Kinetic determination of atrazine in foods based on stopped-flow fluorescence polarization immunoassay

Sendra, B; Panadero, S.; Eremin, Sergei A.; Gómez‐Hens, A.

doi:10.1016/s0039-9140(98)00064-2

Cited by 12 publications

(6 citation statements)

References 18 publications

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“…Tracers of 2,4D with different bridge lengths (2, 4, or 6 carbon atoms) have been compared using the same antibody. The most sensitive assay was obtained using the ethylenediamine (2 carbon atom chemical bridge) based tracer. , Similar findings have been reported for atrazine FPIA. , Therefore these kinds of tracers have been used for FPIA optimization. ,, …”

supporting

confidence: 72%

“…8 This is particularly important for residue analysis where the limits of detection (LODs) are very low and the sample matrixes are complex. Efforts for further FPIA optimization are directed toward improvement in sensitivity with the use of new more suitable materials and fluorescent dyes, 13,14 new instruments, 8 and new and alternative methodologies such as 2-PEFA (2-photon excited fluorescence anisotropy) 15 or stoppedflow FPIA, 16,17 where the matrix effect is minimized. The specific antibody used is one of the key components of FPIA, like in any other immunoassay, that determines the quality of the method.…”

mentioning

confidence: 99%

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Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

et al. 2002

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Fluorescence polarization immunoassay (FPIA) is a convenient homogeneous assay, the use of which is restricted in environmental analysis by low sensitivity and matrix effects. We selected the herbicides 2,4D and 2,4,5T to synthesize new L-lysine-based fluorescent tracers using solid-phase chemistry. In addition, three different immunogens of 2,4,5T were prepared for immunization and antibody production. The new tracers and antibodies were adapted to FPIA. Tracers with the hapten attached to the alpha-aminogroup of L-lysine and fluorescein to the e-amino group exhibited at least a 5-fold increased sensitivity when compared to the previously reported ethylenediamine-based tracer (2,4D-EDA-F). The isomeric structure (hapten attached to the e-amino and fluorescein to the alpha-amino group) appeared 7.6 times less sensitive, and all other alternative structures exhibited even lower sensitivities. This observation was confirmed against the monoclonal anti-2,4D antibody E2/G2 and polyclonal anti-2,4,5T antibodies. The affinity constant of 2,4D-EDA-F with E2/G2 was 8.1 times higher when compared with the new tracer, suggesting the more specific nature of the L-lysine-based tracer, the use of which leads to a more sensitive assay. This type of tracer could improve performance and lower substantially the detection limits of FPIAs.

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supporting

confidence: 72%

mentioning

confidence: 99%

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

et al. 2002

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“…In addition, the SF apparatus can track Förster Resonance Energy Transfer, F(t) FRET , when an appropriated dye-pair is attached to each of the reactants or in the same molecule when conformational changes exist during binding [11]. In the p(t) case, it has been used to detect drugs of abuse [12][13][14], and the pesticide atrazine [15]. However, the p(t) signal is not a fundamental function and can lead to calculation errors; in contrast, the r(t) is a fundamental expression that is normalized to the total F(t) signal [16].…”

Section: Introductionmentioning

confidence: 99%

Dual-Channel Stopped-Flow Apparatus for Simultaneous Fluorescence, Anisotropy, and FRET Kinetic Data Acquisition for Binary and Ternary Biological Complexes

et al. 2020

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The Stopped-Flow apparatus (SF) tracks molecular events by mixing the reactants in sub-millisecond regimes. The reaction of intrinsically or extrinsically labeled biomolecules can be monitored by recording the fluorescence, F(t), anisotropy, r(t), polarization, p(t), or FRET, F(t)FRET, traces at nanomolar concentrations. These kinetic measurements are critical to elucidate reaction mechanisms, structural information, and even thermodynamics. In a single detector SF, or L-configuration, the r(t), p(t), and F(t) traces are acquired by switching the orientation of the emission polarizer to collect the IVV and IVH signals however it requires two-shot experiments. In a two-detector SF, or T-configuration, these traces are collected in a single-shot experiment, but it increases the apparatus’ complexity and price. Herein, we present a single-detector dual-channel SF to obtain the F(t) and r(t) traces simultaneously, in which a photo-elastic modulator oscillates by 90° the excitation light plane at a 50 kHz frequency, and the emission signal is processed by a set of electronic filters that split it into the r(t) and F(t) analog signals that are digitized and stored into separated spreadsheets by a custom-tailored instrument control software. We evaluated the association kinetics of binary and ternary biological complexes acquired with our dual-channel SF and the traditional methods; such as a single polarizer at the magic angle to acquire F(t), a set of polarizers to track F(t), and r(t), and by energy transfer quenching, F(t)FRET. Our dual-channel SF economized labeled material and yielded rate constants in excellent agreement with the traditional methods.

