Record number :
2427198
Title of article :
Production and purification of polyclonal antibodies against Diphtheria toxin
Author/Authors :
Arefpour Torabi, Mohammad Ali Biology Research Center - Faculty and Institute of Basic Science - Imam Hossein University, Tehran, Iran , Olad, Gholam Reza Applied Biotechnology Research Center - Baqiyatallah University of Medical Sciences, Tehran, Iran , Nazarian, Shahram Biology Research Center - Faculty and Institute of Basic Science - Imam Hossein University, Tehran, Iran , Salimian, Jafar Chemical Injury Research Center - Baqiyatallah University of Medical Sciences, Tehran, Iran , Khodi, Samaneh Applied Biotechnology Research Center - Baqiyatallah University of Medical Sciences, Tehran, Iran , Bagheripour, Mohamad Javad Biology Research Center - Faculty and Institute of Basic Science - Imam Hossein University, Tehran, Iran
Pages :
5
From page :
67
To page :
71
Abstract :
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 SDSPAGE 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.
Keywords :
Diphtheria , Mutated DTxA chain , Diphtheria toxin , recombinant vaccine , Polyclonal antibody
Journal title :
Astroparticle Physics
Serial Year :
2014
Link To Document :
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