Avian antibodies can eliminate interference due to complement activation in ELISA

Area of Science:

• Immunology
• Biochemistry

Background:

• Mammalian antibodies can activate the human complement system.
• Activated complement components can block antibody binding sites, leading to inaccurate assay results.
• Serum samples contain an active complement system that is inactivated during storage.

Purpose of the Study:

• To evaluate the suitability of chicken antibodies in immunological assays compared to mammalian antibodies.
• To investigate the interference caused by complement system activation in immunoassays.
• To demonstrate how chicken antibodies can overcome complement-mediated interference.

Main Methods:

• Comparison of absorbance values using goat (mammalian) and chicken antibodies in immunological assays.
• Assessment of complement system activity in serum samples.
• Analysis of antibody binding epitope accessibility.

Main Results:

• Active complement in undiluted samples reduced absorbance values by approximately 50% with goat antibodies.
• Chicken antibodies did not show this reduction in absorbance, indicating no complement activation.
• This interference leads to variable and erroneous test results depending on sample handling.

Conclusions:

• Chicken antibodies do not activate the human complement system.
• Using chicken antibodies in immunoassays eliminates complement-mediated interference, improving test result reliability.
• Egg yolk-derived antibodies offer advantages for designing robust and accurate solid-phase immunometric assays.