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Hook effect

The Hook effect, also known as the high-dose hook effect or postzone phenomenon, is an interference in sandwich immunoassays where excessively high concentrations of the target result in falsely low measured values due to saturation of assay antibodies. This phenomenon arises primarily in one-step heterogeneous immunoassays, where the excess binds independently to both capture and detection antibodies, inhibiting the formation of the antibody--antibody complex required for signal detection. The name derives from the characteristic "hooked" shape of the dose-response curve, which rises with increasing concentration, plateaus, and then declines at very high levels. The Hook effect was first described in by Miles et al. in a two-site immunoradiometric for , where high levels led to unexpectedly low signals due to saturation. The term "hook effect" was later adopted to describe the curved appearance of the dose-response plot, distinguishing it from the earlier prozone effect observed in reactions since the early . The effect is distinct from the prozone effect (antibody excess) but shares similarities in causing assay nonlinearity. It is most pronounced in assays without built-in dilution steps or kinetic monitoring, such as certain enzyme-linked immunosorbent assays (ELISAs) and lateral flow immunoassays used in point-of-care testing. Clinically, the Hook effect can lead to underestimation of critical biomarkers, potentially resulting in missed diagnoses, such as in assays for prolactin, thyroid-stimulating hormone (TSH), human chorionic gonadotropin (hCG), and tumor markers like prostate-specific antigen (PSA). In pregnancy testing, a variant can occur due to hCG beta-core fragments, affecting up to 18% of point-of-care devices and causing false negatives in conditions like ectopic or molar pregnancies. Mitigation strategies include serial sample dilutions and real-time kinetic monitoring to ensure accurate results in high-stakes scenarios.

Introduction

Definition

The hook effect, also known as the high-dose hook effect, is an analytical interference in immunoassays where excessively high concentrations produce falsely low or negative results, paradoxically decreasing the measured signal as analyte levels rise beyond a certain . This phenomenon arises in assays designed to detect large molecules with multiple epitopes, leading to an underestimation that can mimic low analyte presence despite actual abundance. In a sandwich immunoassay, the process begins with a capture immobilized on a that binds to one on the target . A detection , often labeled with an or fluorescent tag, then binds to a distinct on the same , forming an antibody-- "sandwich" complex; the bound label generates a quantifiable signal proportional to concentration in typical ranges. Under normal conditions, signal intensity increases linearly with concentration, but the hook effect disrupts this by causing signal saturation and decline at extreme levels, resulting in a characteristic hook-shaped dose-response curve rather than a straightforward linear or sigmoidal pattern. This interference primarily affects assays for macromolecular analytes such as hormones like (hCG) and , as well as tumor markers including (AFP) and cancer antigen 125 (CA-125). The hook effect is a specific instance of the postzone (antigen excess) in sandwich immunoassays, distinct from the prozone phenomenon (antibody excess), though both cause similar interferences; these are explored in greater detail in the Mechanism section.

