Food and Agricultural Immunology (Jan 2018)

Homologous ELISA for detection of prednisolone in human serum

  • Dinesh Kumar,
  • Harpal Singh,
  • T. G. Shrivastav

DOI
https://doi.org/10.1080/09540105.2017.1376184
Journal volume & issue
Vol. 29, no. 1
pp. 369 – 385

Abstract

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An enzyme-linked immunosorbent assay to measure prednisolone have been developed. An antibody was raised in rabbits using prednisolone-21-hemisuccinate (PSL-21-HS) conjugated to bovine serum albumin. Similarly, PSL-21-HS was conjugated to horseradish peroxidase to prepare enzyme conjugate. The developed assay has been validated for sensitivity and effective displacement at 50% of the assay were 0.078 and 2.64 ng/mL, respectively. This assay showed cross-reaction with only 4 steroids – i.e. progesterone 1.76%, 17α-OH progesterone 5.89%, cortisol 7.69% and prednisone 1.13%, out of 55 analogous steroids. The percent recovery of prednisolone from the exogenously spiked human serum pools was in the range of 94.84–100.17%. The intra-assay and inter-assay coefficients of variation ranged from 5.79% to 8.00% and from 3.23% to 8.63%, respectively. The serum prednisolone values obtained by this method correlated well with the commercially available kit and found to be 0.93.

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