Detection of Fish Antigens Aerosolized during Fish Processing Using Newly Developed Immunoassays

Abstract

Background: Aerosolization of fish proteins during seafood processing has been identified as a potential route for allergic sensitization and occupational asthma among workers involved in high-risk activities. The aim of this study was to develop immunological assays for the quantification of aerosolized fish antigens in a fish-processing factory. Methods: Polyclonal antibodies to the main fish species processed in the factory (anchovy and pilchard) were generated in rabbits and compared by ELISA inhibition assay and immunoblotting. These antisera were utilized to develop ELISA assays for the detection of fish antigens. The ELISA inhibition assays were evaluated by analyzing environmental air samples collected from three areas in a fish-processing factory: pilchard canning, fish meal production and lobster processing. Results: By immunoblotting, the rabbit polyclonal antibodies demonstrated IgG antibody binding patterns comparable with IgE antibodies of fish-sensitized patients, particularly in regard to the major fish allergens parvalbumins. The sensitivity of the fish-specific ELISA assays developed was 0.5 µg/ml. The ELISA inhibition assays were able to differentiate between the two different fish species of interest but did not recognize a crustacean species. Notable differences in exposure levels to canned pilchard and anchovy antigens were demonstrated in the three different working areas of the factory, with assays having a detection limit as low as 105 ng/m³. Conclusion: These ELISA-based assays are sensitive and specific to quantify differential exposure levels to fish antigens produced during fish processing, making it possible to investigate exposure-disease response relationships among workers in this industry.

Important Variations in Parvalbumin Content in Common Fish Species: A Factor Possibly Contributing to Variable Allergenicity

“…This allowed to detect parvalbumins with high sensitivity and accuracy in the samples. Previously, it has been shown that the use of non-species-specific antibodies carries pitfalls [12,14,15]. In this study, the calibration curves clearly demonstrate a difference in antibody sensitivity (fig.…”

Occupational Exposure to Bioaerosols in Norwegian Crab Processing Plants

“…The sensitive ELISA method used to measure tropomyosin (Kamath et al 2014;Malo et al 1997) has no unspecific binding to other proteins and no loss of binding due to protein modification procedures that may have influenced the levels reported in other studies (Griffin P 1994;Lopata and Jeebhay 2013;Lopata et al 2005). Previous studies have also used the highly sensitive mass spectrometric based strategies to quantify tropomyosin and arginine kinase (Rosmilah et al 2012;Abdel Rahman, Gagne, and Helleur 2012;Abdel Rahman et al 2011;Abdel Rahman et al 2010).…”

Fish Allergen Detection

“…The antigenicity of a protein is not necessarily like its allergenicity; however, IgG-antibodies are useful to develop immunoassays for the detection of food allergens. Polyclonal antibodies against canned pilchard and fresh anchovy were produced to develop an inhibition enzyme-linked immunosorbent assay (ELISA) for the detection of aerosolized fish antigens (Lopata et al, 2005). The assay sensitivity was 0.5 mg/mL in phosphate-buffered saline/0.05% Tween-20 extracts of environmental air samples correlating to a 105-ng/m 3 aerosol concentration.…”

“…However, if fish allergen is inhaled (e.g., in the vapor from boiling fish or in the air at a fish market), symptoms of asthma and even generalized anaphylactic symptoms may develop (Crespo et al, 1995;. In fish-processing workplaces, occupational exposure to fish allergens can induce the typical IgE-mediated hypersensitive systemic, cutaneous, gastrointestinal, and respiratory reactions (Douglas et al, 1995;Rodr ıguez et al, 1997;Sicherer et al, 2000;Lopata et al, 2005).…”