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Immunometric assay interference: incidence and prevention.

Johan Bjerner¹, Kjell Nustad, Lars F Norum

¹Central Laboratory, Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway. johan.bjerner@klinmed.uio.no

Clinical Chemistry

|March 20, 2002

Summary

Reducing interference in immunometric assays is crucial. Heat-treated murine immunoglobulin or Fc fragment removal significantly improved assay performance by minimizing interference.

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Area of Science:

Immunology
Biochemistry
Analytical Chemistry

Background:

Interference in immunoassays can lead to inaccurate results.
Two-site, two-step immunometric assays are widely used but susceptible to interference.
Minimizing interference is essential for reliable diagnostic testing.

Purpose of the Study:

To reduce interference in an in-house two-site, two-step immunometric assay.
To evaluate the effectiveness of heat-treated nonspecific murine immunoglobulin and Fc fragment removal in mitigating assay interference.

Main Methods:

Tested 11,261 samples using a carcinoembryonic antigen (CEA) assay.
Compared assays with and without bovine immunoglobulin, with added murine immunoglobulin (MAK33), and with Fc fragments removed from the capture antibody.

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Assessed interference frequency and statistical significance across different assay modifications.

Main Results:

Initial interference frequency was 4.0%.
Addition of heat-treated MAK33 significantly reduced interference to 0.86% (15 mg/L) and 0.06% (50 mg/L).
Fc fragment removal reduced interference to 0.10%; no significant differences in interference based on age or gender were observed.

Conclusions:

Heat-treated nonspecific murine immunoglobulin in the buffer effectively reduces interference.
Removal of the Fc fragment from the capture antibody also improves assay performance.
These methods enhance the reliability of two-site, two-step immunometric assays utilizing mouse monoclonal antibodies.