Cat. No.
MAK-0015
Product Name
Monkey Anti-PEG IgM 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 IgM monoclonal antibody (LDI clone 2C11-1-51) for detection. Serum or plasma samples are diluted and incubated alongside standards in the microtiter wells for 1 hour. The wells are subsequently washed, and HRP conjugate is added and incubated for 45 minutes. Anti-PEG IgM molecules are thus sandwiched between immobilized PEG and the detection antibody conjugate. The wells are then washed to remove unbound HRP-labeled antibodies. 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. Optical density is measured spectrophotometrically at 450 nm. The concentration of anti-PEG IgM 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, in mice, rats and rabbits have demonstrated that most anti-PEG antibodies induced by immunization with PEGylated proteins are directed against the PEG backbone.
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 IgM ELISA kit (MAK-0015).pdf
