PG, ELISA, (RUO) Research Use Only

The PG-ELISA is a solid phase Enzyme Amplified Sensitivity Immunoassay (ELISA) performed on microtiter plate. The assay is based on an oligoclonal system in which a blend of monoclonal antibodies (MAbs) directed against distinct epitopes of PG are used. Antibody-producing cells are immortilized using the myeloma cell fusion method of Kohler and Milstein. A hybridoma cell is produced which secretes specific homogeneous antibodies. The use of a number of distinct MAbs avoids hyperspecificity and allows highly sensitive assays with extended standard range and short incubation time. Standards or samples containing PG react with capture monoclonal antibodies (MAbs 1) coated on the microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP).

After an incubation period allowing the formation of a sandwich : coated MAbs 1 - PG - MAb 2 - HRP, the microtiter plate is washed to remove unbound enzyme labelled antibodies. Bound enzyme-labelled antibodies are measured through a chromogenic reaction. Chromogenic Solution (TMB+H2O2) is added and incubated.

The reaction is stopped with the addition of Stop Solution (HCl) and the microtiter plate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance which is proportional to the PG concentration.

A standard curve is plotted and PG concentrations in a sample is determined by interpolation from the standard curve.