TY - JOUR
T1 - Membrane-Based Approach for the Downstream Processing of Influenza Virus-Like Particles
AU - Carvalho, Sofia B.
AU - Silva, Ricardo J.S.
AU - Moleirinho, Mafalda G.
AU - Cunha, Bárbara
AU - Moreira, Ana S.
AU - Xenopoulos, Alex
AU - Alves, Paula M.
AU - Carrondo, Manuel J.T.
AU - Peixoto, Cristina
PY - 2019/8/1
Y1 - 2019/8/1
N2 - Currently, marketed influenza vaccines are only efficient against homologous viruses, thus requiring a seasonal update based on circulating subtypes. This constant reformulation adds several challenges to manufacturing, particularly in purification due to the variation of the physicochemical properties of the vaccine product. A universal platform approach capable of handling such variation is therefore of utmost importance. In this work, a filtration-based approach is explored to purify influenza virus-like particles. Switching from adsorptive separation to size-based purification allows overcoming the differences in retention observed for different influenza strains. The proposed process employs a cascade of ultrafiltration and diafiltration steps, followed by a sterile filtration step. Different process parameters are assessed in terms of product recovery and impurities’ removal. Membrane chemistry, pore size, operation modes, critical flux, transmembrane pressure, and permeate control strategies are evaluated. After membrane selection and parameter optimization, concentration factors and diafiltration volumes are also defined. By optimizing the filtration mode of operation, it is possible to achieve product recoveries of approximately 80%. Overall, the process time is decreased by 30%, its scalability is improved, and the costs are reduced due to the removal of chromatography and associated buffer consumptions, cleaning, and its validation steps.
AB - Currently, marketed influenza vaccines are only efficient against homologous viruses, thus requiring a seasonal update based on circulating subtypes. This constant reformulation adds several challenges to manufacturing, particularly in purification due to the variation of the physicochemical properties of the vaccine product. A universal platform approach capable of handling such variation is therefore of utmost importance. In this work, a filtration-based approach is explored to purify influenza virus-like particles. Switching from adsorptive separation to size-based purification allows overcoming the differences in retention observed for different influenza strains. The proposed process employs a cascade of ultrafiltration and diafiltration steps, followed by a sterile filtration step. Different process parameters are assessed in terms of product recovery and impurities’ removal. Membrane chemistry, pore size, operation modes, critical flux, transmembrane pressure, and permeate control strategies are evaluated. After membrane selection and parameter optimization, concentration factors and diafiltration volumes are also defined. By optimizing the filtration mode of operation, it is possible to achieve product recoveries of approximately 80%. Overall, the process time is decreased by 30%, its scalability is improved, and the costs are reduced due to the removal of chromatography and associated buffer consumptions, cleaning, and its validation steps.
KW - filtration
KW - influenza
KW - tangential flow filtration
KW - vaccines
KW - virus-like particles
UR - http://www.scopus.com/inward/record.url?scp=85069877680&partnerID=8YFLogxK
U2 - 10.1002/biot.201800570
DO - 10.1002/biot.201800570
M3 - Article
C2 - 31106962
AN - SCOPUS:85069877680
SN - 1860-6768
VL - 14
JO - Biotechnology Journal
JF - Biotechnology Journal
IS - 8
M1 - 1800570
ER -