Open Access

Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography

  • Authors:
    • Shuangshuang Liu
    • Shasha Li
    • Yang Zhang
    • Ye Wang
    • Yumeng Zhu
    • Bin Wang
    • Zhi‑Nan Chen
  • View Affiliations

  • Published online on: April 27, 2017     https://doi.org/10.3892/mmr.2017.6523
  • Pages: 4035-4040
  • Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The immunoglobulin superfamily member CD147 is a widely expressed glycoprotein that occurs in both a membrane‑spanning and soluble form. Sandwich ELISA is a powerful tool for analyzing soluble antigens. The aim of the present study was to obtain a highly specific polyclonal antibody against human CD147 that can be used for sandwich ELISA analysis. Expression of recombinant CD147 by a eukaryotic expression system was used to immunize rabbits to obtain antiserum. A highly specific polyclonal antibody that was able to detect soluble CD147 in sandwich ELISA was obtained by antigen‑immunoaffinity chromatography purification. The purity of rabbit anti‑CD147 polyclonal antibodies was ~99%, and ELISA analysis was able to determine the titer of the rabbit anti‑CD147 polyclonal antibodies at 1:512,000. The lowest concentration of the standard CD147 antigen that the sandwich ELISA was able to detect was 31.25 pg/ml. The sandwich ELISA system was composed of anti‑hepatoma HAb18 monoclonal antibodies and purified rabbit anti‑CD147 polyclonal antibodies. The present study demonstrated that antigen‑immunoaffinity chromatography may be a good technique for the purification of polyclonal antibodies, which may be used to detect antigen in sandwich ELISAs.
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June-2017
Volume 15 Issue 6

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Liu S, Li S, Zhang Y, Wang Y, Zhu Y, Wang B and Chen ZN: Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Mol Med Rep 15: 4035-4040, 2017
APA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., & Chen, Z. (2017). Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography. Molecular Medicine Reports, 15, 4035-4040. https://doi.org/10.3892/mmr.2017.6523
MLA
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15.6 (2017): 4035-4040.
Chicago
Liu, S., Li, S., Zhang, Y., Wang, Y., Zhu, Y., Wang, B., Chen, Z."Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography". Molecular Medicine Reports 15, no. 6 (2017): 4035-4040. https://doi.org/10.3892/mmr.2017.6523