title: Probing the interaction of benzo[a]pyrene adducts and metabolites with monoclonal antibodies using fluorescence line-narrowing spectroscopy.
abstract: A new approach for studying antibody-antigen interactions of DNA adducts and metabolites of polycyclic aromatic hydrocarbons (PAHs) is demonstrated in which fluorescence line-narrowing spectroscopy (FLNS) is used. It is based on the fact that in an FLN spectrum the relative intensities of the line-narrowed bands (that correspond to the excited-state vibrations) are, in general, strongly dependent on the local environment of the fluorophore. Information on the nature of the interactions can be obtained by comparing the FLN spectra of the antigen-antibody complexes to the spectra of the antigen in different types of solvents (H-bonding, aprotic, and pi-electron-containing solvent molecules) recorded under the same conditions. The antigens used were the DNA adduct 7-(benzo[a]pyren-6-yl)guanine (BP-6-N7Gua) and the metabolite (+)-trans-anti-7,8,9,10-benzo[a]pyrenetetrol (BP-tetrol) of benzo[a]pyrene; two monoclonal antibodies (MAbs) have been developed to selectively bind these compounds. It is shown that, for BP-tetrol, H-bonding solvents have a pronounced effect on the FLN spectra. The presence of pi electrons in the solvent molecules results in relatively small but still significant changes in the spectra. When BP-tetrol is bound to its MAb, however, neither of these effects is observed; its spectrum is very similar to the one obtained with an aprotic solvent, methylcyclohexane. Therefore, we can conclude that this MAb has an internal binding site in which the interaction with BP-tetrol is of a hydrophobic character. For BP-6-N7Gua, however, there is a strong effect of the presence of pi electrons in the solvent molecules. The FLN spectrum of this antigen bound to its MAb is very similar to its spectrum in acetone, indicating that pi-pi interactions play an important role in the binding.
authors.affiliation: Department of Analytical Chemistry and Applied Spectroscopy, Laser Centre, Vrije Universiteit, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: Arjen N
authors.initials: AN
authors.lastname: Bader
authors.forename: Nenad M
authors.initials: NM
authors.lastname: Grubor
authors.forename: Freek
authors.initials: F
authors.lastname: Ariese
authors.forename: Cees
authors.initials: C
authors.lastname: Gooijer
authors.forename: Ryszard
authors.initials: R
authors.lastname: Jankowiak
authors.forename: Gerald J
authors.initials: GJ
authors.lastname: Small
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D001564:Benzo(a)pyrene; D001580:Benzopyrenes; D018736:DNA Adducts / Q000276:immunology*; D006147:Guanine / Q000031:analogs & derivatives* / Q000276:immunology; D006860:Hydrogen Bonding; D015394:Molecular Structure; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: C102824:7-(benzo(a)pyren-6-yl)guanine; D000911:Antibodies, Monoclonal; D001580:Benzopyrenes; D018736:DNA Adducts; C056612:benzo(a)pyrene-DNA adduct; D001564:Benzo(a)pyrene; D006147:Guanine; C040059:7,8,9,10-tetrahydrobenzo(a)pyrene