title: [Expression and polyclonal antibody preparation of HPV-11E7 protein].
abstract: OBJECTIVE
  To express human papillomavirus type 11, E7 protein (HPV11E7) via a prokaryotic expression vector and produce anti-HPV11E7 polyclonal antibody.
  
  
  METHODS
  A prokaryotic expression vector pGEX-4T2-HPV11E7 was constructed and soluble GST-HPV11E7 fusion protein was expressed in E.coli by IPTG induction and purified. The purified HPV11E7 protein was used to immunize New Zealand rabbits to prepare the anti-HPV11E7 polyclonal antibody followed by protein G agarose purification to obtain the IgG type polyclonal antibody. Western blotting and immunofluorescence analysis were used to test the specificity and titer of the antibody.
  
  
  RESULTS
  SDS-PAGE analysis demonstrated that large amounts of soluble GST-HPV11E7 fusion protein was expressed in E.coli after 6 hours of IPTG induction. Western blotting and immunofluorescence confirmed that the purified anti-HPV11E7 polyclonal IgG antibody obtained from immunized rabbits had a high titer and specificity.
  
  
  CONCLUSIONS
  The prokaryotic expression system could express a great deal of soluble HPV11E7 protein, and anti-HPV11E7 polyclonal IgG antibody from HPV11E7-immunized rabbits was proved to have a high titer and specificity.
authors.affiliation: Department of Dermatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Center of Biomedicine Research, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
authors.forename: Shaojie
authors.initials: S
authors.lastname: Jiang
authors.affiliation: Department of Dermatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
authors.forename: Xianzhen
authors.initials: X
authors.lastname: Chen
authors.affiliation: Center of Clinical Medicine Research, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Chen
authors.affiliation: Center of Clinical Medicine Research, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Dermatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D052140:Human papillomavirus 11 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology*; D008856:Microscopy, Fluorescence; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D050725:Papillomavirus E7 Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase