Cross-reactivity in antibody microarrays and multiplexed sandwich assays: shedding light on the dark side of multiplexing

Juncker, David; Bergeron, Sébastien; Laforte, Véronique; Li, Huiyan

doi:10.1016/j.cbpa.2013.11.012

Cited by 118 publications

(116 citation statements)

References 43 publications

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“…In the commercial systems, the preference is given to mono clonal antibodies because their properties are more reproducible. Recently, studies on the modification of the properties of natural antibodies (for example, the use of recombinant single strand variable fragments of antibodies or chimeric antibodies, in which a portion of proteins is replaced by human protein components) and, as an alternative to them, the development of so called artificial antibodies, for example, aptamers and polymers with molecular imprints have been per formed [28].…”

Section: Immunochemical Methods Usedmentioning

confidence: 99%

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Contemporary trends in the development of immunochemical methods for medical analysis

Goryacheva

2015

J Anal Chem

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The review is dedicated to contemporary trends in the development of immunochemical meth ods, the position of these methods in clinical analysis, and the applications of these methods. The tendencies toward miniaturization and the simultaneous determination of several analytes are considered. The prospects of applying nanosized markers, the requirements imposed on them, and limitations are discussed in detail. Predictions of the subsequent development of immunochemical methods in the area of clinical analysis are made.

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Section: Immunochemical Methods Usedmentioning

confidence: 99%

“…Multicomponent analysis is especially popular in the context of the development of omics (genomics, proteomics, and metabolomics) [28]. From the standpoint of clinical analysis, the most important advantages of the simultaneous determina tion of several analytes consist in the following [1]:…”

Section: Multicomponent Analysismentioning

confidence: 99%

Contemporary trends in the development of immunochemical methods for medical analysis

Goryacheva

2015

J Anal Chem

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“…Multiplex immunoassays are particularly susceptible to false positives arising from cross-reactivity [59]. Cross-reaction can occur in several different ways: antibody-antibody, antibody-antigen and antigenantigen.…”

Section: Assay Characterizationmentioning

confidence: 99%

Systems for Multiplexing Homogeneous Immunoassays

2015

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High-throughput multiplex protein biomarker assays continue to gain significance in the fields of biomarker discovery and drug development, due to their economical use of not only the precious clinical biological samples but also expensive reagents. Among these platforms, homogeneous multiplex systems have potential for short assay run times and cost-effective reagent consumptions. However, these systems must overcome challenges of signal cross talk and biochemical cross-reactivity. Despite these obstacles, several homogeneous multiplex immunoassays have been demonstrated. These include fluorescent polarization, fluorescent resonance energy transfer with quantum dots or graphene, luminescent oxygen-channeling immunoassay coupled with aqueous two-phase systems and DNA proximity assays. The balance between speed/simplicity and high multiplexing and robustness of these homogeneous multiplex immunoassays are discussed in this review.

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“…This issue becomes more problematic as the number of antibody pairs in the assay increases, as the likelihood of nonspecific interactions will increase exponentially 7 . To mitigate risks of cross-reactions, tedious systematic evaluations of nonspecific interactions between each antibody and each measured protein must be performed to validate these assays 7–10 . Alternatively, matched capture and detection antibody pairs can also be co-localized 11,12 .…”

Section: Introductionmentioning

confidence: 99%

Aqueous two-phase systems enable multiplexing of homogeneous immunoassays

et al. 2014

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Quantitative measurement of protein biomarkers is critical for biomarker validation and early disease detection. Current multiplex immunoassays are time consuming costly and can suffer from low accuracy. For example, multiplex ELISAs require multiple, tedious, washing and blocking steps. Moreover, they suffer from nonspecific antibody cross-reactions, leading to high background and false-positive signals. Here, we show that co-localizing antibody-bead pairs in an aqueous two-phase system (ATPS) enables multiplexing of sensitive, no-wash, homogeneous assays, while preventing nonspecific antibody cross-reactions. Our cross-reaction-free, multiplex assay can simultaneously detect picomolar concentrations of four protein biomarkers ((C-X-C motif) ligand 10 (CXCL10), CXCL9, interleukin (IL)-8 and IL-6) in cell supernatants using a single assay well. The potential clinical utility of the assay is demonstrated by detecting diagnostic biomarkers (CXCL10 and CXCL9) in plasma from 88 patients at the onset of the clinical symptoms of chronic graft-versus-host disease (GVHD).

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Cross-reactivity in antibody microarrays and multiplexed sandwich assays: shedding light on the dark side of multiplexing

Cited by 118 publications

References 43 publications

Contemporary trends in the development of immunochemical methods for medical analysis

Contemporary trends in the development of immunochemical methods for medical analysis

Systems for Multiplexing Homogeneous Immunoassays

Aqueous two-phase systems enable multiplexing of homogeneous immunoassays

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