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Published on: January 25, 2018
A sensitive sandwich ELISA for buffalo prolactin.
Manoj Panchal1, K Muralidhar, S K Gupta
1Hormone Research Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Journal of Immunoassay & Immunochemistry
|April 12, 2007
Summary
Researchers developed a sensitive sandwich ELISA assay for buffalo prolactin (buPRL) using monoclonal antibodies. This assay demonstrates high precision and parallelism with buffalo serum samples, validating its use.
Area of Science:
- Immunology
- Biochemistry
- Veterinary Science
Background:
- Buffalo prolactin (buPRL) is a crucial hormone in buffalo reproduction and lactation.
- Accurate quantification of buPRL is essential for reproductive and physiological studies in buffalo.
- Existing methods for buPRL detection may lack sensitivity or require complex procedures.
Purpose of the Study:
- To develop and validate a sensitive sandwich ELISA (sELISA) for the quantification of buffalo prolactin (buPRL).
- To assess the feasibility of using murine monoclonal antibodies (MAbs) in a direct competitive immunoassay format.
Main Methods:
- Development of two murine monoclonal antibodies (MAbs), designated 1501 and 1504, against buffalo prolactin.
- Establishment of a sandwich ELISA (sELISA) using the 1504 MAb as the capture antibody and anti-buPRL rabbit serum as the detection antibody.
- Evaluation of assay sensitivity, parallelism with buffalo serum samples, and intra- and inter-assay coefficients of variance.
Main Results:
- A functional sELISA system was successfully established using the 1504 MAb and anti-buPRL rabbit serum.
- The developed sELISA exhibited a high sensitivity of 156 pg/mL for buPRL detection.
- Buffalo serum samples demonstrated parallelism with the standard curve, indicating assay specificity.
- The assay showed low variability, with intra-assay and inter-assay coefficients of variance of 8.4% and 9.06%, respectively.
Conclusions:
- The developed sandwich ELISA is a valid and sensitive method for quantifying buffalo prolactin (buPRL).
- The assay's performance characteristics support its utility in research and diagnostic applications involving buffalo endocrinology.
- The use of specific monoclonal antibodies contributes to the assay's reliability and precision.
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