Toward QbD Process Understanding on DNA Vaccine Purification Using Design of Experiment.

Lalintip Hocharoen,Panit Kitsubun,Sarawuth Noppiboon

doi:10.3389/fbioe.2021.657201

Abstract

DNA vaccines, the third generation of vaccines, are a promising therapeutic option for many diseases as they offer the customization of their ability on protection and treatment with high stability. The production of DNA vaccines is considered rapid and less complicated compared to others such as mRNA vaccines, viral vaccines, or subunit protein vaccines. However, the main issue for DNA vaccines is how to produce the active DNA, a supercoiled isoform, to comply with the regulations. Our work therefore focuses on gaining a process understanding of the purification step which processes parameters that have impacts on the critical quality attribute (CQA), supercoiled DNA and performance attribute (PA), and step yield. Herein, pVax1/lacZ was used as a model. The process parameters of interest were sample application flow rates and salt concentration at washing step and at elution step in the hydrophobic interaction chromatography (HIC). Using a Design of Experiment (DoE) with central composite face centered (CCF) approach, 14 experiments plus four additional runs at the center points were created. The response data was used to establish regression predictive models and simulation was conducted in 10,000 runs to provide tolerance intervals of these CQA and PA. The approach of this process understanding can be applied for Quality by Design (QbD) on other DNA vaccines and on a larger production scale as well.

Highlights

DNA vaccines become prominent for use against several diseases including VCL-CB01, a candidate for cytomegalovirus (CMV)-DNA vaccine (Wloch et al, 2008), GX188E and VGX-3100 for Human Papillomavirus (HPV) (Cheng et al, 2018), a prime/boost of DNA.Mel3 with MVA.Mel3 for advanced metastatic melanoma cancer treatment (Dangoor et al, 2010), a pDERMATT for vaccination against melanoma (Quaak et al, 2008), and recently the INO-4800 for SARS-CoV2 in COVID-19 vaccine development (Smith et al, 2020)
We presented a systematic approach onto process understanding of a hydrophobic interaction chromatography (HIC)-step purification of DNA vaccine using pVax1/lacZ and Design of Experiment (DoE) as a tool
Consumables such as liquid filters were from Sartorius while Tangential Flow Filtration (TFF) cassette was from Pall. 8-mL DEAE, 8-mL C4, and 0.3 mL analytical columns were from BIA separations

Summary

Introduction

DNA vaccines become prominent for use against several diseases including VCL-CB01, a candidate for cytomegalovirus (CMV)-DNA vaccine (Wloch et al, 2008), GX188E and VGX-3100 for Human Papillomavirus (HPV) (Cheng et al, 2018), a prime/boost of DNA.Mel with MVA.Mel for advanced metastatic melanoma cancer treatment (Dangoor et al, 2010), a pDERMATT for vaccination against melanoma (Quaak et al, 2008), and recently the INO-4800 for SARS-CoV2 in COVID-19 vaccine development (Smith et al, 2020). A systematic approach, Quality by Design (QbD) developed by Juran (1992), has been introduced to pharmaceutical industries

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Results