Recombinant chimeric proteins containing epitopes of acinetobacter baumannii proteins for use as biotechnological input

Description

[001] The present invention, internationally classified under patent classifications A61K 39 / 02 (bacterial antigens), C12N 15 / 09 (recombinant DNA technology), C12N 15 / 10 (processes for purification, isolation or purification of DNA or RNA), C12N 15 / 31 (genes encoding microbial proteins), C12N 15 / 66 (general methods for inserting a gene into a vector to form a recombinant vector using cleavage or ligation), C12N 15 / 70 (vectors or expression systems specially adapted to E. coli), and G01N 33 / 53 (immunoassay). The invention relates to the construction of a multi-epitope chimeric protein, from immunogenic epitopes of Acinetobacter baumannii proteins, designed using in silico analyses and immunoinformatics tools. The chimera is produced in a heterologous expression system and its use as a vaccine or immunotherapy in the treatment of infections associated with A. baumannii is proposed. Background of the Invention

[002] The genus Acinetobacter is composed of coccobacillary, gram-negative, aerobic, non-motile, catalase-positive and oxidase-negative bacteria, belonging to the family Moraxellaceae and the order Gammaproteobacteria (PELEG, AY; SEIFERT, H.; PATERSON, DL Acinetobacter baumannii: Emergence of a successful pathogen. Clinical microbiology reviews, v. 21, n. 3, p. 538-582, 2008.MARTINS, AF; BARTH, AL Multiresistant Acinetobacter - a challenge for the Petition 870250044128, dated 05 / 28 / 2025, p. 6 / 21 2 / 16 public health. Scientia medica, vol. 23, no. 1, p. 56, 2013). Widely found in soil and water, this bacterium can survive on inanimate and dry surfaces for several months and grow in high temperatures and pH variations (NOWAK, P.; PALUCHOWSKA, P. Acinetobacter baumannii: biology and drug resistance — role of carbapenemases. Folia histochemica et cytobiologica, 2015), with the species A. baumannii being the most relevant of the genus due to its high pathogenicity and ability to adapt to multiple environments. Acinetobacter baumannii is considered an opportunistic pathogen that mainly affects hospitalized and immunocompromised patients (HAWLEY, JS et al. Susceptibility of Acinetobacter strains isolated from deployed US military personnel). Antimicrobial agents and chemotherapy, v. 51, n. 1, p. 376378, 2007; SHEPPARD, F. R. et al. The majority of US combat casualty soft-tissue wounds are not infected or colonized upon arrival or during treatment at a continental US military medical facility. American journal of surgery, v. 200, n. 4, p. 489-495, 2010), é responsável por aproximadamente 12% das infecções hospitalares em todo o mundo (HUANG, Y. et al. Acinetobacter baumannii ventilator-associated pneumonia: Clinical efficacy of combined antimicrobial therapy and in vitro drug sensitivity test results. Frontiers in pharmacology, v. 10, 2019.; GELLINGS, P. S.; WILKINS, A. A.; MORICI, L. A. Recent Advances in the Pursuit of an Effective Acinetobacter baumannii Vaccine. Pathogens, v. 9, n. 12, p. 1066, 2020). Infections caused by this pathogen are associated with organs and systems with high rates of fluid exchange, such as the urinary tract, respiratory system, and peritoneal system (MORRIS, D. et al. Dissemination of clonally related multidrug-resistant Klebsiella pneumoniae in Ireland. Epidemiology and infection, v. 144, n. 2, p. 443-448, 2016.). Petition 870250044128, dated 05 / 28 / 2025, page 7 / 21 3 / 16

[003] The prevalence of A. baumannii as responsible for significant hospital outbreaks, especially in ICUs, has been made possible by the acquisition of antimicrobial resistance and the ability to survive in the hospital environment. Since the 1970s, there has been an increase in the number of cases of infections caused by this bacterium and in the occurrence of multidrug-resistant strains, leading to high mortality rates and a scarcity of therapeutic options. A. baumannii has shown significant resistance to several antimicrobial drugs (VILAS BOAS, DP A. Evaluation of the mechanisms of interaction between host and phage in biofilms. Escola de Engenharia Brasil, October, p. 121, 2013) and its virulence factors, including biofilm production and outer membrane proteins, have been associated with its persistence and ability to escape the host's immune response (ROCA, I. et al.The Acinetobacter baumannii oxymoron: commensal hospital dweller turned pan-drug-resistant menace, [n.d.]. Available at: https: / / diposit.ub.edU / dspace / bitstream / 2445 / 138641 / 1 / 62832 9.pdf), representing a significant risk to public health.

