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

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FRNKOVÁ Kateřina PŘIBYL Jan SKLÁDAL Petr SUCHÁNEK Miloslav

Rok publikování 2008
Druh Článek ve sborníku
Konference 5th International Interdisciplinary Meeting on Bioanalysis CECE2008
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Obor Biochemie
Klíčová slova bioconjugates immunization ELISA
Popis 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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