title: [Preparation of monoclonal antibody against Pfs25 protein and establishment of double antibody sandwich ELISA method].
abstract: OBJECTIVE
  To prepare the monoclonal antibody (mAb)against Pfs25 protein of Plasmodium falciparum, and establish the method of sandwich ELISA for detecting the Pfs25  protein.
  
  
  METHODS
  Pfs25 protein the recombinant expressed by Pichia pastoris was purified. The purified Pfs25 protein as the antigen was used to immune the BALB/c   mice,  The secreting specific mAb positive cell strains, which were prepared by hybridizing the Sp2/0 myeloma cell and the spleen cell from immunized mice, were detected by indirect ELISA method. The ascites of mAb were collected from immunization F1 mice, and their biological properties were identified by indirect ELISA. The anti-Pfs25 antibody was labeled by Horseradish Peroxidase (HRP),  the sandwich ELISA method to detect Pfs25 protein was established based on the anti-Pfs25 mAb  4B7 and 1B4 as coating and enzyme antibody,  respectively.
  
  
  RESULTS
  Three hybridoma cell lines secreting mAb against Pfs25 protein have been selected from the antibody positive hybridizing cells. The two of them have a better stability and specificity. The sandwich ELISA method detecting Pfs25 protein was established. Its detecte range was 0.07-1  mg/mL , and its sensitivity was 41.6 ng/mL.
  
  
  CONCLUSIONS
  The anti-Pfs25 mAb are  successfully prepared and the double antibody sandwich ELISA method detecting Pfs25 protein  is established. Our study lay a foundation of developing transmission-blocking malaria vaccine with Pfs25 protein as antigen.
authors.affiliation: Lanzhou Institute of Biological Products, Lanzhou 730046, China. lilac_bio@yahoo.com.cn
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Ya-ru
authors.initials: YR
authors.lastname: Ma
authors.forename: Jian
authors.initials: J
authors.lastname: Lu
authors.forename: Qing
authors.initials: Q
authors.lastname: Lei
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Lin
authors.initials: L
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D015800:Protozoan Proteins / Q000032:analysis / Q000276:immunology*; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; C071284:Pfs25 protein, Plasmodium falciparum; D015800:Protozoan Proteins