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“…[115] είναι µια εναλλακτική τεχνική που χρησιµοποιεί σαν αναλυτική παράµετρο την αρχική ταχύτητα της αντίδρασης αντισώµατος -αντιγόνου, όπως αυτή προσδιορίζεται από τις µεταβολές της πόλωσης φθορισµού στο πρώτο δευτερόλεπτο της αντίδρασης (V P =dP/dt). Η µέθοδος εφαρµόστηκε για τα ζιζανιοκτόνα 2,4D [116] και ατραζίνη [117], σε νερό, χυµό πορτοκαλιού και λευκό κρασί και αποδείχθηκε ότι µε την προσέγγιση αυτή ελαχιστοποιείται το matrix effect και αυξάνεται η ευαισθησία. Τα όρια ανίχνευσης που λήφθηκαν ήταν 4 ng/ml για το 2,4D και 0,2 ng/ml για την ατραζίνη βελτιωµένα τουλάχιστον 5 φορές σε σχέση µε την κλασσική FPIA.…”

Section: γ61 ευαισθησίαunclassified

“…OH (12α) παρουσίασε Ι 50 =93,5 ng/ml δηλαδή µεγαλύτερο κατά τουλάχιστον 7 φορές, ενώ µεγαλύτερο κατά 5 φορές ήταν το Ι 50 του 2,4D-EDA-F (65 ng/ml), που µέχρι τώρα εθεωρείτο ο πιο κατάλληλος για την ανάπτυξη τέτοιου είδους µεθόδων µέτρησης[96,99,103,117]. Το υψηλό Ι 50 του 12α µπορεί εύκολα να εξηγηθεί λαµβανοµένης υπ' όψιν της πλήρους οµολογίας της δοµής του µε το ανοσογόνο όπως φαίνεται στο σχήµα 3.6 Σχήµα 3.6 Σχηµατική αναπαράσταση της δοµικής οµολογίας που εµφανίζεται µεταξύ του ανοσογόνου και των ανιχνευτών 12α,β και της δοµικής ετερολογίας µεταξύ του ανοσογόνου και των ανιχνευτών 13α,β.…”

unclassified

Ανάπτυξη Ανοσοδιαγνωστκών Μεθόδων Πόλωσης Φθορισμού Για Τον Προσδιοριμό Τοξικών Ουσιών

Χατζηδάκης¹

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Για την προστασία των πνευµατικών δικαιωµάτων το περιεχόµενο του Ειδικού Μέρους της διδακτορικής διατριβής επιτρέπεται να αναδηµοσιευτεί µόνο µε τη γραπτή άδεια του επιβλέποντα Καθηγητή. V 9. ∆HMOΣIEYΣEIΣ 1. Tavernarakis N, Hatzidakis G, Krambovitis E. Simple and rapid amplification and detection of nucleic acids. US Patent: serial no. 5,569,582 (1996) 2. Hatzidakis G, Stefanakis A, Krambovitis E. Comparison of different antibody-conjugate derivatives for the development of a sensitive and specific progesterone assay. Journal of Reproduction and Fertility, (1993) 97: 557-561. 3. Hatzidakis G, Katrakili N, Krambovitis E. Development of a direct and specific enzyme immunoassay for the measurement of oestrone sulfate in bovine milk. Journal of Reproduction and Fertility, (1993) 98: 235-240. 4. Stefanakis A, Hatzidakis G, Krambovitis E. Development of a simple and reliable immunoenzymatic technique for the determination of the concentration of progesterone in the milk and serum.

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Kinetic determination of atrazine in foods based on stopped-flow fluorescence polarization immunoassay

Cited by 12 publications

References 18 publications

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

Dual-Channel Stopped-Flow Apparatus for Simultaneous Fluorescence, Anisotropy, and FRET Kinetic Data Acquisition for Binary and Ternary Biological Complexes

Ανάπτυξη Ανοσοδιαγνωστκών Μεθόδων Πόλωσης Φθορισμού Για Τον Προσδιοριμό Τοξικών Ουσιών

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