Homologous ELISA for detection of prednisolone in human serum

Food and Agricultural Immunology

T. G. Shrivastav¹  Dinesh Kumar²  Harpal Singh³ 

[1] Immuno, Isotope and Nano-technology Laboratory, Department of Reproductive Biomedicine, The National Institute of Health and Family Welfare (NIHFW), New Delhi, Indi;Immuno, Isotope and Nano-technology Laboratory, Department of Reproductive Biomedicine, The National Institute of Health and Family Welfare (NIHFW), New Delhi, Indi;Indian Institute of Technology Delhi (IIT-D), New Delhi, Indi;All India Institute of Medical Sciences Delhi (AIIMS-D), New Delhi, Indi;Indian Institute of Technology Delhi (IIT-D), New Delhi, Indi;All India Institute of Medical Sciences Delhi (AIIMS-D), New Delhi, Indi;

关键词: Prednisolone;    immunogen;    antibody;    horseradish peroxidase;    ELISA;   

DOI  :  10.1080/09540105.2017.1376184

来源: publisher

【 摘 要 】

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.

【 授权许可】

CC BY