Analytical and Bioanalytical Chemistry

, Volume 409, Issue 25, pp 5999–6014 | Cite as

Cross-reactivity profiles of legumes and tree nuts using the xMAP® multiplex food allergen detection assay

  • Chung Y. Cho
  • Carolyn Oles
  • William Nowatzke
  • Kerry Oliver
  • Eric A.E. GarberEmail author
Research Paper


The homology between proteins in legumes and tree nuts makes it common for individuals with food allergies to be allergic to multiple legumes and tree nuts. This propensity for allergenic and antigenic cross-reactivity means that commonly employed commercial immunodiagnostic assays (e.g., dipsticks) for the detection of food allergens may not always accurately detect, identify, and quantitate legumes and tree nuts unless additional orthogonal analytical methods or secondary measures of analysis are employed. The xMAP® Multiplex Food Allergen Detection Assay (FADA) was used to determine the cross-reactivity patterns and the utility of multi-antibody antigenic profiling to distinguish between legumes and tree nuts. Pure legumes and tree nuts extracted using buffered detergent displayed a high level of cross-reactivity that decreased upon dilution or by using a buffer (UD buffer) designed to increase the stringency of binding conditions and reduce the occurrence of false positives due to plant-derived lectins. Testing for unexpected food allergens or the screening for multiple food allergens often involves not knowing the identity of the allergen present, its concentration, or the degree of modification during processing. As such, the analytical response measured may represent multiple antigens of varying antigenicity (cross-reactivity). This problem of multiple potential analytes is usually unresolved and the focus becomes the primary analyte, the antigen the antibody was raised against, or quantitative interpretation of the content of the analytical sample problematic. The alternative solution offered here to this problem is the use of an antigenic profile as generated by the xMAP FADA using multiple antibodies (bead sets). By comparing the antigenic profile to standards, the allergen may be identified along with an estimate of the concentration present. Cluster analysis of the xMAP FADA data was also performed and agreed with the known phylogeny of the legumes and tree nuts being analyzed.

Graphical abstract

The use of cluster analysis to compare the multi-antigen profiles of food allergens


Food allergens Legumes Tree nuts xMAP multiplex assay 



Our gratitude is expressed to Masahiro Shoji, Ph.D. (Morinaga Institute of Biological Sciences, Inc.); Mansour Samadpour, Ph.D. (IEH Laboratories and Consulting Group); and Thomas Grace (Elution Technologies) for their openness and willingness to make resources available for the xMAP FADA. Gratitude is also expressed to Shaun MacMahon, Ph.D.; George C. Ziobro, Ph.D.; Prasad Rallabhandi, Ph.D.; Ronnie O. Pedersen, Ph.D. (FDA); and Lynn L.B. Rust (NIH) for the valuable discussions.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2017_528_MOESM1_ESM.pdf (143 kb)
ESM 1 (PDF 143 kb)


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Copyright information

© US Government (outside the USA) 2017

Authors and Affiliations

  • Chung Y. Cho
    • 1
  • Carolyn Oles
    • 1
  • William Nowatzke
    • 2
  • Kerry Oliver
    • 2
  • Eric A.E. Garber
    • 1
    Email author
  1. 1.Office of Regulatory Science, Center for Food Safety and Applied Nutrition (CFSAN)Food and Drug AdministrationCollege ParkUSA
  2. 2.Radix® BioSolutionsGeorgetownUSA

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