Cat. No.
MAK-0014
Product Name
Monkey Anti-PEG IgG ELISA kit (ADA)
Storage
2–8°C.
Principle
Indirect ELISA
Intended Use
This kit is for research use only. Under no circumstances should it be used for therapeutic or human diagnostic applications.
Principle Of The Assay
The assay uses immobilized mono-mPEGylated BSA (20 kDa PEG chain) as the capture antigen (coated on microtiter wells) and horseradish peroxidase (HRP) conjugated anti-monkey IgG monoclonal antibody (LDI clone 6D11-21) for detection. Serum or plasma samples are diluted and incubated alongside standards in the microtiter wells for one hour. The wells are subsequently washed. HRP conjugate is added and incubated for 45 minutes. Anti-PEG IgG molecules are thus sandwiched between immobilized PEG and the detection antibody conjugate. The wells are then washed to remove unbound HRP-conjugate. TMB reagent is added and incubated for 20 minutes at room temperature. This results in the development of a blue color. Color development is stopped by the addition of Stop Solution; changing the color to yellow, and optical density is measured spectrophotometrically at 450 nm. The concentration of anti-PEG IgG is proportional to the absorbance at 450 nm and is derived from a standard curve. This assay primarily detects antibodies directed against the polyoxyethylene backbone of PEG. Studies at Mabioway Co., Ltd, demonstrated that immunization of monkeys (n = 3) with mPEG-KLH induced anti-PEG IgG that was exclusively directed against the PEG backbone, not the terminal methoxy group.
Background
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). To aid research in this important area, we have developed a anti-PEG ELISA kit. Used for new drug research and development
Monkey Anti-PEG IgG ELISA kit (MAK-0014).pdf
