Introduction
Our project aims to design a novel production method for antimicrobial peptides (AMPs) to replace the inefficient existing approaches. By sequestering AMPs within an encapsulin nanocompartment during expression, these peptides can be produced with reduced toxicity to their host, improving system productivity [1].
Additionally, encapsulin-facilitated AMP expression represents an advantage over chemical synthesis of AMPs for basic research, as it only requires basic molecular biology equipment instead of the various chemicals and techniques required for the lengthy process of chemical protein synthesis.
This manufacturing advancement would enable AMPs to be produced on a large scale and become a more realistic option for clinical antibiotic use. A conceptual lab-scale bioreactor design is shown below.
Figure 1: AMP + encapsulin bioreactor system
Customers
Our novel production strategy is intended to enable large-scale manufacturing of AMPs. We propose that the end users of this technology would include companies currently producing and consuming large quantities of antibiotics, such as pharmaceutical, chemical, and agricultural businesses.
Additionally, we propose creating an inexpensive and standardized cloning protocol to insert an AMP into a plasmid containing our TEV sites, linkers, targeting peptide, and encapsulin for easy use by other researchers. This could allow for accessible cloning, expression, and purification of AMPs to further exploratory research.
Regulatory
All drug manufacturing facilities in the United States must be registered with the Food and Drug Administration, so regulatory requirements must be considered. Because of the novelty of our synthetic biology-based production system, we expect the FDA approval and registration process to be one of the primary hurdles in the implementation process. The FDA has previously approved several AMPs for clinical therapeutic use, including Bacitracin, Telaprevir, and others [2]. However, sequestration of AMPs within encapsulins during bacterial expression is a novel concept, and has consequently not been evaluated by the FDA. Fortunately, the FDA does not require approval of new manufacturing processes but simply requires drug production to remain in line with Current Good Manufacturing Practice (CGMP) regulations. These are intended to ensure the final concentration and ingredients of the final drug product match what is claimed by the company. Therefore, additional quality control checks and safety protocols would need to be created in order to ensure production replicability and product purity.
References
- Lee, T-H, Carpenter, TS, D'haeseleer, P, Savage, DF, Yung, MC. Encapsulin carrier proteins for enhanced expression of antimicrobial peptides. Biotechnology and Bioengineering. 2020; 117: 603-613. https://doi.org/10.1002/bit.27222
- 2. Lei, J., Sun, L., Huang, S., Zhu, C., Li, P., He, J., Mackey, V., Coy, D. H., & He, Q. (2019). The antimicrobial peptides and their potential clinical applications. American journal of translational research, 11(7), 3919-3931.