title: [Role of molecular adjuvants TBhsp and MT in the preparation of monoclonal antibody against phosphatase of regenerating liver-3].
abstract: OBJECTIVE
  To explore the adjuvant effects of Mycobacterium tuberculosis heat shock protein (TBhsp) and Mycobacterium tuberculosis T cell stimulatory epitope (MT) in the preparation of the monoclonal antibody (mAb) against phosphatase of regenerating liver-3 (PRL-3).
  
  
  METHODS
  The prokaryotic expression vectors pET28a were used for the construction of pET28a-PRL-3 (control plasmids), pET28a-PRL-3-MT, pET28a-TBhsp-PRL-3 and pET28a- TBhsp-PRL-3-MT (recombinant plasmids). The various fusion proteins expressed in bacteria were purified and utilized to immunize the BALB/c mice, respectively. The serum level of specific anti-PRL-3 antibody was measured by ELISA. The mouse with the highest serum level of anti-PRL-3 antibody was selected to prepare mAb with hybridoma technique. The subclasses of mAb were identified.
  
  
  RESULTS
  The PRL-3 fusion proteins expressed from the four expression plasmids were purified successfully. The serum level of anti-PRL-3 antibody from the mice immunized with PRL-3-MT was the highest compared with the other immune groups. Ten hybridoma cell lines secreting anti-PRL-3 mAb were obtained after cell fusion and ELISA primary screening, and all of the mAb subclasses were IgG.
  
  
  CONCLUSIONS
  MT is a potentially effective molecular adjuvant in preparation of mAb specific for PRL-3.
authors.affiliation: Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Peking University Cancer Hospital &amp; Beijing Institute for Cancer Research, Beijing 100142; Department of Biochemistry, Changzhi Medical College, Changzhi 046000, China.
authors.forename: Yanfei
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Beijing Institute for Cancer Research, Beijing 100142; Department of Clinical Laboratory, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Lv
authors.affiliation: Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Beijing Institute for Cancer Research, Beijing 100142, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Meng
authors.affiliation: Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Beijing Institute for Cancer Research, Beijing 100142, China.
authors.forename: Like
authors.initials: L
authors.lastname: Qu
authors.affiliation: Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Beijing Institute for Cancer Research, Beijing 100142, China.
authors.forename: Chengchao
authors.initials: C
authors.lastname: Shou
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007114:Immunization; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D018984:Epitopes, T-Lymphocyte; D006360:Heat-Shock Proteins; D017874:Immediate-Early Proteins; D007074:Immunoglobulin G; C111534:Ptp4a3 protein, mouse; D011993:Recombinant Fusion Proteins; D017027:Protein Tyrosine Phosphatases