fu, Tianhao Ding, Guanghui Li, Ercan Wu, Anqi Wei, Mengyuan Liu, Bin Wang, Changyou Zhan. Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101609

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Standardizing pre-existing anti-PEG antibody detection: A key to clarifying their binding features and clinical relevance

doi: 10.1016/j.jpha.2026.101609

Jiaru Fu^a,
Tianhao Ding^{b,c
,
,},
Guanghui Li^c,
Ercan Wu^b,
Anqi Wei^b,
Mengyuan Liu^d,
Bin Wang^{a,c
,
,},
Changyou Zhan^{a,b,d
,
,}

a. Department of Pharmacy, Huashan Hospital, Fudan University, Shanghai, 200030, China;
b. Department 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;
c. Department of Pharmacy, Jing'an District Central Hospital of Shanghai, Fudan University, Shanghai, 200040, China;
d. School 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

Funds:

This work was supported by National Key Research and Development Program, China (Grant No.: 2024YFE0216000), the National Natural Science Fund of China (Grant Nos.: 82125035, 32330058, 82404525, and 82361168639), Shanghai Municipal Education Commission Major Project, China (Grant No.: 2021-01-07-00-07-E00081), and China Postdoctoral Science Foundation (Grant No.: GZC20240305).

Received Date: Jul. 26, 2025
Accepted Date: Mar. 16, 2026
Rev Recd Date: Mar. 16, 2026
Available Online: Mar. 18, 2026

Abstract

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.
- Pre-existing anti-PEG antibodies,
- IgM,
- IgG,
- Binding affinity,
- Biological effects

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References(50)

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