[004] Given the growing need to develop new therapeutic strategies against A. baumannii, this research presents an innovative approach, using immunoinformatics and in silico analyses as promising strategies to study A. baumannii and identify potential therapeutic targets. Previously, antibody design and vaccine development were expensive and time-consuming; however, with the advances in bioinformatics currently available, practical tools have been developed that can be used to reduce the time and cost of vaccine and antibody production (ANDERSON, LJ et al. Strategic priorities for respiratory syncytial virus (RSV) vaccine development. Vaccine, v. 31, p. B209-B215, 2013; KAZI, A. et Petition 870250044128, dated 05 / 28 / 2025, page 8 / 21 4 / 16 al. Current progress of immunoinformatics approach harnessed for cellular- and antibody-dependent vaccine design. Pathogens and global health, v. 112, n. 3, p. 123-131, 2018; OLI, A. N. et al. Immunoinformatics and vaccine development: An overview. ImmunoTargets and therapy, v. 9, p. 13-30, 2020.).

[005] A key strategy in this research is the engineering of chimeric proteins (CHEN, C. et al. Designing, expression and immunological characterization of a chimeric protein of Mycoplasma pneumoniae. Protein and peptide letters, v. 23, n. 7, p. 592-596, 2016), which incorporates multiple epitopes capable of stimulating robust and specific immune responses against the pathogen, thus increasing the potential of the therapeutic approach. According to the literature, vaccines produced from multiple epitopes have an advantage over classic vaccines due to their greater efficiency, lower chance of cross-reactivity, cost-effectiveness, and ability to induce targeted immune responses (SETTE, A. et al. The development of multi-epitope vaccines: Epitope identification, vaccine design and clinical evaluation. Biologicals: journal of the International Association of Biological Standardization, v. 29, n. 3-4, p. 271-276, 2001; GOUMARI, MM et al.Multi-Epitope Vaccines (MEVs), as a novel strategy against infectious diseases. Current proteomics, v. 17, n. 5, p. 354-364, 2020). In this context, using bioinformatics servers, epitopes can be predicted with high precision and accuracy (SORIA-GUERRA, RE et al. An overview of bioinformatics tools for epitope prediction: Implications on vaccine development. Journal of biomedical informatics, v. 53, p. 405-414, 2015).

[006] This technique allows for a prior understanding of the possible immune response generated by these constructs, enabling adjustments before protein synthesis. The construction of multi-epitope chimeras is a process that involves selection Petition 870250044128, dated 05 / 28 / 2025, page 9 / 21 5 / 16 of target antigens, epitope prediction, and evaluation of the immunogenic, allergenic, toxicological, and physicochemical properties of the designed proteins, and, in this research, based on a literature review, the choice of antigens was based on their clinical and pathological relevance in the host adhesion process and infection by A. baumannii, using databases such as UniProt, an essential tool in scientific research that allows the functional annotation of new sequences, the study of protein evolution, and the identification of potential therapeutic targets (THE UNIPROT CONSORTIUM et al. UniProt: the universal protein knowledgebase in 2021. Nucleic acids research, v. 49, n. D1, p. D480-D489, 2021).

[007] From the antigens selected for this study, eight epitopes were used to design two chimeric proteins, which were then synthesized and cloned into appropriate expression vectors for recombinant production. The efficacy of these multi-epitope chimeras depends on their ability to induce an immune response, their safety, and the absence of adverse effects. Thus, the in silico analysis of the antigenicity, allergenicity, and toxicity of the chimeras was crucial, as it reduces costs and time compared to laboratory tests (AYYAGARI, VS. Design of linear B cell epitopes and evaluation of their antigenicity, allergenicity, and toxicity: An immunoinformatics approach. In: Computational Vaccine). Design. New York, NY: Springer US, 2023. p. 197-209). In this sense, this study aims to develop, produce, and characterize two chimeric proteins containing immunogenic epitopes of the bacterium A. baumannii, expecting that the use of immunoinformatics, in silico analysis, and molecular biology approaches in this research will contribute to the development of new therapeutic strategies against A. baumannii infections. Petition 870250044128, dated 05 / 28 / 2025, page 10 / 21 6 / 16 Description of the Invention

[008] The target antigens for the construction of A. baumannii multiepitope chimeras were chosen based on a literature review. Proteins of clinical and pathological relevance in host adhesion and the A. baumannii infection process were selected. Table 1 lists the eight antigens chosen. Protein sequences for each antigen were obtained from the UniProt database (https: / / www.uniprot.org / ). All sequences were retrieved in FASTA format for subsequent bioinformatic analyses.

