ISSN 2410-7751 (Print)
ISSN 2410-776X (on-line)
"Biotechnologia Acta" V. 9, No 2, 2016
https://doi.org/10.15407/biotech9.02.028
Р. 28-36, Bibliography 33, English
Universal Decimal Classification: 557.161.2+612.017.1:616-097
A. O. Mazanova, I. O. Shymanskyy, M. M. Veliky
Palladin Institute of Biochemistry of NAS of Ukraine, Kyiv
The present study was aimed at designing of immunoenzyme test-system for competitive determination of 25OHD in blood serum using polyclonal antibodies against 25OHD and biotin-streptavidin visualization technique. The validation parameters of test-system were determined. For development of test-system we used the preparation of specific antibodies against 25OHD with the titer 1:3000. The optimal concentration of competing agent (25OHD3-LC-biotin) was found to be equal to 5 ng/ml. The standard calibration curve was built by using a set of seven calibrators. For the validation of test-system the following parameters were established: limit of detection, quantitative limit, cross-reactivity, intra- and inter-system coefficients of variation (Intra.CV, Inter.CV) and “matrix effect”. Constructed immunoenzyme test-system can be used successfully for 25OHD determination in blood serum samples.
Key words: 25-Hydroxyvitamin D3, immunoenzyme analysis, polyclonal antibodies.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2016
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