Both IgM and IgG Antibodies against Polyethylene Glycol Can Alter the Biological Activity of Methoxy Polyethylene Glycol-Epoetin Beta in Mice

Abstract

Pre-existing antibodies that bind polyethylene glycol are present in about 40% of healthy individuals. It is currently unknown if pre-existing anti-polyethylene glycol (PEG) antibodies can alter the bioactivity of pegylated drugs with a single long PEG chain, which represents the majority of newly developed pegylated medicines. Methoxy polyethylene glycol-epoetin beta (PEG-EPO) contains a single 30 kDa PEG chain and is used to treat patients suffering from anemia. We find that the pre-existing human anti-PEG IgM and IgG antibodies from normal donors can bind to PEG-EPO. The prevalence and concentrations of anti-PEG IgM and IgG antibodies were also higher in patients that responded poorly to PEG-EPO. Monoclonal anti-PEG IgM and IgG antibodies at concentrations found in normal donors blocked the biological activity of PEG-EPO to stimulate the production of new erythrocytes in mice and accelerated the clearance of 125 I-PEG-EPO, resulting in PEG-EPO accumulation primarily in the liver and spleen. Accelerated clearance by the anti-PEG IgG antibody was mediated by the Fc portion of the antibody. Importantly, infusing higher doses of PEG-EPO could compensate for the inhibitory effects of anti-PEG antibodies, suggesting that pre-existing anti-PEG antibodies can be "dosed through." Our study indicates that the bioactivity and therapeutic activity of PEG-EPO may be reduced in patients with elevated levels of pre-existing anti-PEG antibodies. New pegylated medicines with a single long PEG chain may also be affected in patients with high levels of anti-PEG antibodies.

Polyethylene Glycol Immunogenicity: Theoretical, Clinical, and Practical Aspects of Anti-Polyethylene Glycol Antibodies

“…This analysis suggests that pre-existing anti-PEG IgM is not important for ABC and that pegylated interferons and epoetin-beta are most susceptible to ABC induced by anti-PEG IgG antibodies. This is consistent with reports that pre-existing anti-PEG antibodies decrease the anti-viral activity of PEG-interferon-α by accelerating clearance from the blood via uptake into Kupffer cells in the liver and an increased incidence of anti-PEG antibodies in patients who do not respond to PEG-epoetin-beta for the treatment of anemia associated with chronic kidney disease. , …”

“…The amount of anti-PEG antibody required to induce ABC also depends on antibody affinity (Figure B). High-affinity antibodies induce ABC when the molar ratio of anti-PEG antibody exceeds pegylated compound by about 3–10. ,, However, a higher molar ratio is required for low-affinity antibodies: up to 15 low-affinity IgM antibodies and over 90 low-affinity IgG antibodies are required to induce strong ABC of PEG-epoetin-beta in mice . These counteracting factors make estimation of the effects of pre-existing anti-PEG antibodies especially difficult since they possess relatively low affinity for PEG.…”

“…Several excellent reviews cover PEG chemistry, , assay of pegylated compounds, and immunogenicity of pegylated medicines. ,− In the present review, we aim to clear up some misconceptions about PEG immunogenicity, realistically assess the impact of anti-PEG antibodies on pegylated medicines, and describe the specificity and binding behavior of anti-PEG antibodies. We survey the literature as well as draw from personal experience over the past 20 years on creating anti-PEG monoclonal antibodies, developing anti-PEG antibody assays, − assaying and discovering genetic markers for anti-PEG antibodies, , investigating the effects of anti-PEG antibodies on pegylated medicines, ,− and creating recombinant anti-PEG receptors and targeting molecules − to provide a framework to understand what makes a pegylated drugs immunogenic, how anti-PEG antibodies affect the efficacy and safety of pegylated medicines, and some experience with how anti-PEG antibodies behave in practice.…”

PEG: Will It Come Back to You? Polyethelyne Glycol Immunogenicity, COVID Vaccines, and the Case for New PEG Derivatives and Alternatives

“…Indeed, prior studies show that strong ABC is observed when the number of antibodies in circulation exceeds the number of PEGylated compounds ( Xu et al, 2022 ). This trend holds across most PEGylated compounds including proteins, liposomes, micelles, and polymeric nanoparticles and agrees with previous studies showing that three anti-PEG antibodies per PEGylated protein or about 10 anti-PEG antibodies per pegylated liposome are required for ABC ( Shiraishi et al, 2016 ; McSweeney et al, 2018 ; Chang et al, 2019 ). These findings suggest that only compounds dosed at very low molar concentrations ( e.g.…”

Flow cytometry analysis of anti-polyethylene glycol antibodies in human plasma

“…PEGylation is the process of either covalently or non-covalently linking a high-molecular-weight (MW) PEG to an agent (drug or therapeutic peptide/protein), and is commonly used to increase the serum half-life of drugs or proteins/peptides, improve efficacy, and reduce the immunogenicity of the proteins/peptides [ 3 ]. To date, at least 20 PEGylated therapeutics have been approved for use in humans [ 4 ] and most of them have been used in the treatment of various diseases for over a decade.…”