Heterophile Antibody Interference
This phenomenon occurs when endogenous antibodies, typically with broad and nonspecific affinity for various antigens, including animal-derived immunoglobulins, disrupt immunoassay function. Due to their bivalent nature, these antibodies can inadvertently bridge the capture and detection antibodies in immunometric assays even in the absence of the target analyte, producing falsely elevated results.
Most individuals have only low titers of heterophile antibodies, and modern immunoassays include blocking agents to neutralize their effect. However, in certain individuals with high titers, such as patients exposed to therapeutic monoclonal antibodies or animal-derived immunoglobulins, these blockers may be insufficient, leading to spurious assay results. In some cases, the source of interference is identifiable; in others, it remains clinically unpredictable.
Use of Single-Chain Antibodies (scFv): Employing single-chain antibodies can reduce heterophilic antibody interference, which can contribute to the hook effect. Single-chain antibodies have a smaller size and reduced likelihood of cross-linking, thereby improving assay reliability.
Illustration: Heterophile Antibody Interference
References
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Haddad, R.A., Giacherio, D. & Barkan, A.L. Interpretation of common endocrine laboratory tests: technical pitfalls, their mechanisms and practical considerations. Clin Diabetes Endocrinol 5, 12 (2019). https://doi.org/10.1186/s40842-019-0086-7
Levinson SS, Miller JJ. Towards a better understanding of heterophile (and the like) antibody interference with modern immunoassays. Clin Chim Acta. 2002 Nov;325(1-2):1-15. doi: 10.1016/s0009-8981(02)00275-9. PMID: 12367762.
Bolstad N, Warren DJ, Bjerner J, Kravdal G, Schwettmann L, Olsen KH, Rustad P, Nustad K. Heterophilic antibody interference in commercial immunoassays; a screening study using paired native and pre-blocked sera. Clin Chem Lab Med. 2011 Sep 8;49(12):2001-6. doi: 10.1515/CCLM.2011.702. Erratum in: Clin Chem Lab Med. 2012 Feb;50(2):409. PMID: 21899496.
Cheng X, Guo X, Chai X, Hu Y, Lian X, Zhang G. Heterophilic antibody interference with TSH measurement on different immunoassay platforms. Clin Chim Acta. 2021 Jan;512:63-65. doi: 10.1016/j.cca.2020.11.018. Epub 2020 Dec 5. PMID: 33285118.
Wheeler, Michael J. Hormone Assays in Biological Fluids. Edited by Michael J. Wheeler, 2nd ed. 2013., vol. 1065, Humana Press, 2013, https://doi.org/10.1007/978-1-62703-616-0.
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