Browsing by Author "Orellana Benenaula, Juan Diego"

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    Generación del antígeno IalB de Bartonella henselae y una versión modificada por la adición de un péptido con afinidad al poliestireno
    (Universidad de Cuenca, 2024-08-05) Encalada Ochoa, Edgar Vinicio; Orellana Benenaula, Juan Diego; Vallecillo Maza, Antonio Javier
    Bartonellosis is a chronic and inapparent disease caused by Gram-negative bacteria of the Bartonella genus. Bartonella species are characterized by affecting multiple mammals, such as felines, canines, ruminants, rodents, humans, among other hosts. Domestic felines (Felis catus domesticus) act as reservoirs for Bartonella henselae; they can transmit the infection through different blood-sucking vectors, such as fleas, ticks, lice, mites, among others. Due to the absence of a sufficiently reliable indirect diagnostic test and the high frequency of infection reported in felines, the generation of the invasion-associated locus protein B (IalB), a protein located in the outer membrane of Bartonella henselae, was proposed and It is involved in intraerythrocytic adhesion and invasion. To obtain the coding sequence of IalB, assembly was carried out by the polymerase chain reaction (PCR)-thermodynamically balanced inside-out (TBIO) method. For the expression of IalB and an IalB-PoBP variant (Polystyrene binding peptide (PoBP) added to the carboxyl end), the Escherichia coli Rosetta 2 (DE3) strain was used; both antigens were purified by immobilized metal affinity chromatography (IMAC), having a yield of approximately 4,50 mg of pure protein per liter for IalB and 1,39 mg of protein for the case of IalB-PoBP. With both antigenic proteins obtained, an indirect ELISA test was carried out in order to evaluate the recognition by antibodies of domestic felines, obtaining similar results for both antigens, which should be evaluated with sera from animals previously identified as infected by Bartonella henselae