Publication details

Preparation of protein conjugates with highly hydrophobic molecules and their use in heterogeneous immunoassay



Year of publication 2008
Type Article in Proceedings
Conference 5th International Interdisciplinary Meeting on Bioanalysis CECE2008
MU Faculty or unit

Faculty of Science

Field Biochemistry
Keywords bioconjugates immunization ELISA
Description Chromatogpraphic methods coupled to a mass spectroscopy are widely used in analysi of PAHs and their metabolites, as those methods provide high sensitivity and selectivity, combined with excellent robustness. Immunoanalytical methods, such as ELISA, offer interesting possibility to detect PAHs at low levels with low expanses. Moreover, development of an antibody with specifity to polyaromatic hydrocarbons offers a possibility to detect DNA-PAH adducts. Preparation and characterization of immunoconjugates of polyaromatic hydrocarbon model compound (pyrenebutyric acid) with proteins (carrier proteins and enzymes) is described (shown as scheme on this page). PAHs compounds as highly lipophilic molecules can not be conjugated to a protein structure in a single step procedure, however, step-by-step changing of conjugation and purification steps needs to be employed in this way. Use of prepared conjugates in construction of direct and indirect ELISA method is shown, too.
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