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This Article Full Text Full Text (PDF) An addition or correction has been published All Versions of this Article: 72/1/78    most recent biolreprod.104.033654v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jimenez, M. Articles by Handelsman, D.J. Search for Related Content PubMed PubMed Citation Articles by Jimenez, M. Articles by Handelsman, D.J. Agricola Articles by Jimenez, M. Articles by Handelsman, D.J.

Validation of an Ultrasensitive and Specific Immunofluorometric Assay for Mouse Follicle-Stimulating Hormone¹

Andrology Laboratory, ³ ANZAC Research Institute & Concord Hospital, University of Sydney, Sydney, New South Wales 2139, Australia NV Organon,⁴ 5342 Oss, The Netherlands

Sensitive and specific measurement of FSH is critical to research in reproductive biology, and the increasing availability of transgenic mouse models has created a need for a robust, sensitive, and specific mouse (m) FSH assay. The present study evaluated a time-resolved immunofluorometric assay (IFMA) for mFSH using monoclonal antibody to human (h) FSHß as a capture antibody and a biotinylated polyclonal antibody to rat subunit as a detection probe, with signaling amplified by europium-labeled streptavidin. The mFSH IFMA lowered the detection limit 34-fold (5 vs. 170 pg/sample) compared with standard mFSH RIA. The mFSH IFMA demonstrated parallelism of response to dilutions of castrated mouse serum and rat FSH but no cross-reactivity with hFSH and mLH or hLH, whereas the RIA demonstrated nonparallel cross-reactivity with hFSH. The IFMA has a wide analytical range, with a good precision profile for within- and between-assay reproducibility. Because the IFMA is a sandwich-type assay with strict dimer-specificity by design, the lower readings and recovery obtained were compared with the RIA when both assays used a pituitary-purified mFSH assay standard that contained isolated or fragmented subunits as well as intact dimeric FSH. When used with mouse serum sample, the mFSH IFMA demonstrated the expected increases following orchidectomy as well as markedly enhanced sensitivity to very low levels of endogenous mFSH in gonadotropin-deficient mice. Furthermore, the IFMA measured mFSH with fidelity in both intact and orchidectomized male mice without any interference from transgenic hFSH. The greatly enhanced sensitivity, specificity, and technical convenience of this mFSH IFMA will allow wider application of FSH measurements to very small blood samples in immature and mature mice as well as transgenic models.

¹ Supported by the NHMRC.

² Correspondence: D.J. Handelsman, ANZAC Research Institute, Sydney NSW 2139, Australia. Fax: 61 29 767 9101; djh{at}anzac.edu.au

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