Overexpression, and Purification of Ovalbumin and its Variants

Abstract

The growing population in many regions of the world has raised concerns about the increased risk of infectious diseases. As more people inhabit the planet, there is a higher potential for the spread of pathogens and the emergence of new infectious diseases resulting in a higher disease burden. This has significant implications for global health and necessitates proactive measures to address these concerns. One promising strategy to combat infectious diseases is the use of vaccines, although not all vaccines possess the necessary factors to generate a strong immune response. Thus, various approaches have been developed to enhance immunogenicity such as adjuvants; platforms like nanoparticles, and VLPs; carriers like dendrimers, liposomes, and antigen scaffolds have shown effective results but are still challenged by various drawbacks like generating toxicity, allergy, difficulty to generate or manufacture, and high production cost. Hence, several strategies have been developed that will lead to multimerization or oligomerization of the antigen followed by increased immunogenicity. Protein multimers and oligomers have shown greater efficacy to induce an effective immune response against various diseases due to their larger size which can be easily recognized by the immune system. Thus, we propose that an amyloidogenic motif such as Lys-Phe-Phe-Glu (KFFE) when attached to a protein could multimerize protein antigen leading to enhanced immunogenicity and presenting antigen to antigen-presenting cells (APCs) easily and is a safer, simpler and cost-effective approach. During our project, we were able to clone and express the OVA (model antigen) and OVA-tag (OVA-KFFE) in E coli. BL21 strain, whereas purified protein is yet to be confirmed by western blot.