The interaction of highly cross-reactive monoclonal antibodies (mAb) with four diastereomeric benzo[a]pyrene tetrols (BPTs) and benzo[a]pyrene diolepoxide (BPDE) derived deoxyguanosine (dG) adducts is studied by means of fluorescence line-narrowing spectroscopy (FLNS). It is shown that the interactions of different enantiomers (i.e. (+/-)-cis and (+/-)-trans) of these compounds with promiscuous monoclonal antibodies involve different complex geometries. These spatially different ligand-protein interactions alter the relative intensities of the excited-state vibrational frequencies of immunocomplexed molecules, allowing for unambiguous spectroscopic resolution of all four enantiomeric isomers. In addition, immunoaffinity capillary electrophoretic (IACE) separation of all four stereisomers of BPT and of BPDE-dG DNA adducts has been achieved using polycyclic anti-aromatic hydrocarbon (anti-PAH) mAb and 8E11 mAb, respectively. This study confirms that a high-resolution FLNS approach is capable of enantiospecific differentiation and could be used to discriminate among various enantiomers immunocomplexed with various monoclonal antibodies.