Antibodies are polyclonal antibody production tools for a wide range of biomedical applications. Polyclonal antibody production involves immunizing animals, such as rabbits, mice, guinea pigs or rats with an antigen and collecting the resulting antibodies in their serum. The resulting antibodies are often used in Western blotting, ELISA and immunohistochemistry. Compared to monoclonal antibodies, which target a single epitope, polyclonal antibodies are typically made up of a mixture of antibodies that recognize different epitopes and are therefore more versatile.

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The use of animals for antibody production raises ethical concerns, including stress and discomfort associated with injections and the potential toxicity of Freund’s adjuvant. Moreover, the sensitivity of antibody-producing cells can limit the number of antibodies that can be produced, and batch-to-batch variability can impact experimental results. Furthermore, antibody purification can be challenging and can result in the presence of impurities that may interfere with experimentation.

In this study, we utilize a DNA-gold micronanoplexed approach to generate polyclonal antibodies against membrane proteins from the pathogens F. tularensis and ASFV. We optimized a simple immunogen formulation containing pCMVi-LSrCOMPTT, PAAG-immobilized BDDrFVIII and a peptide to increase the number of target-specific antibodies and their titers. The titers of the generated antibodies were measured in an ELISA assay and confirmed by a Western blot assay. The generated antibodies recognized a number of the membrane protein targets, with titers of 1:1,000 or higher in comparison to naive controls. These results suggest that a simple, genetic immunization procedure can yield robust antibodies against membrane proteins from the pathogens and can be used for vaccine development.