Toluene diisocyanate vapor at 50 ± 5 ppb (TDI, Mondur TD80 Grade A, 80% and 20% mixture of 2,4- and 2,6-TDI isomers, respectively) provided by Bayer Corporation (Polyurethanes Division, Pittsburgh, PA).
Method of Immunization
Mice in a stainless steel live-in chamber (Unifab Corp., Kalamazoo, MI) were exposed to 50 ± 5 ppb TDI vapor for 4 h/day, for 12 consecutive business days.(1)
Parental Cell Line Used for Fusion
Murine Sp2/0 myeloma cells and splenocytes/lymph nodes from TDI vapor exposed mice.(2)
Selection and Cloning Procedure
Positive hybrids were selected by ELISA and then subcloned twice using limiting dilution.
Heavy and Light Chains of Immunoglobulin
IgM
Specificity
The specificity of these antibodies to detect the resultant epitopes formed from covalent binding of diisocyanates to protein nucleophilic groups was confirmed by ELISA and Western blot analyses.
