Cat. No.
MAK-0008
Product Name
Sandwich ELISA for detection of PEGylated proteins kit
Storage
2–8°C.
Principle
Sandwich ELISA
Intended Use
This kit is for research use only. Under no circumstances should it be used for therapeutic or diagnostic applications.
Principle Of The Assay
The assay uses two different monoclonal antibodies that recognize the PEG backbone. One is coated on wells of the microtiter plate and is used for capture; the other is conjugated to horseradish peroxidase (HRP) and is used for detection. Serum samples are diluted at least 10-fold in the provided dilution buffer and incubated alongside reference standards1 in the microtiter wells for 45-minutes. The wells are subsequently washed. HRP conjugate is added and incubated for 45 minutes. If present, PEG molecules are sandwiched between the capture and detection antibodies. The wells are then washed to remove unbound HRP-conjugate. TMB is added and incubated for 20 minutes. This results in the development of a blue color. Color development is stopped by the addition of Stop Solution, changing the color to yellow. Absorbance is measured at 450 nm. The concentrations of PEG in the samples are derived from a standard curve.
Background
PEGylation of biologics prolongs their half-life by slowing proteolytic degradation and decreasing the rate of clearance from the circulatory system. The pharmacodynamics of PEGylated proteins are often evaluated using specific assays for the protein itself. That approach often requires the time consuming and expensive construction of an ELISA for the protein of interest. The PEG ELISA manufactured by Mabioway Co., Ltd. allows measurement of the PEG portion of the PEGylated protein and is therefore suitable for assessment of the pharmacodynamics of a range of PEGylated proteins.
Sandwich ELISA for detection of PEGylated proteins kit (MAK-0008).pdf
