Health  Vol.8 No.15 , December 2016
A Simple Method for Detection of Multiple Chemical-Specific IgGs in Serum Based on Dot Blotting
Abstract: Plastic resins are known to cause occupational allergies. Therefore, serum-specific antibodies against plastic resins have been widely investigated as diagnostic markers for occupational allergies. In this study, we aimed to establish a convenient method for detection of multiple chemical-specific IgG antibodies in human serum based on dot blot analysis. Toluene diisocyanate (TDI), phthalic anhydride (PA), and formaldehyde (FA), which are frequently used to synthesize various resins, reacted well with lysine residues of human serum albumin (HSA) under alkaline conditions. Native polyacrylamide gel electrophoresis (PAGE) showed that the structures of chemical adducts of HSA were different from those of native HSA. Therefore, we performed dot blot assays using these adducts as artificial antigens. Serum samples from workers at plants utilizing plastic resins strongly reacted with TDI, PA, and FA adducts in HSA, while reduced signals were detecting using the serum from unexposed workers. These results suggested that dot blot assays using chemical-HSA adducts as antigens could be beneficial for simultaneously measuring multiple chemical-specific IgGs.
Cite this paper: Tsuji, M. , Yu, H. , Ishihara, Y. , Isse, T. , Ikeda-Ishihara, N. , Tuchiya, T. and Kawamoto, T. (2016) A Simple Method for Detection of Multiple Chemical-Specific IgGs in Serum Based on Dot Blotting. Health, 8, 1645-1653. doi: 10.4236/health.2016.815160.

[1]   Plastics Europe (2011) Plastics—The Facts 2011. An Analysis of European Plastics Production, Demand and Recovery for 2010.

[2]   Lithner, D., Larsson, A. and Dave, G. (2011) Environmental and Health Hazard Ranking and Assessment of Plastic Polymers Based on Chemical Composition. The Science of the Total Environment, 409, 3309-3324.

[3]   Palikhe, N.S., Kim, J.H. and Park, H.S. (2011) Biomarkers Predicting Isocyanate-Induced Asthma. Allergy, Asthma & Immunology Research, 3, 21-26.

[4]   de Groot, A.C., White, I.R., Flyvholm, M.A., Lensen, G. and Coenraads, P.J. (2010) Formaldehyde-Releasers in Cosmetics: Relationship to Formaldehyde Contact Allergy. Part 1. Characterization, Frequency and Relevance of Sensitization, and Frequency of Use in Cosmetics. Contact Dermatitis, 62, 2-17.

[5]   Kadivar, S. and Belsito, D.V. (2015) Occupational Dermatitis in Health Care Workers Evaluated for Suspected Allergic Contact Dermatitis. Dermatitis: Contact, Atopic, Occupational. Drug, 26, 177-183.

[6]   Bernstein, D.I., Ott, M.G., Woolhiser, M., Lummus, Z. and Graham, C. (2006) Evaluation of antibody Binding to Diisocyanate Protein Conjugates in a General Population. Annals of Allergy, Asthma & Immunology: Official Publication of the American College of Allergy, Asthma, & Immunology, 97, 357-364.

[7]   Sashidhar, R.B., Capoor, A.K. and Ramana, D. (1994) Quantitation of Epsilon-Amino Group Using Amino Acids as Reference Standards by Trinitrobenzene Sulfonic Acid. A Simple Spectrophotometric Method for the Estimation of Hapten to Carrier Protein Ratio. Journal of Immunological Methods, 167, 121-127.

[8]   Pham, T.T., Oyama, T., Isse, T. and Kawamoto, T. (2009) Application of Tryptophan Fluorescence to Assess Sensitizing Potentials of Chemicals. Archives of Environmental Contamination and Toxicology, 57, 427-436.

[9]   Kawamoto, T., Tsuji, M. and Isse, T. (2015) Comparison of IgG against Plastic Resin in Workers with and without Chemical Dermatitis. BMC Public Health, 15, 930.

[10]   Ott, M.G., Jolly, A.T., Burkert, A.L. and Brown, W.E. (2007) Issues in Diisocyanate Antibody Testing. Critical Reviews in Toxicology, 37, 567-585.

[11]   Ye, Y.M., Kim, C.W., Kim, H.R., Kim, H.M., Suh, C.H., Nahm, D.H., et al. (2006) Biophysical Determinants of Toluene Diisocyanate Antigenicity Associated with Exposure and Asthma. The Journal of Allergy and Clinical Immunology, 118, 885-891.

[12]   Rosqvist, S., Nielsen, J., Welinder, H., Rylander, L., Lindh, C.H. and Jonsson, B.A. (2003) Exposure-Response Relationships for Hexahydrophthalic and Methylhexahydrophthalic Anhydrides with Total Plasma Protein Adducts as Biomarkers. Scandinavian Journal of Work, Environment & Health, 29, 297-303.

[13]   Yarbrough, J.W. and Schultz, T.W. (2007) Abiotic Sulfhydryl Reactivity: A Predictor of Aquatic Toxicity for Carbonyl-Containing Alpha, Beta-Unsaturated Compounds. Chemical Research in Toxicology, 20, 558-562.