EC50 Determination Service for Antibody Drugs

Principle of the assay

This EC50 determination for antibody employs an indirect ELISA method. In this assay, standard 96-well plates (12 strips with 8 wells/strip) are coated with a specific protein, which combines with the corresponding antibody present in a sample, which is diluted in a series of dilution fold for EC50 determination. The wells are washed, and biotinylated detection antibody such as anti-human IgG antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells, and color develops in proportion to the amount of the specific protein antibody bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. The EC50 concentration can be calculated by plot the series dilution antibody or a sample as 4-PL (4 parametric logistic) standard curve, with standard concentration on the x-axis and absorbance on the y-axis.

Service Features

  • High quality analyses performed
  • Just send your antibody/sample, will send you EC50 tested results
  • Affordable
  • Rapid turn-around
  • Bulk discounts
  • Long-term service contracts available

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What's Included

  • Standard curve with tested EC50 concentration (see below typical data) in Excel
  • Standard curve raw data

Applications of testing service for EC50 of Antibody Determination

  • Pharmacokinetics and concentration of antibody such as response relationship
  • Immunogenicity of antibody
  • Evaluating the effect of antibody on pharmacokinetic and safety profiles

Typical Data

Typical data figure 1
Fig 1. Panitumumab antibody was tested for determining EC50 with EGFR protein binding. A series dilution of Panitumumab antibody from 0 - 100 ng/ml was prepared and performed the assay.
Typical data figure 2
Fig 2. Atezolizumab antibody was tested for determining EC50 with PD-L1 protein binding. A series dilution of Panitumumab antibody from 0 - 15 ng/ml was prepared and performed the assay.