Production and characterization of novel monoclonal antibodies against outer membrane protein A (OmpA) of Acinetobacter baumannii

Abstract

Background: Infection caused by Acinetobacter baumannii has emerged as a significant clinical problem with unacceptably high mortality rate due to the increase in antibiotic-resistant strains. Producing novel monoclonal antibody (MAb) against outer membrane protein A (OmpA) could be considered as a potential tool to improve treatment of A. baumannii infections.

Objectives: In this study, we aimed to produce murine MAbs against OmpA peptide of A. baumannii.

Materials and methods: BALB/c mice were immunized with 18-mer amino acid peptide as a part of the OmpA protein. Four antibody-secreting hybridomas were obtained using hybridoma technology and then characterized according to isotypes, affinity constant, reactivity in ELISA, flow cytometry, indirect immunofluorescence (IFA) and opsonophagocytic killing assays.

Results: All four produced MAbs (1A1-D10, 1G1-E7, 2C11-F10, and 4H2-H9) had IgG1 isotype with Kappa light chain. One of these MAbs, 1G1-E7 was purified and selected for further characterizations. 1G1-E7 showed a high reactivity with both immunogenic peptide and A. baumannii in ELISA. Our results indicated that 1G1-E7 MAb reacted with 95.3% of A. baumannii in flow cytometry as well as IFA. Moreover, the affinity of the 1G1-E7 MAb was measured 1.37 × 10⁸ M^-1. The 1G1-E7 significantly improved opsonophagocytic killing of a clinical isolate of A. baumannii.

Conclusion: Our findings showed that the OmpA can be identified by produced MAbs. The efficacy of novel anti-OmpA antibodies in A. baumannii targeting needs to be further investigated in challenging models, and then could be subjected for genetic engineering to produce therapeutic antibody against A. baumannii.

Keywords: Acinetobacter baumannii; ELISA; Flow cytometry; Indirect immunofluorescence assay (IFA); Monoclonal antibody (MAb); Outer membrane protein a (OmpA).