Periodic Reporting for period 3 - DiViNe (Sustainable downstream processing of vaccines through incorporation of nanobiotechnologies : novel affinity ligands and biomimetic membranes) Periodo di rendicontazione: 2018-03-01 al 2020-02-29 Sintesi del contesto e degli obiettivi generali del progetto The process of purification and recovery of biopharmaceuticals, such as vaccines, is generally referred to as “downstream processing”. This process aims at removing the maximum of contaminants while recovering as much of the therapeutic molecule as possible. Currently, this process relies in multiple «negative» selection/purification steps which negatively impact recovery yields while increasing costs and waste products. In contrast, the development of «positive» separation methods like affinity targeted in Divine, could significantly reduce the number of purification steps resulting in faster downstream processes and higher purification yields, leading to a reduction in costs and waste could also be achieved. Water recycling is a major concern in the vaccine industry, as the aforementioned purification steps result in large volumes of “unclean” water that are directed to Wastewater Treatment Plants. This use of water is of concern for developing countries, as the strategy of pharma industry to build local production plants in an attempt to reduce vaccination cost is significantly impacted by water availability. The DiViNe project is tackling these vaccine availability and cost and environmental issues with technological solutions. The consortium combines two major Nano/biotechnology innovations to develop an integrated vaccine purification platform, amenable to different types of vaccines, increasing titers, lowering costs and reducing wastewater production.The consortium is composed of Affilogic, which combines efforts with Merck to generate Nanofitin-based chromatographic columns (novel affinity capture ligands), iBET, developing affinity chromatography for enveloped vaccines and bio-processes to enable a sustainable large scale production of Nanofitins, GSK Vaccines providing the consortium with a broad range of vaccine candidates as targets for purification and who will test the performance of the novel affinity chromatography columns, Aquaporin introducing use water and energy-saving nano-biomimetics based membranes for water decontamination, and Genibet producing under cGMP the intermediate master cell banks. The custom affinity capture processes being developed as a sustainable platform is therefore economically relevant, in a very large market. Beyond the technical partnership, the project is a first run for the partners to structure the platform as a commercial offer for downstream processing of biologics in particular, biopharmaceuticals. Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti The DiViNe project targets three different families of vaccines: Family 1 (TF1): Carrier proteins used for glycoconjugate vaccines, extracted from live bacteria and for which only traditional purification technologies are applied, with low yields, Family 2 (TF2): Protein antigens that are particularly hard to separate from by products such as truncated forms, thereby requesting highly specific capture and Family 3 (TF3): Fragile enveloped viruses, not amenable to usual purification systems. In the last 5 years, Nanofitins against all targets were generated and immobilized into affinity columns or membranes. Proof of concept for the advantage of using Nanofitin-based chromatographic columns at pilot scale was generated for TF1 and TF2. In both cases, the use of Nanofitins-functionalized resins resulted in similar or higher yields of purified product, lower costs of goods and lower volume of generated wastewater. The promising results observed with TF1 and TF2 prompted the development of new Nanofitins-functionalized resins for a new vaccine candidate from TF2 family (called TF2*). This new vaccine candidate was fast-tracked as a result from GSK interest to move it into clinical testing. In the specific case of TF2*, the new chromatographic method was scaled-up beyond pilot to industrial scale. For the purification of TF3, two different and complimentary approaches were used: (i) positive selection and (ii) negative selection. In both cases, Nanofitins were generated and are being used in membrane-based chromatography systems. Genibet is completing the production under cGMP of Master Cell Banks for the most relevant Nanofitins.The complementary arm of the DiViNe project performed by Aquaporin in WP2 adapts a Forward Osmosis (FO) process for reducing volume of waste water from vaccine production downstream fermentation processes. Broths were provided by GSK for TF1 and TF2 and by iBET for TF3. Both flat sheet membranes and hollow fibre modules have been assessed. Promising results were obtained during concentration experiments of TF1 and TF2 on pilot scale using newly developed FO hollow fiber membranes, while TF3 results are being finalized. Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora) The objective of the DiViNe project is to streamline and reduce the number of purification steps of vaccines using chromatography methods and improve water usage in the process. Proof of concept paths used three different types of vaccine candidates, (i) a glycoconjugate (TF1), (ii) a Protein (TF2), and (iii) enveloped virus-like particles (TF3). The scope of this project was further enriched with the incorporation of a second member of the target family 2 (TF2*), a GSK-vaccine candidate now entering toxicology and clinical studies. The new activities with TF2* leveraged the Technology readiness levels of the novel affinity ligands and biomimetic membranes from “Development” to “Demonstration”. Moreover, this platform-approach, from target to affinity material, illustrates the custom platform that is now available for commercial service in the biopharmaceutical field at large. Such process is simple and versatile enabling the development of functionalized resins and membranes for a variety of biomolecules and therapeutic applications. Finally the forward osmosis developed proved their value in improving water usage.The progress of DiViNe will be confronted to the needs of emerging countries in terms of enhancing access to vaccines. To this end, Vito di Cioccio, from GSK Vaccines Institute for Global Health, Odile Leroy, from European Vaccine Initiative and Katey Owen, from The Bill and Melinda Gates Foundation constituted the External Experts Advisory Group, which met to assess, criticize and support the concept. concept-2.png concept-1.jpg
