The aim of this research was to develop an assay that can detect the cancer biomarker Thomsen-nouvelle (Tn) antigen using ELISA. The study investigated the impact of the size and functional groups of magnetic beads (MBs) on the specific sensitivity of the bioaffinity interaction. We used four different carboxy-modified MBs with varying sizes (250 nm, 500 nm, 1000 nm, and 2800 nm) to evaluate the best MBs for detecting anti-Tn antibodies. We calculated the slopes from calibration curves to determine the detection sensitivities, and assessed the effect of size on the specific sensitivity of detection of anti-Tn antibodies. We obtained a limit of detection (LOD) of (0.31 ± 0.01) ng mL− 1 or (2.10 ± 0.04) pM for analyte detection, and the optimal assay configuration was highly selective and allowed for reliable detection of the analyte in human serum with a recovery index in the range of 102 -104%.
This work was supported by the Slovak Research and Development Agency under the Contract no. APVV-21-0329.
[1] Sung H., Ferlay J., Siegel R. L. et al. (2021) CA: CA Cancer J Clin. 71(3), p. 209.
[2] Mani V., Chikkaveeraiah B.V., Rusling J.F. (2011) Expert Opin. Med. 5(5), p. 381.
[3] Blšákova A., Květoň F., Lorencová L. et al. (2022) Anal. Chim. Acta. 1195: p. 339444.
[4] Vráblová V., Blšákova A., Lorencová L. et al. (2023) Anal. Chim. Acta. 1242: p. 340794.
Impressive data,
Impressive data, congratulations to your results! The crucial step for any diagnostic method is the appropriate preparation of analytical sample. Thusly, I would like to know how do you deal with your biological inputs (serum? plasma? body fluids?), how do you purificate anti-glycan antibodies (AGA) matrix to be satisfyingly accessible for the proposed detection method. Thank you in advance for your remarks, ZB
Thank you very much. I
Thank you very much. I appreciate your question and the initiation of this discussion.
Indeed, we have established a collaboration with an oncology physician who supplies us with serum samples obtained from cancer patients. Nevertheless, for the sake of ensuring reproducibility in our measurements, we also utilize commercially available healthy serum from Merck. This serum undergoes thorough analysis and is accompanied by fundamental data. By incorporating the purchased sera into our study, we can maintain consistency while optimizing our methodology. The undeniable advantage of using magnetic particles with a specific surface treatment is their affinity for AGA and the possibility of detection without pretreatment of the sample.
If you have any further questions, please feel free to reach out to me.
Ok, I see... thank you very
Ok, I see... thank you very much for clarifying conducting your procedures. Wish you all the best, keep doing great job! Sincerely, ZB.
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