Economic evaluation of M13 bacteriophage production at large‐Scale for therapeutic applications using aqueous Two‐Phase systems

Abstract

BACKGROUND: Bacteriophages are bionanoparticles with several applications in different biotechnology-based products. Among them, vaccines have the potential to treat antibiotic-resistant bacteria and parasitic infections. Traditional methods for their recovery and purification rely on precipitation with polyethylene glycol (PEG) and NaCl. However, the applicability of such an approach is limited, due to large-scale technical constrains. Recently, our research group developed a bacteriophage M13 recovery and purification strategy using Aqueous Two-Phase Systems (ATPS), simplifying the methodology and, potentially, reducing costs. This work aims to develop an economic contrast between ATPS and the traditional PEG precipitation method at different operation scales (10 to 1000 L bioreactor volume) to determine the applicability of the ATPS methodology at large scale. For this, the effect of ATPS volume ratio (V R), sample loading, and materials discount over production cost were analyzed. RESULTS: Results indicate that as the discount on material costs increases, ATPS becomes a more affordable unit operation, from a bioreactor scale of 10 L at 0% discount to 410 L at 90% discount (US$ 52.2 to $1.72 per gram, respectively). Material cost contribution is a parameter that needs attention when working with ATPS, as it is the core for the system construction. Methods for reducing their contribution are highly relevant and should be further investigated. CONCLUSION: Employment of economic analyses help to discover critical parameters for a bioprocess, such as material costs for ATPS. This economic analysis work serves as a platform for new strategies for the recovery of bacteriophage and bacteriophage-like particles using ATPS-based technologies.

Comparative Economic Analysis Between Endogenous and Recombinant Production of Hyaluronic Acid

“…An additional study was conducted to understand the impact of production titer, bioreactor scale and recovery yield. A series of Monte Carlo simulations ( Torres-Acosta et al, 2020 ; Farid, 2007 ; Torres-Acosta et al, 2018 ) were performed in which the production titer and bioreactor scale were able to change randomly in the ranges of 0.1X to 10X multipliers and 1 L to 1,000 L, respectively. For each of the 175 combinations generated, simulations were run at all the overall recovery yields calculated (0.02–66%) and the CoG/g recorded in each case.…”

“…In this study, materials are related to fermentation media and HA recovery/purification, and their costs were obtained from Sigma-Aldrich, assuming purchase of the largest presentation available (which correlated to the least expensive price per unit of mass). This approach allowed an overestimation of the material costs and, in order to stay above the real cost and prevent underestimation ( Torres-Acosta et al, 2020 ), a discount was also included in the analysis in order to reflect the reduction in prices for bulk acquisition implicit in the larger scales analyzed here. Consumables are primarily related to filtration steps and their costs were obtained from the Biosolve Process database.…”

“…This sensitivity analysis allowed determination of the impact of each variable on the CoG/g, after which the three variables with the highest contribution were varied randomly (Monte Carlo simulations) and simultaneously in order to obtain the overall possible range of CoG/g under the scenarios created ( Farid, 2007 ). To summarize this data, a multiple linear regression was constructed in the software R ( Torres-Acosta et al, 2020 ). A final comparison of the two production sources was also performed to determine the scenarios under which recombinant production could be less expensive than endogenous HA production.…”

Bacteriophage-Based Vaccines: A Potent Approach for Antigen Delivery

“…According to Munira et al the cost of producing recombinant vaccines is estimated on average of 2 USD per dose [ 84 ]. On the other hand, in, Torres-Acosta et al estimated the production cost for a phage-based vaccine was around $0.9 USD per dose (10 12 pfu) [ 85 ]. These estimations suggest a 55% reduction on the production costs of phage-based vaccines compared to recombinant-based vaccines, supporting the implementation of phages in vaccine development strategies.…”

Evaluation of the Immune Response of a Candidate Phage-Based Vaccine against Rhipicephalus microplus (Cattle Tick)

“…A recombinant phage-displaying the epitopes of Bm86 protein [21] was developed. Sbm7462 antigen is formed by the continuous fusion of three inferred immunogenic epitopes of Bm86 protein: peptide sequences 4822 (a.a. 398-411), 4823 (a.a. [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35] and 4824 (a.a. residues 132-145) (Figure 1B). A hemagglutinin (HA) tag was also inserted to facilitate detection using an anti-HA antibody.…”