[009] Potential epitopes capable of stimulating lymphocyte-mediated immune responses were predicted in the protein sequences of selected antigens using the following servers: NetMHCpan v. 4.1 (REYNISSON, B. et al. NetMHCpan-4.1 and NetMHCIIpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data. Nucleic acids research, v. 48, n. W1, p. W449-W454, 2020.), NetMHCIIpan v. 4.0 (REYNISSON, B. et al. NetMHCpan-4.1 and NetMHCIIpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data. Nucleic acids research, v. 48, n. W1, p. W449-W454, 2020.) and BepiPred v. .0 (CLIFFORD, JN et al. BepiPred-3.0: Improved B-cell epitope prediction using protein language models. Protein science: a publication of the Protein Society, v. 31, n. 12, 2022). The NetMHCpan and NetMHCIIpan servers were used to predict epitopes recognized by lymphocytes. Cytotoxic T lymphocytes (CTLs) and helper T lymphocytes (HTLs), respectively. Petition 870250044128, dated 05 / 28 / 2025, page 11 / 21 7 / 16 Table 1: Selection of antigen epitopes Antigen Function Ata Adhesion to host cells and biofilm formation Bap Biofilm formation and adhesion to surfaces BamA Assembly of outer membrane proteins BauA Mediation of iron acquisition FhaC facilitates the secretion of a hemagglutinin protein. Selected filamentous cells and their respective functions. Epitopes MNQIADTNKNLAKKVDQLTKVKDNLRITDTNNQLN ADLIGAIDGGTDGTFNAQVAL MLEDLPVMPVDQNMANEIYRVAEEAKVRYRLDRLTQKDISELPDA FKTAASEQEALLF VDVGRRFGENKEFGVRINGMYRDGDAAVNDSRLFSLGLDWQGNAR VFVDAYDALNHTQDDYGYRIIPGFSDPVITNIYDPNPNWTASGEI FPPQKTKQQELGLKVDLGTFAHTLSAFEITKPSSYLDPSKLVNNP TFVSDGEQRNRGIEWSFFGSPIEHVRLMGGFTYLDP VSLPSQVLQDQRLKELNQQLQDQLAQKVNFNLSVSGNQDTGTYMN IGIGINNPIHLNDILSLNVSHSLDNFHEDLNRSYFISYQLPVGYY DLGFSYNDYQYKQTVLGGNGPLINVLHRRTSGWNLGFQHRQYLGN AVLDGSIDYRRGMGVGARTAPEENITDVYGNHLPVEGYSRAP Petition 870250044128, dated 05 / 28 / 2025, page 12 / 21 8 / 16 OmpA plays a role in bacterial adhesion and virulence. Omp33-36 functions as an important outer membrane protein, potentially involved in antibiotic resistance. Oxa-143 confers resistance to beta-lactam antibiotics. WLGFEAEYDGASAGAEYKQKQINGNFYVTSDLITKNYDSKIKPYV LGAGHYKYDFDGPVETPVAPQPQELTEDLNMELRVFFDTNKSNIK DQYKPEIAKVAEKLSEYPNATARIEHTDNTGPKL TYGVKGEAYVPTPYLPVYASATYNHSKDDNGDRYALEVGAMLLPL MTVGYTSVANQFALDNFGIIGNGIYSAVNQTAAIQNDQDAVTARA KYVGPIDGTNMAIGFEAAGAFGQENQYGLKTYLTPKLSADIKGND LGEFRQAWGGNVNYFITP VYQELARRTGLDLMQKEVKRVGFGNMNIGTQVDNFWLVGPLKITP IQEVNFADDFANNRPFKLETQEEVKKMLLIKEFNGSKIY Petition 870250044128, dated 05 / 28 / 2025, page 13 / 21 9 / 16

