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Comparison of an in- house Elisa kit with a commercially available kit for the measurement of pregnant mare serum gonadotropin (PMSG)

Nitisha Tyagi, Md. Summon Hossain, Taruna Arora, Kambadur Muralidhar*

Abstract


Antiserum to equine Luteinizing Hormone (eLH) raised in a rabbit was used to standardize a competitive ELISA to measure PMSG in native state. The characteristics of this immunoassay developed in-house were compared with those of a commercially available kit. The sensitivity, intra- assay and inter-assay coefficient of variation of both the assays were comparable. Using the in-house assay, immobilized Cibacron Blue, a textile dye, was shown to bind PMSG selectively PMSG. This enabled isolation of PMSG in native condition from pregnant mare serum as well as from enriched serum fractions. It was also observed that a commercial preparation of PMSG gives underestimates when assayed in the commercial kit. Hence there is no correlation between bioassay Units and Immunoassay units when estimated using different antibodies strongly indicating existence of different conformers/isoforms of PMSG.

Keywords


Pregnant mare serum gonadotropin; ELISA; eLH antiserum; monoclonal antibody

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References


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DOI: http://dx.doi.org/10.21746/ijbio.2015.10.0019

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