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  • Title: Purification of a polyclonal antibody against CD147 for ELISA using antigen‑immunoaffinity chromatography.
    Author: Liu S, Li S, Zhang Y, Wang Y, Zhu Y, Wang B, Chen ZN.
    Journal: Mol Med Rep; 2017 Jun; 15(6):4035-4040. PubMed ID: 28487989.
    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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