Κυανουρικό οξύ: Ανάπτυξη ειδικών συνθετικών παραγώγων και εξειδικευμένων πολυκλωνικών αντισωμάτων

Postgraduate Thesis uoadl:1318185 493 Read counter

Unit:
Κατεύθυνση Βιοχημεία
Library of the School of Science
Deposit date:
2012-07-12
Year:
2012
Author:
Γούρμα Γεωργία
Supervisors info:
Αθανασία Σιαφάκα Καθηγ.(Επιβλέπουσα), Μαίρη Μαυρή-Βαβαγιάννη Αναπλ. Καθηγ., Ευαγγελία Λιβανίου Ερευνήτρια Α΄
Original Title:
Κυανουρικό οξύ: Ανάπτυξη ειδικών συνθετικών παραγώγων και εξειδικευμένων πολυκλωνικών αντισωμάτων
Languages:
Greek
Summary:
Cyanuric acid is an organic molecule of low molecular weight (129 g/mol), which
is widely used as an ingredient of various household and industrial detergents
and as a disinfectant of swimming pool water as well as drinking water.
Moreover, it occurs as a by-product of the industrial synthesis of melamine, a
substance widely used for the production of common polymers. Cyanuric acid has
been found in soil samples after microbial degradation of triazines used as
fertilizers or pesticides and also in marine organisms as a bioaccumulation
product. In addition, in vitro studies with DNA have shown the presence of
cyanuric acid, linked to deoxyribose, as a further oxidation product of
8-oxo-deoxy-guanosine. Intake of cyanuric acid in combination with melamine may
lead to the formation of extremely insoluble crystals with consequent severe
health problems, such as formation of kidney stones, renal failure or even
death. In 2008 a food safety scandal burst out in China, when many newborns who
consumed milk products adulterated with melamine went to hospital and some
died. In view of the importance of cyanuric acid detection, particularly in
food samples, the aim of the present study was to develop, for the first time,
antibodies for cyanuric acid, which could be potentially used in an
immunochemical technique for the analysis of cyanuric acid. Due to its low
molecular weight, cyanuric acid is considered to be a hapten; thus, it should
be conjugated to a suitable immunogenic carrier before administered to a
host-animal for antibody development. For this reason, proper derivatives of
cyanuric acid were designed and synthesized following a protocol based on Fmoc
solid phase peptide synthesis. Then, the synthetic derivatives were purified
with semi-preparative RP-HPLC, characterized with analytical RP-HPLC and
ESI-MS, conjugated to the carrier-protein bovine thyroglobulin and the
conjugates were administered to New Zealand white rabbits. The antibodies
developed were evaluated in an ELISA system, on plates coated first with
streptavidin and then with a biotinylated derivative of cyanuric acid (which
was also synthesized within the frame of the present study), in terms of their
title, their ability to recognize free, parental cyanuric acid in solution and
possible cross-reactivity with structurally related substances, namely
melamine, atrazine and 1,3,5-triazine. The above antibodies may be further
applied to specific immunochemical assays for studies of cyanuric acid.
Keywords:
Cyanuric acid, Melamine, Peptide derivatives of cyanuric acid, Anti-hapten antibodies, ELISA techniques/methods
Index:
Yes
Number of index pages:
1-7
Contains images:
Yes
Number of references:
84
Number of pages:
137

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