High Sensitivity Polyethylene Glycol (PEG) ELISA Kit

Sample

Serum, plasma

Species Reactivity

N/A

Detection Method

Competitive-ELISA

Intended Use

The kit can be used for the detection of proteins containing one or more PEG chains as well as unconjugated PEG.

Contents of Kit

No.	Components	Size	Storage Conditions
1	PEG Coated 96-well Microtiter Plate	12 × 8-wells	Store at -20°C
2	HRP anti-PEG Conjugate	1 vial	Store at -20°C.
3	HRP PEG Diluent	50 ml	2-8°C
4	PEG-BSA Standard	1 vial	Store at -20°C.
5	HRP PEG Wash Buffer	50 ml	2-8°C
6	TMB Reagent	11 ml	2-8°C
7	Stop Solution	11 ml	2-8°C

Storage

Upon receipt the HRP Anti-PEG vial, the PEG BSA standard and the PEG coated 96 well plate should be placed in a -20°C freezer until use. Do not store at lower temperatures. The remainder of the kit should be stored in a refrigerator at 2-8°C. The microtiter strips should be kept in a sealed bag with desiccant to minimize exposure to damp air. Test kits will remain stable for six months from the date of purchase provided that the components are stored as described above.
It is important that the plate, diluent and TMB reagent equilibrate to room temperature before use.

Sensitivity

Characteristics of PEGylated BSA and free PEG in the High Sensitivity PEG ELISA. Please note that the concentrations of PEGylated BSA refer only to the polypeptide content and do not include mass contributed by PEG. EC50 values represent mean +/- SD from 3-6 studies.

General Description

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 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.

Standard Curve

A representative standard curve with optical density readings at 450 nm on the Y axis against BSA-mPEG 20 kDa concentration on the X axis is shown below. Please note that the X-axis is a log₁₀ scale.