This assay primarily detects antibodies directed against the polyoxyethylene backbone of PEG.
Contents of Kit
No.
Components
Size
Storage Conditions
1
PEG-BSA Coated Plate
12×8 wells
-20°C
2
Anti-Monkey IgG HRP Stock (lyophilized)
1
-20°C
3
Reference Stock (lyophilized)
1
-20°C
4
20× HRP PEG Wash
50 ml
2-8°C
5
HRP PEG Diluent
50 ml
2-8°C
6
TMB
11 ml
2-8°C
7
Stop Solution
11 ml
2-8°C
Storage
The reference stock, HRP conjugate and the PEG-BSA coated plate should be stored at -20°C. All remaining kit components should be stored at 4°C. The microtiter plate should be kept in a sealed bag with desiccant. Kits will remain stable for six months from the date of purchase provided that the components are stored as described.
General Description
Attachment of polyethylene glycol (PEG) chains to therapeutic biologic agents, a process referred to as PEGylation, prolongs the circulating half-life of the modified protein by slowing proteolytic degradation and by masking it from the immune system. However, it has been reported that repeated injections of PEGylated proteins can induce anti-PEG antibodies that increase the rate of clearance and decrease drug efficacy (accelerated blood clearance, or ABC phenomenon). In our own studies, we find that a single injection of PEGylated protein induced high anti-PEG IgG titers (refer to the "Sample Preparation" section.) To aid research in this key area, we have developed a monkey anti-PEG IgG ELISA kit.
Standard Curve
A typical standard curve with optical density readings at 450nm on the Y-axis against log10 anti-PEG IgG concentrations on the X-axis is shown below. This curve is for the purpose of illustration only and should not be used to calculate unknowns.
Citations
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Background
PEG is a synthetic polymer widely used in various biomedical applications, such as drug delivery systems, medical implants, and diagnostics, due to its biocompatibility and versatility. PEGylation, the process of covalently attaching PEG molecules to therapeutic proteins and nanoparticles, has been widely employed to enhance their stability, solubility, and circulation time in the body. However, these biomedicines are often immunogenic, triggering the emergence of antidrug antibodies (ADAs). Repeated injections of PEGylated proteins have been reported to induce anti-PEG antibodies, including anti-PEG IgG and IgM, which have been shown to account for the loss of efficacy due to accelerated blood clearance of the drug (the ABC phenomenon) and hypersensitivity reactions (HSRs) leading to fatal allergic reactions. Understanding the presence and concentration of anti-PEG IgG antibodies in individuals is crucial for assessing the potential immunogenicity of PEGylated products. This information helps researchers and pharmaceutical companies evaluate the safety and efficacy profiles of these products, identify individuals at risk of adverse immune reactions, and guide treatment decisions. Detection and quantification of anti-PEG IgG antibodies are typically performed using immunoassay techniques, such as ELISA (enzyme-linked immunosorbent assay). We manufacture the anti-PEG IgG ELISA Kit for measurement of anti-PEG IgG in monkey serum or plasma. It allows efficient and convenient testing, enabling laboratories to process multiple samples simultaneously and obtain reliable results.
Alternative Names
Anti-PEG IgG ELISA Anti-Polyethylene Glycol IgG ELISA Anti-Polyethylene Glycol Immunoglobulin G ELISA Anti-PEG Immunoglobulin G ELISA Anti-PEG IgG ELISA Kit Anti-Polyethylene Glycol IgG ELISAKit Anti-Polyethylene Glycol Immunoglobulin G Kit Anti-PEG Immunoglobulin G ELISA Kit
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References
Anti-PEG immunity: emergence, characteristics, and unaddressed questions
Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology
The modification of protein and nanoparticle therapeutics with polyethylene glycol (PEG), a flexible, uncharged, and highly hydrophilic polymer, is a widely adopted approach to reduce RES clearance, extend circulation time, and improve drug efficacy. Nevertheless, an emerging body of literature, generated by numerous research groups, demonstrates that the immune system can produce antibodies that specifically bind PEG, which can lead to the 'accelerated blood clearance' of PEGylated therapeutics. In animals, anti-PEG immunity is typically robust but short-lived and consists of a predominantly anti-PEG IgM response. Rodent studies suggest that the induction of anti-PEG antibodies (α-PEG Abs) primarily occurs through a type 2 T-cell independent mechanism. Although anti-PEG immunity is less well-studied in humans, the presence of α-PEG Abs has been correlated with reduced efficacy of PEGylated therapeutics in clinical trials. The prevalence of anti-PEG IgG and reports of memory immune responses, as well as the existence of α-PEG Abs in healthy untreated individuals, suggests that the mechanism(s) and features of human anti-PEG immune responses may differ from those of animal models. Many questions, including the incidence rate of pre-existing α-PEG Abs and immunological mechanism(s) of α-PEG Ab formation in humans, must be answered in order to fully address the potential complications of anti-PEG immunity.
Anti-PEG antibodies: Current situation and countermeasures
Polyethylene glycol (PEG) is continuing to be a star molecular to increase hydrophilicity and to lengthen the blood circulation after conjugation with therapeutic proteins, nucleotides and nanocarriers. In contrary to the inherent idea, anti-PEG antibodies have been discovered in multiple species after treatment with PEGylated therapeutics. It is even worse that pre-existing anti-PEG antibodies demonstrate increasing prevalence in healthy people without prior exposure to PEGylated therapeutics but reasons underlying await to be clarified. Anti-PEG antibodies, no matter induced or pre-existing, would exert a series of biological effects including but not limited to accelerated blood clearance (ABC) phenomenon, and hypersensitivity reactions after encountering with PEGylated therapeutics, which potentially cause hazards to efficacy and safety of clinical medication. Considering the wide application of PEG in pharmaceutical field, mechanistic understanding on anti-PEG antibodies production and strategies to manage the consequent biological effects are desperately needed. In this review, the prevalence and possible mechanism of anti-PEG antibodies production were outlined. A series of factors that affecting our understanding the impacts of anti-PEG antibodies on in vivo biological effects against PEGylated therapeutics and potential countermeasures were highlighted.
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