Historical Context

The prozone phenomenon, a related involving antibody excess and an early concept in , was first recognized in the early during the development of serological tests for , particularly in and reactions like the introduced in 1906. In these assays, excessive antibody concentrations prevented visible lattice formation between antigen and antibody, leading to false-negative results despite the presence of . This observation highlighted the limitations of qualitative immunological tests reliant on visible precipitates or clumps. In the 1930s, researchers such as John R. Marrack advanced the understanding through quantitative studies of precipitin reactions, demonstrating characteristic curves where precipitation peaked at equivalence and declined in zones of or antigen excess. Marrack's work, including analyses of precipitate composition in antigen- mixtures, linked these zones explicitly to excess reactants, providing a foundational framework for interpreting such anomalies in classical . By the and , this linkage to antibody excess was solidified in broader immunological reactions, influencing the design of more sensitive assays. The 1970s marked the application of the phenomenon to modern quantitative immunoassays, with the first explicit description of the "hook effect" in two-site immunoradiometric assays (IRMA) for ferritin by Miles et al. in 1974. Pioneers like Rosalyn Yalow, who developed radioimmunoassay (RIA) techniques in the 1950s and received the Nobel Prize in 1977 for their contributions to immunoassay methodology, indirectly facilitated this transition by enabling precise measurement of analytes, though RIA's competitive format was less prone to the effect. By the 1980s and 1990s, the hook effect was identified in enzyme-linked immunosorbent assays (ELISA) and emerging chemiluminescent assays, where high analyte levels saturated capture and detection antibodies in sandwich formats. Terminology evolved from the classical "prozone" (antibody excess) and "postzone" (antigen excess) used in early 20th-century serology to "hook effect" by the 1980s, reflecting the curved, hook-like shape of dose-response plots in automated, quantitative systems. This shift emphasized the practical implications in clinical diagnostics, distinguishing it from qualitative observations. As of 2025, the hook effect remains relevant in modern point-of-care , with recent research focusing on mitigation strategies in lateral flow devices.

Mechanism

Prozone Phenomenon

The prozone phenomenon, distinct from but related to the hook effect, refers to the antibody-excess scenario where an overabundance of antibodies relative to antigens disrupts the formation of detectable immune complexes in serological assays. In this condition, excess soluble antibodies bind to limited antigens, forming primarily small, monovalent complexes that remain soluble and fail to aggregate into the cross-linked lattices necessary for precipitation or signal amplification in detection systems. This leads to falsely low or negative results, as the soluble complexes do not precipitate or generate measurable signals. The underlying molecular interactions are governed by the lattice hypothesis, first proposed by Marrack in 1934, which describes how multivalent antigens and bivalent antibodies must interact in proportional ratios to form an extended network for visible precipitation. In prozone conditions, antigens are rapidly saturated by excess antibodies through monovalent binding, blocking the bivalent attachment sites required for cross-linking and lattice expansion; consequently, only non-aggregating Ab-Ag complexes form, preventing the multivalent structures essential for detection. This can be conceptually represented by the for monovalent binding: \text{Ab} + \text{Ag} \rightleftharpoons \text{AbAg} in contrast to the lattice-forming multivalent complex: (\text{AbAg})_n \quad (n > 1) where the latter enables precipitation through repeated cross-links.[](Marrack JR. The Chemistry of Antigens and Antibodies. Medical Research Council Special Report Series No. 194. London: HMSO; 1934.) The prozone phenomenon typically arises in environments with high antibody concentrations, such as undiluted sera from hyperimmune states or early phases of polyclonal antibody responses where titers are disproportionately elevated relative to antigen levels. It is most prevalent in classical precipitation and agglutination tests, where antibody amounts are not precisely controlled, but is rare in contemporary one-step sandwich immunoassays due to fixed antibody immobilization and optimized reagent ratios that minimize excess.[](Reverberi R, Reverberi L. Factors affecting the antigen-antibody reaction. Blood Transfus. 2007;5(4):227-240. doi:10.2450/2007.0047-07.)[](van Oss CJ. Nature and role of cross-reactivity. J Immunoassay Immunochem. 2000;21(3):209-219. doi:10.1080/01971520009349532.) Quantitatively, the dose-response curve under prozone conditions shows a precipitous signal decline when antibody levels surpass the zone by 10- to 100-fold, reflecting the saturation threshold beyond which lattice formation collapses.[](Henry JB, ed. Clinical Diagnosis and Management by Laboratory Methods. 20th ed. Saunders; 2001.) This non-linear response underscores the need for dilution protocols in high-titer samples to restore and accurate detection.

