AffinityImmuno · ELISA Kit
Anti PEG antibody ELISA (human IgM specific)
Cat.No
EL-141-PEG-hIGM
1X96 wells
- Antigen
- Anti-PEG human IgM antibodies
- Reactivity
- human
- Detection range
- 62.5 ng/mL -1000 ng/mL
- Detection minimum
- 62.5 ng/mL
- Sample type
- Serum Plasma
- Assay time
- 2.5 hours
Product Overview
This immunogenicity assay measures the presence of human IgM in serum or plasma that bind to immobilized PEG using the indirect ELISA technique.
Introduction
Polyethylene glycol (PEG) chains are often used to modify therapeutic biologic agents in order to prolong the circulating half-life of the modified protein by slowing proteolytic degradation. It has been reported that repeat injections of PEGylated proteins can induce anti-PEG antibodies. Anti-PEG antibodies can result in rapid clearance and decreased drug efficacy (accelerated blood clearance, or ABC, phenomenon).
Principle of the Assay
This immunogenicity assay measures the presence of human IgM in serum or plasma that bind to immobilized PEG using the indirect ELISA technique. The supplied 96 well microplate is pre-coated with PEG. After washing, quality control and test samples are pipetted into the appropriate wells. Anti-PEG antibodies bind the immobilized PEG. After washing, detection antibody (anti human IgM Peroxidase) is added. Any unbound antibody enzyme reagent is removed with a final wash and a substrate solution is added to the wells for color development. Color development is proportional to the amount of anti-PEG IgM.
Related assays
Also available in other species/isotype specificities: human IgG, mouse IgG, and mouse IgM.
Related reading
For context on anti-PEG IgM prevalence and clinical significance in human samples, see our 2026 PEG immunogenicity review.
Target background
Polyethylene glycol (PEG) chains are often used to modify therapeutic biologic agents in order to prolong the circulating half-life of the modified protein. It has been reported that repeat injections of PEGylated proteins can induce anti-PEG antibodies.
Specifications
- Assay precision
- Intra-assay CV <10%. Inter-assay CV <10%
- Storage
- Stable at -20°C for 1 year
- Size
- 1X96 wells
- Antigen
- Anti-PEG human IgM antibodies
- Reactivity
- human
- Detection range
- 62.5 ng/mL -1000 ng/mL
- Detection minimum
- 62.5 ng/mL
- Method
- Direct sandwich ELISA
- Detection method
- Peroxidase / OD450
- Application
- Quantification of human IgM antibodies to PEG
- Plate format
- Strip
- Sample type
- Serum Plasma
- Sample volume
- 15ul
- Assay time
- 2.5 hours
- Specificity
- Anti-PEG antibodies
- Preservative
- none
- Safety warning
- Research Use Only. Follow instructions.
Kit Components
- Coated microtiter plate, 96 wells
- QC samples - 6x250ul
- 1X assay buffer - 50ml
- 10X wash buffer concentrate - 50ml
- 1000X detection reagent - 17ul
- TMB - 12ml
- TMB stop solution - 12ml
- Plate sealers - 3
Assay Information
Reagent preparation
Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation Dilute wash buffer concentrate with deionized water 1/10 before use (for example add 20mL concentrate to 180mL deionized water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11μl concentrate to 11ml of assay buffer). Mix well. The following is an example calibrator curve.
Results calculation
1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
Sample collection
This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20°C for up to 1 year.
Sample preparation
Dilute test samples 1/5 with assay buffer before use (for example add 50μl of test sample to 200μl assay buffer). Mix well.