Tinashe Ruwona
Present Work and Research

The   goal of my research is to develop monoclonal antibodies specific for toluene diisocyanate conjugated proteins. These monoclonal antibodies can be used for diagnosis, immunohistochemistry and biomarker immunoassays of diisocyanates exposure.  

Background: 

Diisocyanates (dNCOs) are used in the production of polyurethanes. The most common monomeric dNCOs are toluene diisocyanate (TDI), methylene diphenyl diisocyanate (MDI) and hexamethylene diisocyanate (HDI).  Workers are exposed to these, as well as polymeric forms of diisocyanates.  They are also the most commonly reported cause of occupational asthma.  They can bind to proteins through a number of chemical functional groups and the nature of this binding may depend on the protein and the local environment.  It has been shown that dNCOs readily complex with the proteins; human serum albumin (HSA) and hemoglobin.  In murine studies, dNCO-bound protein has been found co-localized in the lung with tubulin, actin and keratin by immunochemical staining which suggest that these proteins may also be conjugate.

The objective of my work is to produce monoclonal IgGs that will recognize 2,4- and 2,6-TDI-bound proteins. These monoclonal antibodies can be used for diagnosis, immunohistochemistry and for biomarker immunoassays of dNCO exposure.  In addition, they should make useful research tools in identifying specific dNCO-bound proteins following both dermal and inhalation exposures.

In our initial work we have taken spleen and lymph nodes from TDI-vapor exposed mice and obtained several monoclonal antibodies that recognize TDI bound mouse albumin.  All the antibodies that we obtained were of the class, IgM, which have limited usefulness due their propensity to precipitate out of solution.  The most useful class of monoclonals is IgG which are highly soluble, stable and have high specificity.

 We hypothesize that we should be able to produce monoclonal IgGs that will recognize general TDI bound proteins. It may be possible, also, to produce antibodies that are specific to the type of binding (i.e. binding of dNCO to an amine to form a urea vs. to a hydroxyl group to form an ester).

This work is done at the CDC/NIOSH in Morgantown in collaboration with  Capt P.D Siegel PhD.

This work is supported by NIEHS and NSF.

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