Molecular and immunological approaches in quantifying the air-borne food allergen tropomyosin in crab processing facilities

Kamath, Sandip D., Thomassen, Marte R., Saptarshi, Shruti R., Nguyen, Hong M.X., Aasmoe, Lisbeth, Bang, Berit E., and Lopata, Andreas L. (2014) Molecular and immunological approaches in quantifying the air-borne food allergen tropomyosin in crab processing facilities. International Journal of Hygiene and Environmental Health, 217 (7). pp. 740-750.

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Abstract

Tropomyosin is a cross-reactive allergenic protein present in ingested shellfish species. Exposure and sensitization to this protein via inhalation is particularly important in the crustacean processing industry where workers are continuously exposed to the aerosolized form of this allergen. The aim of this study was to develop an antibody-based immunoassay to enable the specific and sensitive quantification of aerosolized tropomyosin present in the environment of two crab processing facilities. Anti-tropomyosin antibody was generated in rabbits against tropomyosins from four different crustacean species. These antibodies were purified using recombinant tropomyosin using an immuno-affinity column. The recombinant tropomyosin was also used as an allergen standard for the sandwich ELISA. In order to quantify aerosolized tropomyosin, air collection was performed in the personal breathing zone of 80 workers during two crab processing activities, edible crab (Cancer pagurus) and king crab (Paralithodes camtschaticus) using polytetrafluoroethylene filters. The purified antibody was able to detect tropomyosin selectively from different crustaceans but not from vertebrate sources. The limit of detection (LOD) for the developed sandwich ELISA was 60picogram/m(3) and limit of quantitation (LOQ) 100picogram/m(3). Immunoassay validation was based on linearity (R(2) 0.999), matrix interference test (78.8±6.5%), intra-assay CV (9.8%) and inter-assay CV (11%). The novel immunoassay was able to successfully identify working activities, which generated low, medium or high concentrations of the aerosolized food allergen. We describe an IgG antibody-based immunoassay for quantification of the major food allergen tropomyosin, with high sensitivity and specificity. This modified immunological approach can be adapted for the detection of other aerosolized food allergens, assisting in the identification of high-risk allergen exposure areas in the food industry.

Item ID: 34331
Item Type: Article (Research - C1)
ISSN: 1618-131X
Keywords: food allergy; IgE antibody; allergen; seafood; immunology; occupational allergy
Funders: Norwegian Asthma and Allergy Association, Australian Research Council (ARC)
Research Data: http://dx.doi.org/10.4225/28/588e9d7db86da
Date Deposited: 07 Aug 2014 04:23
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110701 Allergy @ 50%
11 MEDICAL AND HEALTH SCIENCES > 1117 Public Health and Health Services > 111705 Environmental and Occupational Health and Safety @ 50%
SEO Codes: 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920108 Immune System and Allergy @ 100%
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