Preparation and characterization of monoclonal antibodies against mouse Metrnl

QU Yi; LI Zhiyong; MIAO Chaoyu

doi:10.3969/j.issn.1006-0111.2017.02.007

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Objective To prepare monoclonal antibodies (mAb) against mouse Metrnl and identify its specificity. Methods Mouse Metrnl polypeptide fragments and full-length protein were prepared as antigens to immunize mice.Then mice spleen cells were fused with SP2/0 myeloma cells to obtain hybridoma cells which were screened for positive clone in order to subclone for stable cell lines.After ascites were prepared, ELISA method was used to detect the antibodies titer.Western blot method was applied to identify their specificity. Results No effective antibodies were identified from the ascites derived from 14 polypeptide antigens.Among the 25 antibodies derived from the full-length protein, 12 monoclonal antibodies can be used to identify the recombinant Metrnl protein. Conclusion 12 monoclonal antibodies were successfully prepared to identify mouse Metrnl protein.

Preparation and characterization of monoclonal antibodies against mouse Metrnl

Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China

Received Date: 2016-11-08

Rev Recd Date: 2016-12-23

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Abstract: Objective To prepare monoclonal antibodies (mAb) against mouse Metrnl and identify its specificity. Methods Mouse Metrnl polypeptide fragments and full-length protein were prepared as antigens to immunize mice.Then mice spleen cells were fused with SP2/0 myeloma cells to obtain hybridoma cells which were screened for positive clone in order to subclone for stable cell lines.After ascites were prepared, ELISA method was used to detect the antibodies titer.Western blot method was applied to identify their specificity. Results No effective antibodies were identified from the ascites derived from 14 polypeptide antigens.Among the 25 antibodies derived from the full-length protein, 12 monoclonal antibodies can be used to identify the recombinant Metrnl protein. Conclusion 12 monoclonal antibodies were successfully prepared to identify mouse Metrnl protein.

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Reference (14)

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Preparation and characterization of monoclonal antibodies against mouse Metrnl

doi: 10.3969/j.issn.1006-0111.2017.02.007

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