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Antigen attachment in ELISA

O P Lehtonen, M K Viljanen

Journal of Immunological Methods

|January 1, 1980

Summary

Antigen leakage from solid phases in enzyme-linked immunosorbent assays (ELISA) can cause errors. Covalent coupling of antigen to activated paper minimizes this leakage, making it suitable for standardizing ELISA methods.

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Area of Science:

Immunology
Biochemistry
Analytical Chemistry

Background:

Enzyme-linked immunosorbent assays (ELISA) are widely used for antibody detection.
The stability of the antigen immobilized on the solid phase is crucial for assay reliability.
Antigen detachment from the solid phase can lead to inaccurate results in antibody quantification.

Purpose of the Study:

To investigate antigen leakage during different stages of ELISA.
To compare antigen stability on various solid phase materials: polystyrene, nylon, and cyanogen bromide activated paper.
To evaluate the suitability of activated paper as a reference method for ELISA standardization.

Main Methods:

Chicken anti-bovine serum albumin (BSA) antibodies were used in the ELISA.
Antigen immobilization and subsequent leakage were monitored on polystyrene, nylon, and activated paper solid phases.
Antigen detachment was assessed during sample incubation, antibody incubation, and washing steps.

Main Results:

Significant antigen leakage occurred from polystyrene (30%) and nylon (60%) solid phases.
Antigen detachment was observed throughout all assay steps, including incubation and washing.
Covalent coupling of antigen to activated paper resulted in significantly less batch-to-batch variation and antigen detachment compared to plastics.

Conclusions:

Antigen leakage from polystyrene and nylon solid phases can compromise ELISA accuracy.
Activated paper, utilizing covalent antigen coupling, offers superior antigen stability.
Activated paper is a promising solid phase for developing standardized and reliable ELISA systems.

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