title: [Purification of Cyanovirin-N with antiviral activity and preparation as well as modification of its polyclonal antibody].
abstract: OBJECTIVE
  To purify the recombinant Cyanovirin-N (CVN) and determine its anti-influenza virus A (H1N1) activity, and to prepare the polyclonal antibody of CVN, purify it and label it with an enzyme for future application.
  
  
  METHODS
  The recombinant CVN were rapidly purified by two rounds of Ni-NTA chelating chromatography intervened with a SUMO protease cleavage step. Anti-H1N1 activity of CVN was determined using cytopathogenic effect assay (CPE). The rabbits were immunized with the purified CVN and the antibody was identified by ELISA and Western blot. IgG was purified by ammonium sulfate precipitation followed by DEAE chromatography and the purified IgG was labeled by HRP.
  
  
  RESULTS
  The purity of the obtained CVN protein which showed obvious Anti-H1N1 activity in vitro was higher than 95%.The polyclonal antibody of CVN was successfully produced in the rabbits and the results of ELISA and Western blot showed that the antiserum had high titer and high specificity. The purified antibody with a titer up to 1:6 400 was successfully obtained and the anti-CVN antibody-HRP conjugate was achieved after labeling the purified antibody with HRP.
  
  
  CONCLUSIONS
  The purified antibody against CVN has been purified and further coniugated with HRP, with can be used for future research.
authors.affiliation: Biomedical Research and Development Center, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510630, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Han
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.forename: Qiao-li
authors.initials: QL
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Chui-wen
authors.initials: CW
authors.lastname: Qian
authors.forename: Sheng
authors.initials: S
authors.lastname: Xiong
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D000998:Antiviral Agents / Q000276:immunology / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification*; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D011817:Rabbits
chemical_list: D000906:Antibodies; D000998:Antiviral Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; C107733:cyanovirin N