A Sensitive Competitive ELISA for Determination of Biotin in Transformed Yeast Culture Media

Abstract

Abstract: Aim To develop a sensitive competitive ELISA for the determination of biotin in transformed yeast culture media. Methods The ELISA plate was firstly coated with Mycoplasma hyopneumoniae, and then successively incubated with rabbit anti-Mycoplasma hyopneumoniae serum and goat anti-rabbit IgG-biotin to form the solid biotin, which competed with the biotin in the solution (standard or sample) for the limited streptavidin-horse radish peroxidase conjugate. The stan-dard calibration curve for biotin analysis was constructed in the range of 50-2000 ng·L^-1. Results The detection limit for biotin was found to be 83 ng·L^-1, which was about 1000 times lower than the lowest determination concentration in the re-ported ELISA for biotin analysis. The relative standard deviations for the spiked samples at biotin concentrations of 200 ng·L^-1, 500 ng·L^-1, and 1000 ng·L^-1 were 24.87%, 6.15%, and 7.86%, respectively, with the average recovery of 101.13%. The wild yeast and its sixty-three transformed yeast culture media were applied to the developed ELISA for the determination of biotin. It was found that the biotin concentrations in more than 85 % of the tested samples were enhancedwith different increase factors after transformation. Conclusion Utilization of Mycoplasma hyopneumoniae as the coating protein improves the precision and accuracy of the ELISA assay, which might be used for the biotin assay in other media.

Key words: affinity assay, affinity assay, biotin analysis, biotin analysis, streptavidin, streptavidin, ELISA, ELISA, coating, coating, transformed yeast culture media, transformed yeast culture media, Mycoplasma hyopneumoniae, Mycoplasma hyopneumoniae

CLC Number:

R927.3

Supporting:

YANG Hong^*. A Sensitive Competitive ELISA for Determination of Biotin in Transformed Yeast Culture Media[J]. .

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