Postzone Phenomenon

The postzone phenomenon, also known as the high-dose hook effect in its antigen-excess form, occurs in immunoassays when excessively high concentrations of the target () lead to falsely low or undetectable signals. In this scenario, the surplus saturates the capture antibodies immobilized on the solid phase and the detection antibodies (often labeled) independently, forming incomplete complexes such as antibody- (Ab-Ag) and -antibody (Ag-Ab) pairs that fail to assemble into the full detectable structure (Ab-Ag-Ab). This saturation prevents the formation of the ternary complex required for signal generation, resulting in a paradoxical decrease in measured concentration despite its abundance. At the molecular level, the excess competes for binding sites on both types, blocking the linkage needed for formation and leading to an accumulation of free or singly bound that does not contribute to the signal. The detectable signal is proportional to the concentration of the Ab-Ag-Ab complex, following that peak at optimal levels and decline sharply when concentration greatly exceeds availability ([Ag] >> [Ab]), often described by a bell-shaped dose-response curve. This arises because the high density promotes the formation of higher-order complexes or multimeric aggregates that deplete available detection antibodies without enhancing the test signal. The postzone effect is particularly prevalent in one-step immunoassays, where capture and detection antibodies are incubated simultaneously with the sample, allowing immediate without intermediate washing steps to remove excess ; it is common for s at concentrations exceeding 1,000 times the assay's upper limit of quantification, such as certain tumor markers or inflammatory proteins. This issue is exacerbated in automated high-throughput systems lacking dilution protocols or signal verification checks, as the excess antigen can overwhelm the limited antibody binding sites (typically in the nanomolar range) before favors formation. Unlike the prozone phenomenon, which involves antibody excess, the postzone specifically disrupts assays through analyte overload in modern formats. Quantitatively, the hook curve's —where signal begins to decline—typically occurs at levels 10³ to 10⁶ times the standard range, depending on affinity and design; for example, in lateral flow assays for allergens, signals peak around 10–100 before dropping at >250 . Signal recovery upon (e.g., 1:10 or 1:100) is a hallmark, as reducing concentration below levels restores proportional sandwich formation and yields results that, when multiplied by the dilution factor, reveal the true high value. This behavior underscores the need for awareness in interpreting non-linear responses in antigen-rich samples.

Laboratory and Clinical Importance

In Vitro Effects in Immunoassays

The hook effect manifests in vitro primarily within sandwich immunoassays, where excessively high analyte concentrations lead to antibody saturation and falsely low signal outputs, mimicking normal or reduced levels despite true elevation. This artifact arises in one-step formats, such as enzyme-linked immunosorbent assays (), chemiluminescent immunoassays (CLIA), and electrochemiluminescent immunoassays (ECLIA), where capture and detection antibodies bind the analyte simultaneously without intermediate washing steps. In contrast, competitive assays or two-step protocols with separation phases are generally resistant, as they allow excess analyte removal before detection. Vulnerable analytes include tumor markers and hormones, often yielding false-low results at concentrations far exceeding assay upper limits. For (PSA), hook effect has been documented in metastatic cases, with reported values as low as normal ranges despite very high actual levels, as seen in one-step immunometric assays. Similarly, (AFP) in hepatoblastoma patients can trigger the effect at very high concentrations in certain automated systems, leading to underestimation and potential misclassification as benign conditions. In hormone assays, (hCG) during gestational trophoblastic disease or molar pregnancies may report as <10,000 IU/L when true concentrations surpass 500,000 IU/L, such as one case with 5,899,478 mIU/mL post-dilution confirmation. Severity of the hook effect varies based on assay design and analyte properties. Larger analytes or those with multimeric structures exacerbate saturation due to inefficient binding kinetics, while lower antibody affinity accelerates the onset at lower excess thresholds. Shorter incubation times in rapid one-step formats heighten risk by limiting equilibrium, and instrument-specific limits—often triggering at 10- to 100-fold above the reportable range—determine detection thresholds, as in prolactin assays hooking at >10,000 ng/mL. These factors collectively influence the of the . Diagnostic consequences include underdiagnosis of severe pathologies, as falsely normal results delay interventions. In ectopic or molar pregnancies, undetected high hCG risks rupture or progression without prompt monitoring. For malignancies, such as macroprolactinomas or carcinomas, underestimated tumor markers like (>26,000 ng/mL actual vs. 164 ng/mL reported) or (>200,000 ng/mL actual) may lead to inappropriate or surgical errors. Overall, this artifact underscores the need for vigilance in high-suspicion cases to avoid clinical missteps.

