Objective. To evaluate in silico and at the serological level the antigenic potential of the recombinant extracellular do-main of the lipopolysaccharide assembly protein-D (LptD) of Bartonella bacilliformis, hereinafter abbreviated as dexr_ LptD. Materials and Methods. Through in silico analysis, a B. bacilliformis protein with antigenic and immunogenic potential was selected. The selected protein gene was cloned into Escherichia coli TOP10 and expressed in Escherichia coli BL21 (DE3) pLysS. Recombinant protein was expressed using isopropyl-β-D-1-thiogalactopyranoside (IPTG) and induction conditions were optimized. Finally, it was purified with Ni-IDA resin (His60 Ni Superflow) and a Western Blot assay was performed. Results. In silico, the selected protein was LptD because it is located in the outer membrane and is antigenic and immunogenic. Optimized conditions for dexr_LptD induction were 0.5 mM IPTG, 16 hours, TB (Terrific Broth) medium, 3% (v/v) ethanol, 28 ºC, OD600: 1-1.5 and 200 rpm. Purification was performed under denaturating conditions on a small scale and 2.6 µg/mL of partially purified dexr_LptD was obtained. The Western Blot assay showed a positive reaction between the sera from patients with Carrión's Disease and dexr_LptD, which shows the antigenicity of dexr_LptD. Conclusions. The dexr_LptD shows antigenicity both in silico and at the serological level, these results are the basis for further studies on vaccine candidates against Carrion's Disease.
|Título traducido de la contribución||CLONING, EXPRESSION AND SEROREACTIVITY OF THE RECOMBINANT LIPOPOLYSACCHARIDE ASSEMBLY PROTEIN – D (LptD) FROM Bartonella bacilliformis|
|Número de páginas||9|
|Publicación||Revista Peruana de Medicina Experimental y Salud Publica|
|Estado||Publicada - 2022|
Nota bibliográficaPublisher Copyright:
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- Bartonella bacilliformis
- Bartonella infections
- Computational biology (source: MeSH NLM)
- LptD protein
- Recombinant proteins
- Vaccine Antigenicity