Cloning, Expression and Seroreactivity of the Recombinant Lipopolysaccharide Assembly Protein - D (LptD) from Bartonella Bacilliformis

Overview

Journal Rev Peru Med Exp Salud Publica

Specialties General Medicine
Public Health

Date 2022 Jun 29

PMID 35766735

Authors

Astrid Flores-Nunez

Gladis Ventura

Henri Bailon

Adolfo Marcelo

Gustavo Sandoval

Carlos Padilla-Rojas

Affiliations

Soon will be listed here.

Abstract

Objective.: To evaluate in silico and at the serological level the antigenic potential of the recombinant extracellular domain of the lipopolysaccharide assembly protein - D (LptD) of Bartonella bacilliformis (dexr_LptD).

Materials And Methods.: Through in silico analysis, we selected a B. bacilliformis protein with antigenic and immunogenic potential. 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 conducted.

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 carried out under denaturating conditions on a small scale and we obtained 2.6 μg/mL of partially purified dexr_LptD. 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.

Citing Articles

Cloning, expression and seroreactivity of the recombinant lipopolysaccharide assembly protein - D (LptD) from Bartonella bacilliformis.

Flores-Nunez A, Ventura G, Bailon H, Marcelo A, Sandoval G, Padilla-Rojas C Rev Peru Med Exp Salud Publica. 2022; 39(1):15-23.

PMID: 35766735 PMC: 11397601. DOI: 10.17843/rpmesp.2022.391.9292.

Carrion's Disease: the Sound of Silence.

Gomes C, Ruiz J Clin Microbiol Rev. 2017; 31(1).

PMID: 29187394 PMC: 5740975. DOI: 10.1128/CMR.00056-17.

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