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Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics
Thesis ( MEng)--Stellenbosch University, 2026.
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| Format: | Thesis |
| Language: | English |
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Stellenbosch : Stellenbosch University
2026
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| _version_ | 1867613888588021760 |
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| access_status_str | Open Access |
| author | Nyikadzawanda, Hudson |
| author2 | Gorgens, Johann F. |
| author_browse | Gorgens, Johann F. Nyikadzawanda, Hudson |
| author_facet | Gorgens, Johann F. Nyikadzawanda, Hudson |
| author_sort | Nyikadzawanda, Hudson |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis ( MEng)--Stellenbosch University, 2026. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/136032 |
| institution | Stellenbosch University (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:43:18.087Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2026 |
| publishDateRange | 2026 |
| publishDateSort | 2026 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/136032 Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics Nyikadzawanda, Hudson Gorgens, Johann F. Stellenbosch University. Faculty of Engineering. Dept. of Chemical Engineering. Thesis ( MEng)--Stellenbosch University, 2026. Nyikadzawanda, H. 2026. Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics. Unpublished masters thesis. Stellenbosch: Stellenbosch University [online]. Available: https://scholar.sun.ac.za/items/581d9e29-b4c3-4785-998f-46509c489e45 Africa’s near-total dependence on imported GMP-grade recombinant enzymes for mRNA vaccine production represents a critical vulnerability in continental health security, exposed most acutely during the SARS-CoV-2 pandemic. This thesis provides a framework for estimating the financial viability of manufacturing six essential mRNA vaccine-associated enzymes, T7 RNA polymerase (T7RNAP), RNase Inhibitor (RI), DNase I, Vaccinia Capping Enzyme (VCE), Inorganic Pyrophosphatase (iPPase), and 2’-O-methyltransferase (2OMT), commercially in South Africa under GMP conditions. A four-phase techno-economic methodology was employed, integrating bioprocess simulation in SuperPro Designer with capital and operating expenditure modelling, deterministic sensitivity analysis, and stochastic risk quantification through Monte Carlo simulation. Process models were constructed from mass and energy balances at production scales of 10–100 L working volume, calibrated to South African utility costs, labour market conditions, and regulatory requirements. For RI, a direct host-system comparison between Escherichia coli (E. coli) and Pichia pastoris (P. pastoris) was performed to evaluate the economic significance of microbial platform choice. The estimated costs of production and CAPEX requirements suggest that a viable and sustainable local enzyme manufacturing industry is possible, but with important limitations. Total capital investments range from USD 1.1 million for T7RNAP in E. coli to USD 3.4 million for RI in E. coli, driven by upstream cultivation scales and downstream purification complexities. Annual OPEX varies from USD 0.36 million to USD 2.34 million, yielding unit manufacturing costs (COGM) of USD 102–1 182 per mg when normalized to output. Low-titre enzymes like DNase I (5 mg/L) and VCE (10 mg/L) incur the highest burdens due to high solvent loads and resource-intensive processing, while higher-titre processes (e.g., RI at 25 mg/L) demonstrate scale economies. Enzyme contributions to facility costs highlight DNase I and RI (E. coli) as dominant (23–31%), with equipment and utilities comprising 40–70% of CAPEX, underscoring GMP operational demands. Minimum selling prices for three of the six enzymes (T7RNAP, RI, and VCE) fall within the competitive range for emerging-market GMP facilities when import logistics premiums are included. However, low-titre enzymes such as DNase I remain economically non-competitive without substantial upstream productivity improvements. Titre sensitivity analysis, varying from 0.5× to 5.0× baseline, illustrates non-linear MSP reductions: twofold improvements halve costs for low-titre targets (e.g., DNase I from USD 1 182 to 591/mg), while high-titre enzymes show diminishing returns of less than 20% reduction. MSPs after realistic titre improvements (USD 74–591/mg) position local production competitively against imported GMP enzymes (bulk prices USD 5–100/mg plus 50–300% logistics premiums), validating viability for regional supply. Feasibility and risk assessments incorporate global sensitivity, discount rates (0–50%), and Monte Carlo simulations to quantify uncertainty in inputs like capital estimates, raw materials, and titres. Production volume emerges as the primary cost driver, with +20% increases paradoxically elevating unit costs (up to +150% for DNase I) due to scale inefficiencies. Direct manufacturing costs show enzyme-specific sensitivities (–4.2% to –11.8% for –20% reductions), highlighting vulnerabilities to supply chain volatility, particularly for P. pastoris RI's methanol and oxygen demands. Discount rate analysis reveals break-even thresholds of 33–37%, with VCE demonstrating the highest resilience (37%) despite operational sensitivities, attributed to premium pricing and margins. Stochastic simulations confirm RI (P. pastoris) as most viable (NPV peaking at USD 0.5 million, payback 3–4 years, MSP ~USD 125/mg), while DNase I and T7RNAP exhibit non-viability under uncertainty, with negative NPVs and extended paybacks (>20 years), underscoring liquidity risks in emerging contexts. Host comparison for RI indicates negligible differences in minimum selling price (MSP ≈ USD 69/mg for an acceptable return on investment) at equivalent titres and relative downstream configurations. This finding, while limited to a single enzyme by biological constraints on the other five targets, emphasizes that enzyme-specific factors such as titre, localization, and downstream processing complexity outweigh microbial platform choice in determining economic viability. The findings affirm the economic potential of localized GMP enzyme manufacturing for mRNA vaccine security in Africa, with realistic 2–3× titre enhancements enabling cost-competitiveness. Recommendations prioritize upstream productivity improvements (e.g., high-cell-density fermentation, chaperone co-expression) for high-leverage targets like DNase I and VCE, alongside downstream intensification (e.g., continuous chromatography) for others. This integrated deterministic-stochastic framework bridges operational efficiency with probabilistic financial outcomes, guiding strategic investments to mitigate import dependencies and foster sustainable biomanufacturing ecosystems. Masters 2026-04-21T07:21:47Z 2026-04-21T07:21:47Z 2026-03 Thesis https://scholar.sun.ac.za/handle/10019.1/136032 en Stellenbosch University application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Nyikadzawanda, Hudson Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title | Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title_full | Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title_fullStr | Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title_full_unstemmed | Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title_short | Manufacturing GMP-grade mRNA vaccine-associated Enzymes: Bioprocess Development and Economics |
| title_sort | manufacturing gmp grade mrna vaccine associated enzymes bioprocess development and economics |
| url | https://scholar.sun.ac.za/handle/10019.1/136032 |
| work_keys_str_mv | AT nyikadzawandahudson manufacturinggmpgrademrnavaccineassociatedenzymesbioprocessdevelopmentandeconomics |