title: Characterization of antibodies against benzo[a]pyrene with thermodynamic and kinetic constants.
abstract: Antibodies of a polyclonal antiserum against benzo[a]pyrene were characterized by determining thermodynamic and kinetic constants of the antigen-antibody reaction. Label-free binding assays with optical detection based on reflectometric interference spectroscopy were performed to determine these constants. Different evaluation methods for kinetic measurements were compared. Also, cross-reactivity against two other polycyclic aromatic hydrocarbons, chrysene and pyrene, was checked. The affinity constant between the antibodies and benzo[a]pyrene in homogeneous phase was determined to be K=(5.3+/-0.3)x10(7) M(-1) which was in the middle of the usual range of antibody affinities. The association rate constant for the reaction at the surface was determined to be (3.8+/-0.9)x10(5) M(-1) s(-1), the dissociation rate constant as (9.7+/-0.5)x10(-3) s(-1). Different evaluation methods applied to the kinetic measurements led to the same results. This antiserum would be suitable for the selective determination of benzo[a]pyrene in concentrated samples.
authors.affiliation: Institute of Physical and Theoretical Chemistry, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Länge
authors.forename: G
authors.initials: G
authors.lastname: Griffin
authors.forename: T
authors.initials: T
authors.lastname: Vo-Dinh
authors.forename: G
authors.initials: G
authors.lastname: Gauglitz