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| Article Authors: | Tinashe B Ruwona; Victor J Johnson; Detlef Schmechel; Reuben H Simoyi; Donald Beezhold; Paul D Siegel |
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| Article Title: | Monoclonal antibodies against toluene diisocyanate haptenated proteins from vapor-exposed mice. |
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| Reference ID: | 1019883 |
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| Abstract: | Toluene diisocyanate (TDI) is an industrially important polymer cross-linker used in the production of polyurethane. Workplace exposure to TDI and other diisocyanates is reported to be a leading cause of low molecular weight-induced occupational asthma (OA). Currently we have a limited understanding of the pathogenesis of OA. Monoclonal antibodies (MAbs) that recognize TDI bound proteins would be valuable tools/reagents, both in exposure monitoring and in TDI-induced asthma research. We sought to develop toluene diisocyanate (TDI)-specific MAbs for potential use in the development of standardized immunoassays for exposure and biomarker assessments. Mice were exposed 4 h/day for 12 consecutive weekdays to 50 ppb, 2,4;2,6 TDI vapor (80/20 mixture). Splenocytes were isolated 24 h after the last exposure for hybridoma production. Hybridomas were screened in a solid-phase indirect enzyme-linked immunosorbent assay (ELISA) against a 2,4 TDI-human serum albumin (2,4 TDI-HSA) protein conjugate. Three hybridomas producing 2,4 TDI-HSA reactive IgM MAbs were obtained. The properties of these MAbs (isotype and reactivity to various protein-isocyanate conjugate epitopes) were characterized using ELISA, dot blot, and Western blot analyses. Western blot analyses demonstrated that some TDI conjugates form inter- and intra-molecular links, resulting in multimers and a change in the electrophoretic mobility of the conjugate. These antibodies may be useful tools for the isolation of endogenous diisocyanate-modified proteins after natural or experimental exposures and for characterization of the toxicity of specific dNCOs. |
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| Affiliations: | Allergy and Clinical Immunology, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505-2888, USA. |
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| Date: | 2010 |
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| Reference Type: | Literature |
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| PubMed ID: | 20568997 |
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| Journal: | Hybridoma (Larchmt) |
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| Journal Volume: | 29 |
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| Article Pages: | 221-9 |
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| Journal ISSN: | 1554-0014 |
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| Article Chemical List: | Antibodies, Monoclonal;Haptens;Immunoglobulin M;Isocyanates;Serum Albumin;Toluene 2,4-Diisocyanate |
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| Article MeSH List: | Administration, Inhalation; Animals; Antibodies, Monoclonal(classification; diagnostic use; immunology); Antibody Specificity; Asthma(chemically induced; diagnosis); Enzyme-Linked Immunosorbent Assay; Female; Haptens(immunology); Humans; Hybridomas; Immunoblotting; Immunoglobulin M; Isocyanates(immunology); Mice; Mice, Inbred C57BL; Serum Albumin; Toluene 2,4-Diisocyanate(administration & dosage; immunology; toxicity) |
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| Curation Last Updated: | 2013-05-28 22:14:27 |
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