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Tsai H, Lu YH, Liao HX, et al. Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate. Chem Cent J. 2015;9:8doi: 10.1186/s13065-015-0088-1.
Tsai, H., Lu, Y. H., Liao, H. X., Wu, S. W., Yu, F. Y., & Fuh, C. B. (2015). Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate. Chemistry Central journal, 98. https://doi.org/10.1186/s13065-015-0088-1
Tsai, Hweiyan, et al. "Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate." Chemistry Central journal vol. 9 (2015): 8. doi: https://doi.org/10.1186/s13065-015-0088-1
Tsai H, Lu YH, Liao HX, Wu SW, Yu FY, Fuh CB. Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate. Chem Cent J. 2015 Feb 22;9:8. doi: 10.1186/s13065-015-0088-1. eCollection 2015. PMID: 25745512; PMCID: PMC4350001.
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Chem Cent J. 2015 Feb 22;9:8. doi: 10.1186/s13065-015-0088-1. eCollection 2015.

Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate.

Chemistry Central journal

Hweiyan Tsai, Yi-Hsuan Lu, Huan-Xuan Liao, Shih-Wei Wu, Feng-Yih Yu, Chwan Bor Fuh

Affiliations

  1. School of Medical Applied Chemistry, Chung Shan Medical University, Taichung, 402 Taiwan ; Department of Medical Education, Chung Shan Medical University Hospital, Taichung, 402 Taiwan.
  2. School of Medical Applied Chemistry, Chung Shan Medical University, Taichung, 402 Taiwan.
  3. School of Biomedical Sciences, Chung Shan Medical University, Taichung, 402 Taiwan.
  4. Department of Applied Chemistry, National Chi Nan University, Puli, Nantou, 545 Taiwan.

PMID: 25745512 PMCID: PMC4350001 DOI: 10.1186/s13065-015-0088-1

Abstract

BACKGROUND: The enzyme-linked immunosorbent assay (ELISA) has been used for diagnosing medical and plant pathologies. In addition, it is used for quality-control evaluations in various industries. The ELISA is the simplest method for obtaining excellent results; however, it is time consuming because the immunoreagents interact only on the contact surfaces. Antibody-labeled magnetic particles can be dispersed in a solution to yield a pseudohomogeneous reaction with antigens which improved the efficiency of immunoreaction, and can be easily separated from the unreactive substances by applying a magnetic force. We used a homemade magnetic microplate, functional magnetic particles (MPs) and enzyme-labeled secondary antibody to perform the sandwich ELISA successfully.

RESULTS: Using antibody-labeled MPs enabled reducing the analysis time to one-third of that required in using a conventional ELISA. The secondary antibody conjugated with horseradish peroxidase (HRP) was affinity-bound to the analyte (IgG in this study). The calibration curve was established according to the measured absorbance of the 3, 3', 5, 5'-tetramethybezidine-HRP reaction products versus the concentrations of standard IgG. The linear range of IgG detection was 114 ng/mL-3.5 ng/mL. The limit of detection (LOD) of IgG was 3.4 ng/mL. The recovery and coefficient of variation were 100% (±7%) and 116% (±4%) for the spiked concentrations of 56.8 ng/mL and 14.2 ng/mL, respectively.

CONCLUSION: Pseudohomogeneous reactions can be performed using functional MPs and a magnetic microplate. Using antibody-labeled MPs, the analysis time can be reduced to one-third of that required in using a conventional ELISA. The substrate-enzyme reaction products can be easily transferred to another microplate, and their absorbance can be measured without interference by light scattering caused by magnetic microbeads. This method demonstrates great potential for detecting other biomarkers and in biochemical applications. Graphical AbstractA magnetic ELISA with convenient magnetic microplate.

Keywords: Functional magnetic particles; Immunoassay; Magnetic separator

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