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[Preparation and identification of monoclonal antibody against human spermine oxidase]
Chun-hong Qinli1, Jian-lin Yang, Yu Han
1Institute of Molecular Biology, Medical College, Three Gorges University, Yichang 443002, China.
Aim:
To prepare mouse anti-human spermine oxidase (anti-hSMO) monoclonal antibody (mAb) and testify its application in the biological techniques including Western blotting and immunohistochemistry.
Methods:
Plasmid pET-15b/SMO was first transferred into BL21 (DE3), and then SMO recombinant protein with 6×His tag was induced to express by IPTG and purified by Ni-NTA resin. The purified recombinant SMO was used to immunize BALB/c mouse. The spleen cells from the immunized mouse were harvested and hybridized with Sp2/0 myeloma cells to obtain a hybridoma cell line that could efficiently synthesize and secret anti-SMO mAb. The titer and antigen specificity of this antibody were identified by ELISA, Western blotting and immunohistochemistry.
Results:
We successfully obtained the hybridoma cell line which could stably secret anti-SMO mAb. The mAb was of a high titer and antigen specificity and could be used in ELISA, Western blotting, and immunohistochemistry for SMO.
Conclusion:
The mouse anti-hSMO mAB with a high antigen specificity has been prepared successfully and used for a variety of bio-analytical techniques.
