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      Expression, purification and evaluation of recombinant lipoprotein of Salmonella typhi as a vaccine candidate.

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          Abstract

          Lipoprotein has been reported as a vaccine candidate against many pathogenic bacteria, it plays direct role as a virulence-associated function. Here the approach is toward the expression of recombinant lipoprotein of Salmonella typhi in prokaryotic host and its evaluation as a vaccine candidate. Lipoprotein gene (lp1) was cloned in pET32a expression vector in addition to Bam HI and Hind III restriction sites, and BL21(pLysS) was used as prokaryotic expression host for transformation. Lipoprotein induction was performed by IPTG and 55 kDa (31 kDa of Gene +24 kDa of vector additional protein with His-tag) was analyzed by 12% SDS-PAGE. The recombinant lipoprotein was purified by Ni-NTA affinity chromatography due to the addition of 6X His-tag in recombinant lipoprotein. Western blot analysis using anti-His tag polyclonal antibodies confirmed the specificity of recombinant lipoprotein. Immunogenicity and protection study of recombinant lipoprotein against S. Typhi was performed in BALB/c mice. Adjuvants IFA and alum salts were used to enhance the immune response. ELISA results proved that biologically active truncated recombinant lipoprotein (31 kDa) is a suitable immunogen. Alum salts used as adjuvant was effective for long-lasting immunity.

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          Author and article information

          Journal
          Biologicals
          Biologicals : journal of the International Association of Biological Standardization
          Elsevier BV
          1095-8320
          1045-1056
          Mar 2017
          : 46
          Affiliations
          [1 ] School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi, India. Electronic address: Anand411@gmail.com.
          [2 ] School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi, India. Electronic address: skundu.bce@itbhu.ac.in.
          [3 ] School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi, India. Electronic address: m.debnath.bce@itbhu.ac.in.
          Article
          S1045-1056(17)30006-4
          10.1016/j.biologicals.2017.01.008
          28189484
          ca1063b7-ef64-428e-9f8d-97dfed7e5607
          History

          Salmonella typhi,Immune response,Incomplete Freund's adjuvant,Lipoprotein,Vaccine

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