The COVID-19 Pandemic has revolutionized the field of mRNA vaccine technol., highlighting its utility and rapidly expanding potential, propelling it to the forefront of future vaccines & therapeutics for cryptic diseases.T7 RNA Polymerase (T7 RNAP) is a high-fidelity enzyme used for the synthesis of single-stranded mRNA from DNA templates using In vitro transcription (IVT).T7 RNAP is now the mainstay for the development of mRNA vaccines and therapeutics, accounting for 34% of the overall raw material cost for mRNA manufacturingAlthough small-scale purification processes for T7 RNAP exist, scalable process platforms and comprehensive anal. characterization are still lacking.Moreover, to address global raw material supply chain issues and to facilitate the efficient and large-scale production of mRNA vaccines & therapeutics, here we report a three-step purification process platform, yielding ∼85-100 mg/L of T7 RNAP from E. coli fermentation harvest.Also, to demonstrate its high-level purity and structural integrity, we have characterized it using orthogonal anal. techniques.Furthermore, when evaluated in an IVT system, this highly pure T7 RNAP showed activity that was comparable to the com. T7 RNAP in producing integral RNA (10 kb) from a linearized plasmid DNA template.Collectively, these findings establish a purification process platform for the com. production of T7 RNAP enzyme to support the future development of mRNA-based vaccines and therapeutics.