[009] Both servers use a pan-MHC approach capable of predicting protein binding to multiple alleles. MHC class I and II. BepiPred was used to predict epitopes capable of being recognized by B cells. Predefined parameters were used for all analyses. Potential epitopes capable of stimulating lymphocyte-mediated immune responses were predicted in the protein sequences of the selected antigens using the following servers: NetMHCpan v. 4.1 (REYNISSON, B. et al. NetMHCpan-4.1 and NetMHCIIpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data. Nucleic acids research, v. 48, n. W1, p. W449-W454, 2020.). NetMHCIIpan v. 4.0 (REYNISSON, B. et al. NetMHCpan-4.1 and NetMHCIIpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data. Nucleic acids research, v. 48, n. W1, p. W449-W454, 2020.) and BepiPred v. 3.0 (CLIFFORD, JN et al. BepiPred-3.0: Improved B-cell epitope prediction using protein language models. Protein science: a publication of the Protein Society, v. 31, n. 12, 2022.). The NetMHCpan and NetMHCIIpan servers were used to predict epitopes recognized by cytotoxic T lymphocytes (CTLs) and helper T lymphocytes (HTLs), respectively. Both servers use a pan-MHC approach capable of predicting the binding of proteins to multiple MHC class I and II alleles. BepiPred was used to predict epitopes capable of being recognized by B cells. Predefined parameters were used for all analyses.

[010] To develop a multi-epitope construct, all selected epitopes were linked via polyglycine ligands (GGGGG). These ligands were chosen by Petition 870250044128, dated 05 / 28 / 2025, page 14 / 21 10 / 16 its flexibility and stability, conferring protein-protein binding (REDDY CHICHILI, VP; KUMAR, V.; SIVARAMAN, J. Linkers in the structural biology of protein-protein interactions. Protein science: a publication of the Protein Society, v. 22, n. 2, p. 153-167, 2013.). Two distinct multiepitope chimeras were constructed, one with a final sequence composed of 424 residual amino acids and the other with 448 residual amino acids. The distribution of epitopes and linkers in the chimeras is illustrated in Figure 1.

[011] The VaxiJen v. 2.0 server (DOYTCHINOVA, IA; FLOWER, DR VaxiJen: a server for prediction of protective antigens, tumor antigens and vaccine subunits. BMC bioinformatics, v. 8, n. 1, 2007) was used to predict the antigenicity of the protein sequences. Bacteria were selected as the target organism and a threshold of 0.5 was established. The non-allergenic behavior of the chimeras was predicted using AllerTOP v. 2.0 (DIMITROV, I. ​​et al. AllerTOP v.2—a server for in silico prediction of allergens. Journal of molecular modeling, v. 20, n. 6, 2014.), which predicts the allergenic potential of protein sequences based on a cross-autocovariance transformation of the protein sequences into uniform vectors of equal length, followed by classification by a k-nearest neighbor algorithm. The ToxinPred2 server (SHARMA, N. et al. ToxinPred2: an improved method for predicting protein toxicity. Briefings in bioinformatics, v. 23, n. 5, 2022.) was used to predict the toxicity potential of the sequences. The results of these analyses can be seen in Table 2. Petition 870250044128, dated 05 / 28 / 2025, page 15 / 21 11 / 16 Table 2. Evaluation of antigenicity, allergenicity, and toxicity. Chimera 1 Chimera 2 Antigenicity Antigenic Antigenic (0.7196) (0.7074) Allergenicity Non-allergenic Non-allergenic Toxicity Non-toxic Non-toxic

[012] The physicochemical properties of the proteins were analyzed using the ProtParam tool. The characteristics evaluated included an overall mean hydropathy (GRAVY), instability index (II), half-life (HL), isoelectric point (pI), and molecular weight (MW). In addition, the solubility of the proteins was predicted using the SOLpro program. The results of these analyses can be seen in Table 3. Petition 870250044128, dated 05 / 28 / 2025, page 16 / 21 12 / 16 Table 3. Analysis of physicochemical properties Properties Chimera 1 Chimera 2 Number of amino acids 424 442 Molecular weight 46477.42 48684.67 Isoelectric point (pI) 5.12 4.86 Instability index Stable Stable Half-life (mammalian) 30 hours 30 hours GRAVY -0.508 -0.432 Aliphatic index 79.29 76.83 Solubility Soluble Insoluble

