Toluene diisocyanate (TDI) is a commonly used diisocyanate (dNCO). TDI is a reactive low molecular weight chemical that is widely used in industry, especially in the manufacture of polyurethane foams and adhesives. Diisocyanate exposure is one of the most commonly reported causes of occupational asthma. The production of well characterized TDI-specific monoclonal antibodies (mAbs) will allow for the development of standardized immunoassays for exposure and biomarker assessment. Such mAbs may also be useful to isolate and study dNCO protein targets. BALB/c mice were immunized with 2,4- or 2,6-TDI conjugated keyhole limpet hemocyanin (KLH), spleen cells isolated and hybridomas made. Resultant mAbs produced by these hybridomas were screened for their ability to bind TDI conjugated human serum albumin (HSA) by ELISA. MAbs were further characterized by ELISA and western blot against various monoisocyanate (mNCO) and dNCO and diisothiocyanate conjugated proteins to identify reactivity toward urea, amide or thiourea linkages. A total of 35 mAbs were produced (25 IgG1, 8 IgG2a, 2 IgG2b) were obtained against 2,4-TDI-KLH sensitized mice. Seven mAbs were found to recognize 2,4-TDI-HSA and 28 mAbs reacted with both 2,4- and 2,6- TDI-HSA. mAbs specific for only 2,4-TDI-conjugated protein did not recognize 2,6-TDI-HSA, mNCO-HSA or other dNCO-HSAs. Other cross-reactive mAbs reacted with 2,4-TDI-HSA, 2,6-TDI-HSA, and other dNCO protein conjugates like methylene diphenyldiisocyanate- and hexamethelene diisocyanate-HSA, but not mNCO-HSA. All mAb reactivities were carrier protein independent. The mAbs produced can differentiate between specific dNCO in a carrier independent fashion and may be useful as hapten-specific reagents for immunoassay development and research into dNCO related diseases.
Biological-factors; Biomarkers; Breathing; Chemical-hypersensitivity; Employee-exposure; Exposure-assessment; Exposure-levels; Exposure-methods; Lung-irritants; Occupational-exposure; Occupational-hazards; Occupational-health; Pulmonary-system-disorders; Inhalation-studies; Respiratory-hypersensitivity; Respiratory-system-disorders; Risk-factors; Risk-analysis; Toxic-effects
The Toxicologist. Society of Toxicology 48th Annual Meeting and ToxExpo, March 15-19, 2009, Baltimore, Maryland