title: [Preparation and identification of mouse anti-human CD52 functional antibody].
abstract: 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.
  
  
  CONCLUSIONS
  Preparation of functional antibody against CD52 and stable transfected CHO cell line have established successfully.
authors.affiliation: Institute of Hematology & Hospital of Hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China. liuyou2k@163.com
authors.forename: You
authors.initials: Y
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Qu
authors.forename: Lin
authors.initials: L
authors.lastname: Shi
authors.forename: Lihua
authors.initials: L
authors.lastname: Huang
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Weibin
authors.initials: W
authors.lastname: Zhu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D000074301:CD52 Antigen; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D051379:Mice
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; C000613862:CD52 protein, human; D018076:DNA, Complementary; D006023:Glycoproteins