In Vivo Observations

Clinical observations of the hook effect arise in patients with physiological conditions involving excessively high concentrations of circulating or in their samples, leading to falsely low measurements in diagnostic . In diseases involving massive antigen secretion, such as certain tumors, or elevated antibody levels in autoimmune disorders with high IgG, these high levels can cause the antigen or antibody surplus that results in hook effect artifacts during clinical testing. For instance, in macroprolactinomas—pituitary tumors secreting large amounts of —the hook effect arises when prolactin levels overwhelm immunoassay antibodies, resulting in artifactually normal readings despite hyperprolactinemia. Similarly, in autoimmune conditions like those with anti-glutamic acid decarboxylase (GAD) antibodies, extreme elevations can saturate capture sites, producing misleadingly low results. Documented clinical cases highlight the diagnostic challenges posed by the hook effect, particularly in and . Reports from the 1980s described false-normal prolactin levels in patients with macroprolactinomas, where undiluted serum assays underestimated concentrations by orders of magnitude, delaying recognition of invasive tumors. In , a gestational trophoblastic tumor, high beta-human chorionic gonadotropin (beta-hCG) levels have caused hook effect-induced false lows in immunoassays, contributing to missed diagnoses and treatment delays; a 1990 case illustrated this when beta-hCG appeared at only 283 mIU/L despite actual levels exceeding 1,000,000 mIU/L, complicating ovarian tumor management. Post-2000 studies on (GTD) further document similar issues, such as a 2015 report of hook effect delaying GTD diagnosis at beta-hCG >500,000 IU/L and a 2022 emergency case where undiagnosed presented with negative urine tests due to antigen excess. Unlike in vitro scenarios, clinical hook effects occur without prior sample dilution, allowing physiological concentrations to directly impact assay performance during testing, often exacerbated by pre-analytical handling like incomplete mixing or delayed processing. Literature estimates incidence at approximately 1-5% in high-burden diseases, such as macroprolactinomas or advanced GTD, where antigen loads routinely surpass assay thresholds. Case reports spanning the to consistently demonstrate resolution upon post-collection dilution, revealing true elevations that alter patient management; for example, 1990s prolactinoma cases showed 10- to 100-fold increases post-dilution, prompting surgical or therapeutic interventions. These discrepancies carry significant implications for tumor staging—potentially underestimating disease extent in —and hormone therapy decisions, such as avoiding unnecessary agonists in misdiagnosed prolactinomas.

Detection and Mitigation Strategies

Identifying the Hook Effect

The primary method for identifying the hook effect in suspect samples involves of the specimen, typically at ratios such as 1:10, 1:100, and 1:1000, followed by retesting on the platform. If the hook effect is present, the measured concentration will paradoxically increase after dilution and then align with the expected dose-response curve at higher dilutions, confirming the due to analyte excess. This approach is straightforward and widely recommended for verification, as it directly addresses the saturation of capture antibodies in immunoassays. Secondary indicators of the hook effect include discordant laboratory results, where the yields a low or negative value despite strong clinical suspicion of elevated levels, such as in cases of suspected elevation or with low hCG readings. Additional checks involve assessing non-linear behavior in curves or performing experiments, in which post-dilution exceeds 100% of the expected value based on the undiluted result, signaling potential . These indicators prompt further but require confirmation through dilution studies to rule out other interferences. Modern analyzers incorporate features to aid in hook effect detection, such as built-in algorithms that monitor raw signal intensities or automatically flag results exceeding 10 times the analytical cutoff for potential high-dose . Manual retesting protocols, often integrated into systems, recommend reflexive dilution for samples with borderline or unexpected results to mitigate false lows. The required dilution factor to resolve the hook effect correlates with its severity, with more extreme excess necessitating higher dilutions (e.g., 1:1000 or greater) for accurate quantitation. Validation of these identification methods follows Clinical and Laboratory Standards Institute (CLSI) guidelines, particularly EP07 for testing, which outlines protocols to screen, quantify, and confirm hook effects in clinical samples through systematic dilution and recovery assessments.

