title: [Optimization of bacterial display technology based on antigen-antibody co-expression].
abstract: OBJECTIVE
  To co-express antigen and antibody in E.coli periplasm in which the antigen and antibody will fold correctly and bind to each other.
  
  
  METHODS
  Human IL-1β gene and mouse anti-human IL-1β single chain variable fragment (anti-hIL-1β scFv) were inserted into the down stream from the NlpA and pelB leading peptides of pBFD vector respectively. The recombinant plasmid was named pBFD-Ab-Ag. The E.coli DH5α transformed with pBFD-Ab-Ag was induced by IPTG to co-express antigen and antibody. After the outer membrane of E.coli DH5α was broken (spheroplast formation), the bacteria were incubated with mouse anti-hIL-1β antibody labeled by FITC. The co-expression and interaction of antigen and antibody was analyzed by flow cytometry (FCM) based on fluorescence signal intensity.
  
  
  RESULTS
  The FCM detection showed the strong signal in E.coli DH5α transformed with pBFD-Ab-Ag and the positive rate was significantly improved.
  
  
  CONCLUSIONS
  The results showed that the antigen and antibody can fold and bind to each other efficiently in E.coli periplasm. The system can make antibody screening process easier and also provide a novel method for protein-protein interaction.
authors: Heilongjiang River Fishery Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China. lmxu0917@163.com
authors: Liming
authors: L
authors: Xu
authors: Mo
authors: M
authors: Guo
authors: Guiping
authors: G
authors: Ren
authors: Jiechao
authors: J
authors: Yin
authors: Deshan
authors: D
authors: Li
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000378:metabolism*; D000917:Antibody Formation; D000941:Antigens / Q000096:biosynthesis* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000906:Antibodies; D000941:Antigens; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D053583:Interleukin-1beta; D011993:Recombinant Fusion Proteins