Κατεύθυνση Κοσμητολογία-Δερματοφαρμακολογία
Library of the School of Science

2020-10-26

2020

Pappou Charikleia-Maria

Πέτρου Παναγιώτα, Ερευνήτρια Α΄ του Ε.Κ.Ε.Φ.Ε. “ΔΗΜΟΚΡΙΤΟΣ”
Καραλής Ευάγγελος, Επίκουρος Καθηγητής, Τμήμα Φαρμακευτικής, Πανεπιστήμιο Αθηνών
Ράλλης Μιχαήλ, Επίκουρος Καθηγητής, Τμήμα Φαρμακευτικής, Πανεπιστήμιο Αθηνών

ΑΝΕΠΑΡΚΕΙΑ ΒΙΟΤΙΝΗΣ ΚΑΙ ΠΑΘΗΣΕΙΣ ΤΟΥ ΔΕΡΜΑΤΟΣ: ΕΝΖΥΜΙΚΕΣ ΔΕΣΜΕΥΤΙΚΕΣ ΜΕΘΟΔΟΙ ΠΡΟΣΔΙΟΡΙΣΜΟΥ ΤΩΝ ΕΠΙΠΕΔΩΝ ΒΙΟΤΙΝΗΣ ΣΤΟΝ ΑΝΘΡΩΠΙΝΟ ΟΡΟ

Greek

BIOTIN DEFICIENCY AND SKIN DISEASES: ENZYME BINDING METHODS FOR THE DETERMINATION OF BIOTIN’S LEVELS IN HUMAN SERUM

Biotin (vitamin B7) is a water-soluble vitamin of the B-vitamin complex, which is involved, both in humans and in other organisms, in a wide range of metabolic processes, mainly related to gluconeogenesis, fatty acid synthesis and catabolism of the branched-chain amino acids. Consequently, the lack of biotin in the human body results in various diseases related to the nervous system and skin, as well as the body development of the body, seriously threatening the health of patients. Therefore, there is a strong need to develop reliable and sensitive analytical methods for the detection and/or quantification of biotin, as it is important to monitor serum biotin levels in pregnant women, children, people with diabetes, etc. In this frame, the aim of the present thesis was to develop an enzymatic competitive type method for biotin determination in microtiter wells. For this purpose, at first, the following reagents have been tested as binders: a goat polyclonal anti-biotin antibody, a rabbit polyclonal anti-biotin antibody, and streptavidin labelled with horse radish peroxidase. It was found that amongst these three binders only streptavidin provided the necessary sensitivity for biotin levels determination in human serum. After that all the assay parameters were optimoized taking into account both the absolute analytical signal and the assay sensitivity. The method developed was characherized by short duration (1 h), high sensitivity and accuracy, with detection limit comparable or lower to those reported in literature. In particular, the limit of detection for biotin in human serum was 0.5 pmol/mL, allowing the determination of biotin levels in human serum were the values ranged between 0.78 and 3.43 pmol/mL. The analytical capabilities have been demonstrated by analysing human serum samples. In addition, the successful application of the method for the determination of biotin levels in commercially available vitamin supplements was demonstrated.

Science

Health Sciences

biotin, anti-biotin antibody, streptavidin, competitive assay, human blood serum

No

0

No

107

136

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https://pergamos.lib.uoa.gr/uoa/dl/object/2926057