Prevention Methods

To prevent the hook effect in immunoassays, laboratories can adopt two-step protocols that separate the capture and detection phases. In these methods, the sample is first incubated with the capture to bind the , followed by a step to remove unbound excess before adding the labeled detection . This approach minimizes the from high concentrations by ensuring that free does not compete with the antibody-antigen complex during detection. Competitive immunoassay formats also inherently reduce the risk of the hook effect, particularly when incorporating a wash step after the initial analyte-capture antibody reaction. Unlike assays, competitive designs measure the unbound fraction of labeled , avoiding saturation issues at high analyte levels; this is programmed into many automated analyzers to extend the measurable for analytes prone to excess, such as tumor markers. Routine pre-dilution of samples suspected of high analyte concentrations serves as a proactive sample handling . For instance, implementing a standard 1:5 upfront dilution for ferritin assays has been shown to eliminate hook effect occurrences up to 126,050 µg/L, while also shortening turnaround times and reducing the need for testing. Similar 1:10 dilutions are recommended for tumor markers like or in clinical protocols to broaden the linear range without initial false lows. testing algorithms, where samples yielding results near the upper limit trigger automatic dilution, further integrate this into lab workflows for at-risk analytes. Technological advancements in automated systems enhance prevention through built-in design features. The platform employs a sequential format with an integrated washing step, significantly lowering the incidence of high-dose effects compared to one-step systems, with hook thresholds often exceeding clinical relevance for hormones like . Some modern analyzers incorporate software algorithms that monitor signal curvature for potential non-linearity, prompting preemptive adjustments, though primary prevention relies on architecture. Recent developments as of 2025 include the use of real-time reaction kinetics in lateral flow immunoassays to mitigate the effect by distinguishing true binding from saturation artifacts. Best practices emphasize rigorous lab validation aligned with manufacturer specifications, including hook effect testing during method verification to confirm the upper limit of linearity for each . Education for laboratory personnel on high-risk measurands, such as or , promotes vigilance in sample selection and protocol adherence. Professional organizations like the Clinical and Laboratory Standards Institute (CLSI) recommend evaluation, including high-dose testing, as part of routine method implementation to safeguard accuracy.

