production and purification of polyclonal antibodies against diphtheria toxin

diphtheria is a fatal disease caused by exotoxin of corynebacterium     diphtheria .  this toxin consists of   two chains, catalytic chain (a) and binding (b) chain. by binding chain (b),   the toxin binds to its receptor on numerous body cells such as myocardial,   kidney and peripheral nerve cells. after entering, catalytic chain (a)   inhibits protein synthesis and finally can cause cell death. at this time,   the toxoid form of diphtheria toxin is used as vaccine. the aim of this study   was the   immunological analysis of the mutated synthetic catalytic subunit of   diphtheria toxin in laboratory animals as a vaccine candidate, in addition to   polyclonal antibody   production and purification against diphtheria toxin. for this purpose the dtx     recombinant protein (with two mutant: a158g and g52e) was expressed using   pet28a/dtxa plasmid in e. coli bl21de3 host. then, recombinant   protein, as a   candidate vaccine, was extracted and purified. after evaluating and   confirming the protein by sds-page and western blotting, immunization carried   out in laboratory animals. finally, followed by antibody titration by elisa,   antibody purification performed as well.the mutated recombinant protein   prepared from an optimized expression was extracted and purified. then, this   protein was confirmed by sds-page and western blotting. elisa results showed   a satisfactory immunization of animals by this protein. polyclonal antibody   production and purification against diphtheria toxin was performed by g   protein column and confirmed by elisa.   elisa results showed a high titer of polyclonal antibody against diphtheria   toxin in animal's serum after immunization by recombinant dtx protein.