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[Preparation and identification of mouse anti-human CD52 functional antibody]
1Institute of Hematology & Hospital of Hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China. liuyou2k@163.com
Objective:
To prepare and identify mouse anti-human CD52 antibody.
Methods:
The RNA was extracted from Hut-78 cells. CD52 gene was amplified by RT-PCR. With the double-enzyme digestion, CD52 gene was cloned into pcDNA3.1(+) eukaryotic expression vector, named as pcDNA3.1(+)/CD52. We transfected the recombinant into CHO cell by Lipofectamine™ 2000. Stable transfected CHO cell line was established, and the CD52 expression in the transfected cells was detected by RT-PCR, FACS and immunocytochemical staining. BALB/c mice were immunized with synthetic peptide CD52. The titer of anti-CD52 serum was detected with indirect ELISA. Inhibition of leukemia LCL cell proliferation by immunized mouse serum was identified with MTS assay.
Results:
The eukaryotic expression vector pcDNA3.1(+)/CD52 was constructed, stable transfected CHO cell line was established. High titer anti-CD52 antibody was identified. The immunized mouse serum inhibits proliferation of LCL cells by MTS assay.
Conclusion:
Preparation of functional antibody against CD52 and stable transfected CHO cell line have established successfully.
