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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Foodborne Toxin Detection and Prevention Research » Research » Publications at this Location » Publication #365722
Research Project: Advance the Development of Technologies for Detecting and Determining the Stability and Bioavailability of Toxins that Impact Food Safety and Food Defense

Location: Foodborne Toxin Detection and Prevention Research

Title: Comparison of soybean peroxidase with horseradish peroxidase and alkaline phosphatase used in immunoassays

Author
item YANG, HUIJUAN - China Agriculture University
item Bever, Candace
item ZHANG, HUIYAN - China Agriculture University
item MARI, GHULAM - China Agriculture University
item LI, HONGFANG - China Agriculture University
item ZHANG, XIYA - Henan Agricultural University
item GUO, LIUCHUAN - China Agriculture University
item WANG, ZIWEN - China National Center For Food Safety Risk Assessment
item LUO, PENGJIE - China National Center For Food Safety Risk Assessment
item WANG, ZHANHUI - China Agriculture University

Submitted to: Analytical Biochemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/7/2019
Publication Date: 6/13/2019
Citation: Yang, H., Bever, C.R., Zhang, H., Mari, G.M., Li, H., Zhang, X., Guo, L., Wang, Z., Luo, P., Wang, Z. 2019. Comparison of soybean peroxidase with horseradish peroxidase and alkaline phosphatase used in immunoassays. Analytical Biochemistry. 581:113336. https://doi.org/10.1016/j.ab.2019.06.007.
DOI: https://doi.org/10.1016/j.ab.2019.06.007

Interpretive Summary: Immunoassays are a valuable tool used in food safety. To improve their use, it is essential to select an economical enzyme reporter that possessed high stability and high efficiency. In this research, soybean peroxidase (SBP) was compared with horseradish peroxidase (HRP) and alkaline phosphatase (ALP) used as reporter enzymes in an immunoassay detecting ractopamine, as the model system. The results indicated that SBP, an anionic enzyme isolated from waste soybean hulls, outperformed HRP and ALP in terms of signal stability and its tolerance to the buffer conditions tested, which included a range of pHs, as well as, exposure to organic solvents and heat. SBP-antibody conjugate (SBP-Ab) based immunoassays possessed a lower LOD, better accuracy and precision in the detection of swine, cattle and sheep urine samples. These results suggested that SBP could adequately replace the use of HRP and ALP in immunoassays for the detection of veterinary drug residue and also has great potential in the rapidly-developing immunoassay field.

Technical Abstract: Enzyme labeling of an antigen or an antibody helps to visualize and amplify the signal and is an important reagent used in immunoassays for the detection of a target of interest. In this research, soybean peroxidase (SBP), a less commonly used enzyme reporter, was compared in immunoassays with the two most commonly used reagents, horseradish peroxidase (HRP) and alkaline phosphatase (ALP). The enzyme-antibody conjugates were evaluated by their performance in an indirect competitive enzyme-linked immunosorbent assay icELISA) and in an indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) for ractopamine (RAC). The results revealed that the more affordable SBP offers a long-lasting chemiluminescent signal, which outperformed ALP and HRP. SBP-antibody conjugate (SBP-Ab) based immunoassays produced lower limits of detection (LODs) and better accuracy in the detection of RAC in animal urine samples. Additionally, SBP-Ab has advantages in being more resistant to heat, acid and organic solvents. These results suggest that SBP could be a potentially excellent alternative to HRP and ALP for the development of immunoassay in food safety field.