References

  1. [1]
    The Hook Effect | myadlm.org
    Oct 7, 2014 · The hook effect gives falsely low results in one-step immunoassays when excess analyte prevents the formation of the antigen-antibody sandwich.
  2. [2]
    Unraveling the Hook Effect: A Comprehensive Study of High Antigen ...
    Nov 13, 2020 · The hook effect results from the simultaneous reaction of an excess target antigens with both immobilized and labeled antibodies resp. To reduce ...Results and Discussion · Conclusions · Supporting Information · References
  3. [3]
    Hook Effect in Immunoglobulin A (IgA) Measurements: A Diagnostic ...
    Sep 16, 2025 · The high-dose hook effect, also known as the post-zone effect, occurs in immunoassays when extremely high concentrations of an analyte interfere ...
  4. [4]
    Mitigating the hook effect in lateral flow sandwich immunoassays ...
    The hook effect causes falsely low results in high concentrations. This paper uses reaction kinetics to monitor the test and control lines, increasing the ...
  5. [5]
    [PDF] Bioanalytical Method Validation - Guidance for Industry | FDA
    May 24, 2018 · Hook effect: The hook effect occurs when increasing analyte concentrations result in no change or decreased signals when compared to the ...
  6. [6]
    Retrospective Approach to Evaluate Interferences in Immunoassay
    The hook effect or the prozone effect is a type of interference which plagues certain immunoassays and nephelometric assays, resulting in false negatives or ...
  7. [7]
    Interferences in Immunoassay - PMC - PubMed Central - NIH
    Interfering substances may lead to falsely elevated or falsely low analyte concentration in one or more assay systems depending on the site of the interference ...
  8. [8]
    Immunoassay Methods - Assay Guidance Manual - NCBI Bookshelf
    May 1, 2012 · A sandwich immunoassay is a method using two antibodies, which bind to different sites on the antigen or ligand (Figure 1). The capture antibody ...
  9. [9]
    Immunoassay Methods and their Applications in Pharmaceutical ...
    The non-competitive design (usually called “two-site” or “sandwich” assay) is used for large analytes possessing more than one recognition epitopes on the ...
  10. [10]
    Interpretation of common endocrine laboratory tests - NIH
    Jul 24, 2019 · The shape of the binding curve gave the name “hook effect” to the phenomenon; with gradually-increasing analyte concentrations in the sample, ...
  11. [11]
    CDC Laboratory Recommendations for Syphilis Testing, United ...
    Feb 8, 2024 · This false-negative phenomenon is referred to as a prozone or hook effect because it occurs before the zone of equivalence where the ...
  12. [12]
    The Composition of the Precipitate in Precipitin Reactions - PMC
    Quantitative Aspects in Immunity Reactions: The Composition of the Precipitate in Precipitin Reactions · John Marrack · Frank Campbell Smith.
  13. [13]
    The Precipitation Reaction: Experiments with an Antiserum ... - jstor
    MARRACK, J. R. (1934). The c71,emistry of antigens and antibodies. M.R.C. Report, Chap. II. London: H.M. Stationery Office. MARRAax, J. and SMITH, F. C. (1931).
  14. [14]
    Mitigating hook effect in one-step quantitative sandwich lateral flow ...
    Apr 1, 2022 · First observed in one-step sandwich immunoassays in the 1980s, the hook effect is an intrinsic phenomenon in sandwich assays [9]. Assuming ...
  15. [15]
    Precipitation Reaction- Definition, Principle, Steps, Types, Uses
    May 21, 2022 · The zone below the zone of equivalence i.e. where antigens are lesser than antibodies is called the prozone phenomenon.Missing: 1900s | Show results with:1900s
  16. [16]
    Antibody-Mediated Immunomodulation: a Strategy To Improve Host ...
    Feb 1, 2005 · ... 10-fold antibody excess (29). Applications for infectious disease ... A prozone-like phenomenon has been described in passive ...
  17. [17]
    Interferences in quantitative immunochemical methods
    The hook effect - An excessive amount of analyte overwhelms the binding capacity of the capture antibody. This results in an inappropriately low signal that ...Abstract · Introduction · The Hook Effect
  18. [18]
    Hormone Immunoassay Interference: A 2021 Update - PMC
    This phenomenon may occur in sandwich immunoassays when measuring very high hormone concentrations; the hook effect occurs specifically in progressive tumor ...
  19. [19]
  20. [20]
