Journal
MOLECULAR MEDICINE REPORTS
Volume 15, Issue 6, Pages 4035-4040Publisher
SPANDIDOS PUBL LTD
DOI: 10.3892/mmr.2017.6523
Keywords
soluble CD147; polyclonal antibody; purification; immunoaffinity chromatography; sandwich ELISA
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Funding
- National Science and Technology Major Project [2012AA02A301]
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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 similar to 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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