Determination of prostaglandin metabolites in biological samples by competitive time-resolved fluoroimmunoassay

Lüke, Franz J.; Schlegel, Werner

doi:10.1016/0022-1759(92)90175-s

Cited by 4 publications

(2 citation statements)

References 15 publications

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“…In recent years, various other methods have gradually been replacing RIA. Among them, time-resolved fluoroimmunoassay (TR-FIA), using the long-lived fluorescence of chelates of lanthanides such as europium (Eu) as the label (Hemmilä et al, 1984;Lövgren et al, 1985;Hemmilä and Harju, 1994), has become available for measuring various hormones (LUke and Schlegel, 1992;Ditkoff et al, 1993;lus et al, 1993;Iwasawa et al, 1994;Yamada et al, 1997).…”

Section: Endocrinology Anddiabetesmentioning

confidence: 99%

Time-resolved fluoroimmunoassay (TR-FIA) of porcine relaxin

Ogine¹,

Kohsaka²,

Taya³

2009

Exp Clin Endocrinol Diabetes

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We developed and validated a new assay system for porcine relaxin that overcame the drawbacks of RIA by adapting time-resolved fluoroimmunoassay (TR-FIA), which was recently introduced as a non-RIA format. The assay system was a solid-phase TR-FIA based on competition for a polyclonal anti-porcine relaxin antibody between europium (Eu)-labeled porcine relaxin and test samples. Antibody-relaxin complexes were then bound to the second antibody coated on the solid phase, achieving rapid and complete separation of bound and free antigen. A standard curve was produced over the range of 1 pg/well to 1000 pg/well. Serum and corpus luteum extracts from pigs in late pregnancy exhibited inhibition curves parallel to that of the relaxin standard, whereas male pig serum caused no displacement of the labeled hormone. No cross-reactivity was seen with other hormones, such as insulin, LH, and FSH, indicating a high specificity of the assay. The sensitivity was 4 pg/well (80 pg/ml), which was high and equivalent to that of the porcine relaxin RIA. The intra-assay and inter-assay coefficients of variation were less than 3.8% and 6.7%, respectively. Recovery of porcine relaxin added to male pig serum sample averaged 103%. The advantages of this TR-FIA were that addition of tyrosine was not necessary for labeling, unlike the RIA, Eu-labeled relaxin was stable enough to allow long-term storage for more than one year, the assay was completed in only 5 h versus two to seven days for the RIA, and no special safety precautions were needed. To validate this TR-FIA, the serum relaxin concentrations during late pregnancy, parturition and early lactation were investigated in pigs. Serum relaxin levels determined by this assay were similar to those obtained previously by RIA. In conclusion, this TR-FIA could replace RIA as the method of choice for assay of relaxin.

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Section: Endocrinology Anddiabetesmentioning

confidence: 99%

Time-resolved fluoroimmunoassay (TR-FIA) of porcine relaxin

Ogine¹,

Kohsaka²,

Taya³

2009

Exp Clin Endocrinol Diabetes

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“…Σήμερα, εφαρμόζονται και ανοσολογικές μέθοδοι ποιοτικής και ποσοτικής ανίχνευσης των ισοπροστάνων, κατά τις οποίες ειδικά αντισώματα σχηματίζουν ανιχνεύσιμα ανοσοσυμπλέγματα με τις ισοπροστάνες και ειδικότερα με την 8-isoprostagladin -F 2a . Θεωρούνται όμως χαμηλότερης ειδικότητας λόγω πιθανής διασταυρούμενης αντίδρασης με άλλα προστανοειδή62,67 .Οι ανωτέρω μέθοδοι αποτελούν τις πιο συχνά χρησιμοποιούμενες μεθόδους για την ακριβή ανίχνευση και ποσοτικό προσδιορισμό των Ε.Ρ.Ο σε καταστάσεις ισχαιμίας και επαναιμάτωσης. Σήμερα, εφαρμόζονται και άλλες μέθοδοι που επίσης βασίζονται στην ανίχνευση προϊόντων της υπεροξείδωσης των λιπαρών οξέων και επιτρέπουν την μέτρηση της συνολικής ικανότητας του αίματος να «εκκαθαρίζει» ή να «συλλαμβάνει» Δ.Μ.Ο.…”

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