    The hook effect: a need for constant vigilance - PubMed
    The hook effect is a well-recognized problem that can occur in assays of most tumour markers, including alpha feto-protein (AFP).Missing: PSA | Show results with:PSA
  21. [21]
    The Fundamental Flaws of Immunoassays and Potential Solutions ...
    Many assays have been reported to suffer from the hook effect in patients with high analyte values and include assays for prolactin (Fleseriu et al., 2006), ...
  22. [22]
    The Hook Effect: A Case Study of a Giant Invasive Prolactinoma With ...
    Sep 29, 2023 · The hook effect is a significant phenomenon to consider in the diagnosis and monitoring of giant and invasive prolactinomas.Missing: vivo | Show results with:vivo
  23. [23]
    Hook-effect in MAGLUMI immunoassay for serum anti-GAD ...
    May 15, 2024 · In this case report we found that anti-GAD testing in CSF was essential to establish the clinical diagnosis and to suspect hook-effect in serum ...
  24. [24]
    High-Dose Hook Effect in Patients with Macroprolactinoma.
    High-Dose Hook Effect in Patients with Macroprolactinoma. ... Our retrospective study also involved 13 patients with macroprolactinoma for the comparison of the ...Missing: vivo | Show results with:vivo
  25. [25]
    Ovarian choriocarcinoma: a difficult diagnosis of an unusual tumor ...
    Because of falsely low beta-hCG values (283 mIU/L) caused by the high-dose "hook effect" in the double antibody immunoassay system, however, a diagnosis of ...
  26. [26]
    Gestational Trophoblastic Disease Diagnosis Delayed by the Hook ...
    A "hook effect" resulting from saturation of antibodies used in pregnancy tests can occur at human chorionic gonadotropin (hCG) levels above 500,000 ...Missing: beta- 2000<|separator|>
  27. [27]
    Hook effect in gestational trophoblastic disease: An emergency ...
    Oct 25, 2022 · The authors discuss the presentation and diagnostic process of a patient in the emergency department as well as the phenomenon known as the hook effect.Missing: 2000 | Show results with:2000
  28. [28]
    Hook Effect - an overview | ScienceDirect Topics
    The Prozone or hook effect is observed when a very high amount of an analyte is present in the sample but the observed value is falsely lowered. This type of ...Missing: origin | Show results with:origin
  29. [29]
    High-Dose Hook Effect in Patients with Macroprolactinoma
    The "hook effect": high dose concentrations of prostatespecific antigen giving artifactually low values on one-step immunoassay. N Engl J Med. 1989. 320 ...
  30. [30]
    “Hook Effect” in Prolactinomas: Case Report and Review of Literature
    N Engl J Med. 1989; 320:1755-1756. Crossref · Scopus (21) · PubMed · Google Scholar. 37. Myrtle, J.F.. More on “hook effects” in immunometric assays for ...
  31. [31]
    Final Diagnosis -- High-dose hook effect or prozone effect
    High-dose Hook effect. The hook effect is a common interference encountered in immunoassay techniques.
  32. [32]
    A Practical Guide to Immunoassay Method Validation - Frontiers
    Thereby dilution of samples should not affect the accuracy and precision. At the same time, the presence of a hook effect, i.e., suppression of signal at ...
  33. [33]
    [PDF] Elecsys Troponin T hs | Roche
    There is no high-dose hook effect at troponin T concentrations up to. 100000 ng/L (pg/mL). Pharmaceutical substances. In vitro tests were performed on 17 ...
  34. [34]
    Resolution of an Apparent Hook Effect in Roche Partner DRI ...
    May 27, 2013 · A presumed hook effect in the semiquantitative DRI Oxycodone immunoassay, OXY3S (Cobas Integra, Roche Diagnostics), was investigated in 14 urine ...
  35. [35]
    Upfront dilution of ferritin samples to reduce hook effect ... - PubMed
    Feb 15, 2018 · Conclusions: Implementation of upfront dilutions of all ferritin samples reduced the possibility of hook effect, improved turnaround time and ...
  36. [36]
    High-dose hook effect in six automated human chorionic ...
    Miles et al. first described high-dose hook effect in the literature in the early 1970s with the two-site immunoradiometric assay for ferritin,3 and it remains ...
  37. [37]
    Guidelines on Clinical Method Validation & Verfication - CLSI
    Feb 18, 2019 · CLSI offers an entire library of method evaluation standards and method verification guidelines that provide concise explanations and step-by-step instructions.