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Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads

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Title: Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads
Authors: Pereira Aguilar, Patricia, Reiter, Katrin, Wetter, Viktoria, Steppert, Petra, Maresch, Daniel, Ling, Wai Li, Satzer, Peter, Jungbauer, Alois
Contributors: Universität für Bodenkultur Wien = University of Natural Resources and Life Sciences Vienne, Autriche (BOKU), Austrian Centre of Industrial Biotechnology GmbH (ACIB), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), ANR-10-LABX-0049,GRAL,Grenoble Alliance for Integrated Structural Cell Biology(2010)
Source: ISSN: 0021-9673.
Publisher Information: CCSD
Elsevier
Publication Year: 2020
Collection: Université Grenoble Alpes: HAL
Subject Terms: Affinity chromatography, Convective media, Downstream processing, Enveloped VLP, Ion exchange chromatography, Polymer-grafted media, MESH: Adsorption, MESH: Animals, MESH: CHO Cells, MESH: Chromatin, MESH: Chromatography, Affinity, MESH: Convection, MESH: Cricetinae, MESH: Cricetulus, MESH: HIV-1, MESH: Histones, MESH: Humans, MESH: Microspheres, MESH: Nanoparticles, MESH: Polymers, MESH: Porosity, MESH: Virion, MESH: Anions, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
Description: International audience ; Downstream processing (DSP) of large bionanoparticles is still a challenge. The present study aims to systematically compare some of the most commonly used DSP strategies for capture and purification of enveloped viruses and virus-like particles (eVLPs) by using the same staring material and analytical tools. As a model, Human Immunodeficiency Virus-1 (HIV-1) gag VLPs produced in CHO cells were used. Four different DSP strategies were tested. An anion-exchange monolith and a membrane adsorber, for direct capture and purification of eVLPs, and a polymer-grafted anion-exchange resin and a heparin-affinity resin for eVLP purification after a first flow-through step to remove small impurities. All tested strategies were suitable for capture and purification of eVLPs. The performance of the different strategies was evaluated regarding its binding capacity, ability to separate different particle populations and product purity. The highest binding capacity regarding total particles was obtained using the anion exchange membrane adsorber (5.3 × 1012 part/mL membrane), however this method did not allow the separation of different particle populations. Despite having a lower binding capacity (1.5 × 1011 part/mL column) and requiring a pre-processing step with flow-through chromatography, Heparin-affinity chromatography showed the best performance regarding separation of different particle populations, allowing not only the separation of HIV-1 gag VLPs from host cell derived bionanoparticles but also from chromatin. This work additionally shows the importance of thorough sample characterization combining several biochemical and biophysical methods in eVLP DSP.
Document Type: article in journal/newspaper
Language: English
Relation: info:eu-repo/semantics/altIdentifier/pmid/32823092; PUBMED: 32823092
DOI: 10.1016/j.chroma.2020.461378
Availability: https://hal.science/hal-02974741
https://hal.science/hal-02974741v1/document
https://hal.science/hal-02974741v1/file/Pereira%20et%20al.%20J%20Chromatography.pdf
https://doi.org/10.1016/j.chroma.2020.461378
Rights: info:eu-repo/semantics/OpenAccess
Accession Number: edsbas.C4F47ADF
Database: BASE
Description
Abstract:International audience ; Downstream processing (DSP) of large bionanoparticles is still a challenge. The present study aims to systematically compare some of the most commonly used DSP strategies for capture and purification of enveloped viruses and virus-like particles (eVLPs) by using the same staring material and analytical tools. As a model, Human Immunodeficiency Virus-1 (HIV-1) gag VLPs produced in CHO cells were used. Four different DSP strategies were tested. An anion-exchange monolith and a membrane adsorber, for direct capture and purification of eVLPs, and a polymer-grafted anion-exchange resin and a heparin-affinity resin for eVLP purification after a first flow-through step to remove small impurities. All tested strategies were suitable for capture and purification of eVLPs. The performance of the different strategies was evaluated regarding its binding capacity, ability to separate different particle populations and product purity. The highest binding capacity regarding total particles was obtained using the anion exchange membrane adsorber (5.3 × 1012 part/mL membrane), however this method did not allow the separation of different particle populations. Despite having a lower binding capacity (1.5 × 1011 part/mL column) and requiring a pre-processing step with flow-through chromatography, Heparin-affinity chromatography showed the best performance regarding separation of different particle populations, allowing not only the separation of HIV-1 gag VLPs from host cell derived bionanoparticles but also from chromatin. This work additionally shows the importance of thorough sample characterization combining several biochemical and biophysical methods in eVLP DSP.
DOI:10.1016/j.chroma.2020.461378