[013] The previously identified epitope sequence was used to generate a chimeric design in silico. This sequence was sent to GenOne (Rio de Janeiro, Brazil) for chemical synthesis, and the gene was cloned into the pet expression vector. The vector containing the recombinant sequence was heat-transformed into competent One Shot™ BL21 Star™ (DE3) cells and cultured in 250 mL of broth. 2.5 mL of 2m glucose and kanamycin (SigmaAldrich, USA) at a concentration of 1 µg / mL were incubated at 37°C and 150 rpm until an OD of 0.6–0.8 was achieved. Expression was then induced with 250 µg of 1 mM isopropyl-β-1-D-thiogalactopyranoside (1 mM IPTG) and maintained under similar incubation conditions for a further 4 hours, then centrifuged (10,000 x g, 4°C, 15 min). Following this, a cell washing step was performed, in which the cells were resuspended in 15 mL of washing buffer (30 mM KCl, 15 mM KH2PO4, 1.4 M NaCl, 80 mM Na2HPO4, 0.1% Petition 870250044128, dated 05 / 28 / 2025, pp. 17 / 21 13 / 16 The pellets were then centrifuged again using Triton X100, pH 7.4. The resulting pellet was stored at -20°C for 12 hours and subsequently resuspended in 15 mL of solubilizing buffer (20 mM NaH2PO4, 0.5 M NaCl, imidazole 40, pH 7.4) plus 25 µL of lysozyme and 25 µL of phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich, USA). The suspension was shaken at 37°C for 1 hour and then placed in an ice bath for 15 minutes. Subsequently, the cells were lysed using a sonicator (Ultrasonic Processor; GE Healthcare Bio-Sciences Corp. Piscataway, NJ, USA) and centrifuged (10000 x g, 4°C, 40 min) to obtain the soluble portion. To obtain the insoluble portion, the pellet obtained from the soluble portion was suspended in a solubilizing buffer plus urea (8M urea, 20 mM NaH2PO4, 0.5M NaCl, imidazole 40, pH 7.4), which was kept under agitation at 4 °C for 16 hours and then centrifuged again.

[014] Both portions were purified by affinity chromatography using nickel-loaded HisTrap FF columns (GE Healthcare) in the Akta Purifier automated liquid chromatography system (GE Healthcare, Chicago, IL). To complete the production process and reduce the urea concentration in the insoluble portion, the proteins were dialyzed against 1X phosphate-buffered saline (PBS) in 7 steps at 4°C for two days. Subsequently, to confirm the expression and purity of the chimera, 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), DOT Blotting (DB), and Western Blotting (WB) were performed. In addition, the protein concentration was determined using the BCA Protein Assay kit (Pierce, USA), and both portions were stored at -20°C.

[015] Western blotting was performed against anti-6*Histag. An aliquot of the purified chimera was loaded onto a gel. Petition 870250044128, dated 05 / 28 / 2025, pages 18 / 21 14 / 16 A 12% SDS-PAGE sample was separated and transferred to a hybond™ ECL™ nitrocellulose membrane (Amersham Biosciences) using the Trans-Blot Turbo™ transfer system. A PBS solution with 5% milk powder was then used to block nonspecific sites on the membrane for one hour. Subsequently, the membrane was incubated overnight with anti-6*Histag monoclonal antibody (SigmaAldrich, USA) at a 1:50 dilution under 50 rpm agitation. After a wash step with 1X PBS to visualize the protein bands, the reaction was carried out with a chromogen / substrate solution (0.012 g diaminobenzidine, 0.03% nickel sulfate, 50 mM Tris-HCl, pH 7.6, and 0.03% hydrogen peroxide). The result can be seen in Figure 2.

