ELISA Assay Kit for Anti-HER-2 h-mAb in Human Serum

Materials Provided

Cat. No. Components Size (480 tests)
PK401-213 Human HER-2 10 μg
PK402-213 Biotinylated anti-HER-2 antibody 10 μg
PK403-213 Streptavidin-HRP 10 μg
PK404-213 Anti-HER-2 antibody 30 μg

Background

How a therapeutic antibody is metabolized in the body is pertinently relevant to its efficacy. Therefore, pharmacokinetics study is an important part of the drug development.

HER-2 is probably the most targeted molecule in today’s pharmaceutical industry, thanking to the clinical success of Herceptin®. Many investigational anti-HER-2 mAbs are being developed. There is a growing need for a standard assay that can be used to facilitate the study of their pharmacokinetics.

Assay Principles

The enclosed ELISA assay kit for anti-HER-2 h-mAb in human serum is based on an enzyme immunoassay (ELISA) between recombinant HER-2 protein (ECD) and a biotinylated anti-HER-2 monoclonal antibody. The method employs the principle of competitive ELISA, allowing quantification of monoclonal anti-HER-2 antibodies in human serum.

The assay only involves the following steps:

  1. Coat the plate with human HER-2;
  2. Add the mixture of your sample and biotinylated anti-HER-2 antibody;
  3. Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate;
  4. Record the OD readings and analyze the serum concentration.

Reconstitution

See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Advantages

  • Wide Coverage - There is no requirement for the species and subtype of the antibody.

  • Easy Process - There is no longer a need to generate anti-drug antibodies for PK studies.

  • Clean Result - The use of the biotinylated antibody in the competitive ELISA significantly alleviates the issue of background noise issue that often occurs in traditional ELISA method.

Storage

Upon receipt, please store all items at -20 ℃. After reconstitution, the stock solution should be kept at -80 ℃. It is recommend not to freeze  thaw more than 3 times.  No activity loss is observed after storage at:

  • Room temperature (RT) for 1 month in lyophilized state;
  • -20 ℃ for 6 months in lyophilized state;
  • -80 ℃ for 4 months under sterile conditions after reconstitution.

Typical Data

Inhibition of HER-2: biotinylated anti-HER-2 antibody binding by an anti-HER-2 antibody.

Serial dilutions of the Therapeutic Anti-HER-2 Antibody (1:2 serial dilutions, from 20 μg/mL to 0.039 μg/mL) was added into HER-2: Biotinylated Anti-HER-2 Antibody binding reactions.