Detection and Elimination of Interference by the Heterophilic Antibody in Antibody Microarray–Based Immunoassay

False‐positive semiquantitative immunochromatography assays for procalcitonin in three patients with rheumatoid arthritis—A case series

“…2,6 When two monoclonal mouse antibodies are used in a single assay, the frequency of false-positive results increases substantially vs use of a mouse monoclonal antibody and a polyclonal antibody from another species. 7 Heterophilic antibody interference has been reported for many tumor markers (alpha-fetoprotein, CA125, beta-human chorionic gonadotropin, squamous cell carcinoma antigen, and prostate-specific antigen), hormones (thyrotrophic hormone and estradiol), and infection markers (influenza and human hepatitis B antigen) using immunochromatography assays. 6,[8][9][10] Commercial quantitative methods have been improved to reduce heterophilic interference by removing the Fc portion and avoid the binding of heterophilic antibodies; this can also be achieved by adding a HAMA blocker or adsorbing the heterophilic antibodies using IgG from the same animal or serum, as a secondary antibody.…”

“…The mechanism underlying the false‐positive caused by HAMA and other heterophilic antibodies is thought to be their binding to, or cross‐reaction with, the antikatacalcin antibody labeled with gold colloid (mouse monoclonal antibody) and antihuman calcitonin antibody (sheep polyclonal antibody) on the test line 2,6 . When two monoclonal mouse antibodies are used in a single assay, the frequency of false‐positive results increases substantially vs use of a mouse monoclonal antibody and a polyclonal antibody from another species 7 . Heterophilic antibody interference has been reported for many tumor markers (alpha‐fetoprotein, CA125, beta‐human chorionic gonadotropin, squamous cell carcinoma antigen, and prostate‐specific antigen), hormones (thyrotrophic hormone and estradiol), and infection markers (influenza and human hepatitis B antigen) using immunochromatography assays 6,8–10 .…”