[016] DOT blotting was performed against anti-6*Histag, mouse serum, and human serum. A 5μL aliquot of the purified chimera was loaded onto hybond™ ECL™ nitrocellulose membranes (Amersham Biosciences) and allowed to dry for 15 minutes at 37°C. A PBS solution with 5% milk powder was then used to block nonspecific sites on the membranes for one hour; subsequently, the membranes were individually incubated with primary mouse serum (Sigma-Aldrich, USA), human serum (Sigma-Aldrich, USA), and anti-6*Histag monoclonal antibody (Sigma-Aldrich, USA) at a 1:50 dilution overnight under agitation at 50 rpm. After a washing step with 1X PBS, membranes containing human and mouse sera were incubated overnight with peroxidase-conjugated human anti-IgG secondary antibody (Sigma-Aldrich, USA) and mouse secondary antibody (Sigma-Aldrich, USA), both at a 1:5000 dilution in PBS.However, the anti-6*Histag monoclonal antibody membrane was kept in solution overnight. To visualize the protein bands, a. Petition 870250044128, dated 05 / 28 / 2025, pp. 19 / 21 The reaction 15 / 16 was developed with a chromogen / substrate solution (0.012 g diaminobenzidine, 0.03% nickel sulfate, 50 mM Tris-HCl pH 7.6 and 0.03% hydrogen peroxide). The result can be seen in Figure 3. Brief Description of the Tables and Figures Table 1 illustrates the final selection of epitopes from the selected antigens, their respective functions, and sequences. Table 2 presents the results of the antigenicity, allergenicity, and toxicity assessments performed on chimeras 1 and 2. Table 3 shows the results of the analyses of the physicochemical properties applied to both chimeras. The results include molecular weight, number of amino acids, isoelectric point, solubility, half-life, GRAVY spectroscopy, stability index, and aliphatic index. Figure 1 presents a graphical representation of the structures of chimeras 1 (A) and 2 (B). Figure 2 shows the result of chimera expression by Western blotting against anti-6x-histidine monoclonal antibody, being: (M) PageRuler Prestained Protein Ladder (Thermo Scientific), (1) Chimera 1 and (2) Chimera 2. Figure 3 shows the antibody recognition capacity of chimeric proteins through the DOT Blotting assay. A: DB with anti-6xhistidine monoclonal antibody, (C+) rCAM87009.1 protein, (C-) Negative control, (1) Chimera 1 and (2) Chimera 2; B: BD with serum from a human infected with A. baumannii (1:100 dilution), (C+) rCAM87009.1 protein, (C-) Negative control, (1) Chimera 1 and (2) Chimera 2; C) DB with mouse serum (1:100 dilution). Petition 870250044128, dated 05 / 28 / 2025, pages 20 / 21 16 / 16 1:50), (C+) rCAM87009.1 protein, (C-) Negative control, (1) Chimera 1 and (2) Chimera 2.

Claims

1) Recombinant chimeric proteins containing Acinetobacter baumannii protein epitopes for use as a biotechnological input, characterized by nucleotide sequences (SEQ ID No. 1) that encode immunogenic epitopes of the Ata, BauA, Bap and FhaC proteins of Acinetobacter baumannii; 2) Recombinant chimeric proteins containing Acinetobacter baumannii protein epitopes for use as a biotechnological input, characterized by a nucleotide sequence (SEQ ID No. 2) encoding antigenic epitopes of the BamA, Oxa-143, OmpA and Omp33-36 proteins of A. baumannii; 3) Recombinant chimeric proteins containing Acinetobacter baumannii protein epitopes for use as a biotechnological input, as per claims 1 and 2, characterized by being used for cloning genes and expressing chimeric proteins in prokaryotes, eukaryotes or mammals, but not limited to these; 4) Recombinant chimeric proteins containing Acinetobacter baumannii protein epitopes for use as a biotechnological input, as claimed in claims 1, 2 and 3, characterized by using a recombinant expression vector in Escherichia coli, followed or not by purification of the chimeric protein; 5) RECOMBINANT CHIMERIC PROTEINS CONTAINING EPITOPES OF Acinetobacter baumannii PROTEINS FOR USE AS A BIOTECHNOLOGICAL INPUT, as claimed in claims 1, 2, 3 and 4, characterized by composing individual or combined vaccine formulations to induce a specific immune response against A. baumannii; Petition 870250044128, dated 05 / 28 / 2025, page 4 / 21 2 / 2 6) RECOMBINANT CHIMERIC PROTEIN CONTAINING EPITOPES OF Acinetobacter baumannii PROTEINS AND ITS USE AS A BIOTECHNOLOGICAL INPUT, as per claims 1, 2, 3, 4 and 5, characterized by being used in the production of antibodies for use in immunotherapies.

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