Fusion expression and immunogenicity of Bordetella pertussis PTS1-FHA protein: implications for the vaccine development.
Mutants of pertussis toxin (PT) S1 subunit and filamentous hemagglutinin (FHA) type I immunodominant domain from Bordetella pertussis (B. pertussis) are considered to be effective candidate antigens for acellular pertussis vaccines; however, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. In this paper, the gene sequences of a S1 mutant C180-R9K/E129G (mS1) and a truncated peptide named Fs from FHA type I immunodominant domain were linked together and constructed to pET22b expression vector as a fusion gene; after inducing with IPTG, it was highly expressed in E. coli BL21 (DE3) as inclusion body. The fusion protein FsmS1 was purified from cell lysates and refolded successfully. The result of Western blotting indicate that it was able to react with both anti-S1 and anti-FHA McAbs; antiserum produced from New Zealand white rabbits immunized with this protein was able to recognize both native PT and FHA antigens as determined by western blotting. These data have provided a novel feasible method to produce PT S1 subunit and FHA type I immunodominant domain in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against B. pertussis infection.
Jinyong
Zhang
Z
Department of Clinical Microbiology and Immunology, The Third Military Medical University, Chongqing Engineering Technology Research Center of Biopharmaceuticals, Chongqing 400038, China.
Xiaoli
Zhang
Z
Weijun
Zhang
Z
Ying
Guo
G
Gang
Guo
G
Xuhu
Mao
M
Quanming
Zou
Z
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology; D015153:Blotting, Western; D001886:Bordetella pertussis / Q000276:immunology*; D003001:Cloning, Molecular; D007964:Leukocytosis / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D051379:Mice; D008807:Mice, Inbred BALB C; D037342:Pertussis Toxin / Q000276:immunology*; D010567:Pertussis Vaccine / Q000276:immunology*; D058849:Protein Refolding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification; D010566:Virulence Factors, Bordetella / Q000737:chemistry / Q000276:immunology*
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D010567:Pertussis Vaccine; D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; C083388:pertussis toxin, S1 subunit; D037342:Pertussis Toxin