TY - JOUR
T1 - Baculovirus affinity removal in viral-based bioprocesses
AU - Moleirinho, Mafalda G.
AU - Fernandes, Rita P.
AU - Carvalho, Sofia B.
AU - Bezemer, Sandra
AU - Detmers, Frank
AU - Hermans, Pim
AU - Silva, Ricardo J. S.
AU - Alves, Paula M.
AU - Carrondo, Manuel J. T.
AU - Peixoto, Cristina
PY - 2020/6/15
Y1 - 2020/6/15
N2 - The demand for new biopharmaceuticals has grown considerably in the last decade, with their use in applications such as vaccination, gene therapy and oncolytic therapy. However, their biological complexity brings new challenges for up- and downstream processes. Insect-cell based biopharmaceuticals produced using the baculovirus expression vector system (BEVS) have an extra layer of complexity due to the co-production of baculovirus particles. Their rod-shaped form and similar envelope to virus-like particles (VLPs), results in an increasing complexity to strictly discriminate between both particles. This work presents a purification strategy based on an impurity-focused affinity chromatography approach for baculovirus removal from influenza VLPs manufacturing, evaluated as vaccine candidate. Different affinity ligands were discovered by phage display towards baculovirus particles and four ligands were selected with distinct binding profiles. The four ligands (BV-2, BV-3, BV-6, BV-8) were immobilized in chromatographic resins and dynamic binding capacities of 1.64 × 1010, 4.44 × 109, 4.91 × 1010 and 1.15 × 1011 VG/mL were obtained, respectively. The impact of the residence time in the process performance was evaluated and BV-2 and BV-3 ligands showed the best results with baculovirus removals above 70% and VLPs’ recovery yields above 60%. As a proof-of-concept, these ligands were also evaluated using two different biopharmaceuticals produced by BEVS: hepatitis C VLPs and adeno-associated virus (AAV) systems. Finally, these resins proved to be robust to harsh elution and cleaning conditions, maintaining the same baculovirus removal over 20 cycles, being a powerful chromatographic tool for baculovirus removal from biopharmaceuticals produced using BEVS.
AB - The demand for new biopharmaceuticals has grown considerably in the last decade, with their use in applications such as vaccination, gene therapy and oncolytic therapy. However, their biological complexity brings new challenges for up- and downstream processes. Insect-cell based biopharmaceuticals produced using the baculovirus expression vector system (BEVS) have an extra layer of complexity due to the co-production of baculovirus particles. Their rod-shaped form and similar envelope to virus-like particles (VLPs), results in an increasing complexity to strictly discriminate between both particles. This work presents a purification strategy based on an impurity-focused affinity chromatography approach for baculovirus removal from influenza VLPs manufacturing, evaluated as vaccine candidate. Different affinity ligands were discovered by phage display towards baculovirus particles and four ligands were selected with distinct binding profiles. The four ligands (BV-2, BV-3, BV-6, BV-8) were immobilized in chromatographic resins and dynamic binding capacities of 1.64 × 1010, 4.44 × 109, 4.91 × 1010 and 1.15 × 1011 VG/mL were obtained, respectively. The impact of the residence time in the process performance was evaluated and BV-2 and BV-3 ligands showed the best results with baculovirus removals above 70% and VLPs’ recovery yields above 60%. As a proof-of-concept, these ligands were also evaluated using two different biopharmaceuticals produced by BEVS: hepatitis C VLPs and adeno-associated virus (AAV) systems. Finally, these resins proved to be robust to harsh elution and cleaning conditions, maintaining the same baculovirus removal over 20 cycles, being a powerful chromatographic tool for baculovirus removal from biopharmaceuticals produced using BEVS.
KW - Affinity chromatography
KW - Baculovirus
KW - Downstream processing
KW - Purification
KW - Viral vectors
KW - Virus-like particles
UR - http://www.scopus.com/inward/record.url?scp=85079534046&partnerID=8YFLogxK
U2 - 10.1016/j.seppur.2020.116693
DO - 10.1016/j.seppur.2020.116693
M3 - Article
AN - SCOPUS:85079534046
VL - 241
JO - Separation and Purification Technology
JF - Separation and Purification Technology
SN - 1383-5866
M1 - 116693
ER -