Multiplex cytokine analyses in patients with rheumatoid arthritis require use of agents blocking heterophilic antibody activity

Abstract

When analysing sera from patients with RA with multiplex bead ELISA, the assay should be evaluated for interference by heterophilic antibodies, and if present corrected with, for example, HeteroBlock.

Rheumatoid factor and falsely elevated results in commercial immunoassays: data from an early arthritis cohort

“…In the 27-plex cytokine assay, our study revealed interference in 70% of tested sera, and 17 out of 27 cytokine assays. In line with other publications, our results showed that interference from RF can be reduced by addition of blocking agents [ 10 – 12 ]. We encourage the assay producer to protect these assays against interference from RF, e.g.…”

“…However, levels of RF IgA and IgM were only weakly correlated with the level of anti-animal IgG reactivity in our study, suggesting that antibody properties such as avidity and specificity could be equally important. In line with our results, other studies have shown that interference in multiplex cytokine assays is unpredictable in relation to RF level, and all sera from RA patients should be treated as likely to interfere in multiplex assays [ 10 , 12 ].…”

Rheumatoid factor and falsely elevated results in commercial immunoassays: data from an early arthritis cohort

“…In the 27-plex cytokine assay, our study revealed interference in 70% of tested sera, and 17 out of 27 cytokine assays. In line with other publications, our results showed that interference from RF can be reduced by addition of blocking agents [ 10 – 12 ]. We encourage the assay producer to protect these assays against interference from RF, e.g.…”

“…However, levels of RF IgA and IgM were only weakly correlated with the level of anti-animal IgG reactivity in our study, suggesting that antibody properties such as avidity and specificity could be equally important. In line with our results, other studies have shown that interference in multiplex cytokine assays is unpredictable in relation to RF level, and all sera from RA patients should be treated as likely to interfere in multiplex assays [ 10 , 12 ].…”

Heterophilic Interference of Rheumatoid Factor in TSH Immunometric Assay: A Cross-Sectional Observational Study

“…Additionally, other studies of RF interference with multiplex cytokine assays have shown that the relationship between RF level and interference is unpredictable. [ 16 30 31 ] Importantly, corrected TSH (based on the mean of dilutions or post-HB treated TSH) did not change the clinical interpretation in most of the concerned cases. This indicates that RF is a weaker heterophile antibody causing a lesser degree of interference and having minimal effect on the interpretation of TSH assay results by modern immunoassay platforms.…”

Heightened Levels of Antimicrobial Response Factors in Patients With Rheumatoid Arthritis

“…Autoantibodies produced in RA such as RF cause interference in some immunoassays (91). RF can generate false signals by bridging capture and detection antibodies in sandwich ELISAs (92), an effect that can be mitigated through HeteroBlock (91,93). A lack of corrective measures to block the RF interference in the study reported by Wen et al (90) could explain their results.…”

Rheumatoid factor and falsely elevated results in commercial immunoassays: data from an early arthritis cohort

“…In the 27-plex cytokine assay, our study revealed interference in 70% of tested sera, and 17 out of 27 cytokine assays. In line with other publications, our results showed that interference from RF can be reduced by addition of blocking agents [ 10 – 12 ]. We encourage the assay producer to protect these assays against interference from RF, e.g.…”

“…However, levels of RF IgA and IgM were only weakly correlated with the level of anti-animal IgG reactivity in our study, suggesting that antibody properties such as avidity and specificity could be equally important. In line with our results, other studies have shown that interference in multiplex cytokine assays is unpredictable in relation to RF level, and all sera from RA patients should be treated as likely to interfere in multiplex assays [ 10 , 12 ].…”

Heterophilic Interference of Rheumatoid Factor in TSH Immunometric Assay: A Cross-Sectional Observational Study

“…Additionally, other studies of RF interference with multiplex cytokine assays have shown that the relationship between RF level and interference is unpredictable. [ 16 30 31 ] Importantly, corrected TSH (based on the mean of dilutions or post-HB treated TSH) did not change the clinical interpretation in most of the concerned cases. This indicates that RF is a weaker heterophile antibody causing a lesser degree of interference and having minimal effect on the interpretation of TSH assay results by modern immunoassay platforms.…”

Heightened Levels of Antimicrobial Response Factors in Patients With Rheumatoid Arthritis

“…Autoantibodies produced in RA such as RF cause interference in some immunoassays (91). RF can generate false signals by bridging capture and detection antibodies in sandwich ELISAs (92), an effect that can be mitigated through HeteroBlock (91,93). A lack of corrective measures to block the RF interference in the study reported by Wen et al (90) could explain their results.…”

Rheumatoid factor and falsely elevated results in commercial immunoassays: data from an early arthritis cohort

“…In the 27-plex cytokine assay, our study revealed interference in 70% of tested sera, and 17 out of 27 cytokine assays. In line with other publications, our results showed that interference from RF can be reduced by addition of blocking agents [ 10 – 12 ]. We encourage the assay producer to protect these assays against interference from RF, e.g.…”

“…However, levels of RF IgA and IgM were only weakly correlated with the level of anti-animal IgG reactivity in our study, suggesting that antibody properties such as avidity and specificity could be equally important. In line with our results, other studies have shown that interference in multiplex cytokine assays is unpredictable in relation to RF level, and all sera from RA patients should be treated as likely to interfere in multiplex assays [ 10 , 12 ].…”

Heterophilic Interference of Rheumatoid Factor in TSH Immunometric Assay: A Cross-Sectional Observational Study

“…Additionally, other studies of RF interference with multiplex cytokine assays have shown that the relationship between RF level and interference is unpredictable. [ 16 30 31 ] Importantly, corrected TSH (based on the mean of dilutions or post-HB treated TSH) did not change the clinical interpretation in most of the concerned cases. This indicates that RF is a weaker heterophile antibody causing a lesser degree of interference and having minimal effect on the interpretation of TSH assay results by modern immunoassay platforms.…”

Heightened Levels of Antimicrobial Response Factors in Patients With Rheumatoid Arthritis

“…Autoantibodies produced in RA such as RF cause interference in some immunoassays (91). RF can generate false signals by bridging capture and detection antibodies in sandwich ELISAs (92), an effect that can be mitigated through HeteroBlock (91,93). A lack of corrective measures to block the RF interference in the study reported by Wen et al (90) could explain their results.…”