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Therefore, improved manufacturing processes are needed to cope with the high number of functional particles required for clinical trials and, eventually, commercialization. Affinity chromatography (AC) can be used to simplify purification processes and generate clinical-grade products with high titer and purity. However, one of the major challenges in the purification of Lentiviral vectors (LVs) using AC is to combine a highly specific ligand with a gentle elution condition assuring the preservation of vector biological activity. In this work, we report for the first time the implementation of an AC resin to specifically purify VSV-G pseudotyped LVs. After ligand screening, different critical process parameters were assessed and optimized. A dynamic capacity of 1 \u00d7 1011 total particles per mL of resin was determined and an average recovery yield of 45% was found for the small-scale purification process. The established AC robustness was confirmed by the performance of an intermediate scale providing an infectious particles yield of 54%, which demonstrates the scalability and reproducibility of the AC matrix. Overall, this work contributes to increasing downstream process efficiency by delivering a purification technology that enables high purity, scalability, and process intensification in a single step, contributing to time-to-market reduction.<\/jats:p>","DOI":"10.3390\/ijms24043354","type":"journal-article","created":{"date-parts":[[2023,2,8]],"date-time":"2023-02-08T03:01:45Z","timestamp":1675825305000},"page":"3354","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Implementation of Novel Affinity Ligand for Lentiviral Vector Purification"],"prefix":"10.3390","volume":"24","author":[{"given":"Ana Sofia","family":"Moreira","sequence":"first","affiliation":[{"name":"IBET Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2780-901 Oeiras, Portugal"},{"name":"ITQB Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa, Avenida da Rep\u00fablica, 2780-157 Oeiras, Portugal"}]},{"given":"Sandra","family":"Bezemer","sequence":"additional","affiliation":[{"name":"Thermo Fisher Scientific, 2333 CH Leiden, The Netherlands"}]},{"given":"Tiago Q.","family":"Faria","sequence":"additional","affiliation":[{"name":"IBET Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2780-901 Oeiras, Portugal"}]},{"given":"Frank","family":"Detmers","sequence":"additional","affiliation":[{"name":"Thermo Fisher Scientific, 2333 CH Leiden, The Netherlands"}]},{"given":"Pim","family":"Hermans","sequence":"additional","affiliation":[{"name":"Thermo Fisher Scientific, 2333 CH Leiden, The Netherlands"}]},{"given":"Laurens","family":"Sierkstra","sequence":"additional","affiliation":[{"name":"Thermo Fisher Scientific, 2333 CH Leiden, The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9728-6890","authenticated-orcid":false,"given":"Ana Sofia","family":"Coroadinha","sequence":"additional","affiliation":[{"name":"IBET Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2780-901 Oeiras, Portugal"}]},{"given":"Cristina","family":"Peixoto","sequence":"additional","affiliation":[{"name":"IBET Instituto de Biologia Experimental e Tecnol\u00f3gica, Apartado 12, 2780-901 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1517\/14712598.2013.779249","article-title":"New developments in lentiviral vector design, production and purification","volume":"13","author":"Segura","year":"2013","journal-title":"Expert Opin. 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