Enzyme Immunoassays for the Determination of Ovine LH and FSH

Abstract

The development of competitive enzyme immunoassays for ovine plasma LH (oLH) and FSH (oFSH) is described. Standards and plasma samples were preincubated with diluted antiserum to oLH or oFSH and the reacted solution (100 micro l per well) was transferred to plates previously coated with oLH or oFSH, respectively. The second antibody used was anti-rabbit IgG horseradish peroxidase. The measuring range was 0.39-50 ng/ml for each hormone and the 50% relative binding sensitivity was 9 ng/ml for oLH. The respective value for oFSH was 3.5 or 34 ng/ml with different hormone and antibody preparations used for the assay. The enzyme immunoassays were used to determine oLH and oFSH levels in plasma from ewes of two breeds during the oestrous cycle. The assays detected the first FSH surge coincident with the LH surge, the second FSH surge about 24 h later and the periodic fluctuations of FSH concentrations during the luteal phase of the oestrous cycle. These enzyme immunoassays are an efficient and economic alternative to the established radioimmunoassays (RIA) for oLH and oFSH.

Development of a Simple Enzyme Immunoassay for the Determination of Ovine Luteinizing Hormone

“…However, similar coefficients have also been described using different antibodies (Peclaris et al, 2003;Yildiz et al, 2003). A higher intra-assay coefficient was found when analysing samples with low concentration of LH, according to the results obtained by Illera and colleagues (1996) when determining rat LH.…”

“…Various EIA methods for the quantification of ovine LH have previously been developed (Spearow and Trost, 1987;Maurel, 1991;Dupont et al, 2003;Peclaris et al, 2003;Yildiz et al, 2003). The first three to be developed were based on the coating of plates with an anti-o LH antibody.…”

Hexagonally Packed Macroporous Molecularly Imprinted Polymers for Chemosensing of Follicle-Stimulating Hormone Protein

“…8 Enzyme-linked immunoassays are another alternative method reported for FSH analysis. 9,10 However, this technique is a multistep one, and requires a long time. 11 Therefore, it is not suitable for point-of-care diagnostics.…”

“…10.1021/acssensors.9b01878. Instruments and technical details of the experiment, SEM images of colloidal crystals, capacitance current measurement at macroporous and thin MIP films, reproducibility test, real sample measurement, and summary of XPS analysis (PDF) E-mail: mcieplak@ichf.edu.pl (M.C.).…”

“…Moreover, this method was capable of identifying the dissociated subunits that might be present in a pharmaceutical preparation as product-related impurities . Enzyme-linked immunoassays are another alternative method reported for FSH analysis. , However, this technique is a multistep one, and requires a long time . Therefore, it is not suitable for point-of-care diagnostics.…”