{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T10:09:35Z","timestamp":1784196575562,"version":"3.55.0"},"reference-count":116,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T00:00:00Z","timestamp":1784160000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T00:00:00Z","timestamp":1784160000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000038","name":"U.S. Food and Drug Administration","doi-asserted-by":"publisher","award":["R01FD007481"],"award-info":[{"award-number":["R01FD007481"]}],"id":[{"id":"10.13039\/100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014726","name":"National Institute for Innovation in Manufacturing Biopharmaceuticals","doi-asserted-by":"publisher","award":["PC3.1\u2010119"],"award-info":[{"award-number":["PC3.1\u2010119"]}],"id":[{"id":"10.13039\/100014726","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014726","name":"National Institute for Innovation in Manufacturing Biopharmaceuticals","doi-asserted-by":"publisher","award":["PC5.2\u2010133"],"award-info":[{"award-number":["PC5.2\u2010133"]}],"id":[{"id":"10.13039\/100014726","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014726","name":"National Institute for Innovation in Manufacturing Biopharmaceuticals","doi-asserted-by":"publisher","award":["PC8.1\u2010135"],"award-info":[{"award-number":["PC8.1\u2010135"]}],"id":[{"id":"10.13039\/100014726","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Modern biopharmaceutical manufacturing requires purification platforms capable of processing structurally and functionally diverse products while addressing the challenge of removing persistent and high\u2010risk host cell proteins (HCPs). Current purification processes utilize adsorbents that operate in bind\u2010and\u2010elute mode, which limits productivity, and often struggle to remove critical impurities. Addressing these limitations, our team developed a flow\u2010through pseudo\u2010affinity chromatography platform that utilizes size\u2010exclusion resins (Monomix Core 500) functionalized with mixed\u2010mode peptide ligands (AviGuard) to remove process\u2010related impurities while maximizing productivity and product integrity. This study evaluates this technology across three therapeutic modalities: two bi\u2010specific monoclonal antibodies (bsAbs) and an Fc\u2010fusion protein from Chinese hamster ovary (CHO) cell culture fluids, and adeno\u2010associated virus serotypes 2 and 9 (AAV2 and AAV9) from HEK293 cell lysates. For the CHO\u2010derived proteins, the platform comprised a pre\u2010capture step with LigaGuard\u2010Monomix Core 500 resin (LGMC500), a Protein A\u2010based capture step, and a polishing step using a single\u2010use size\u2010exclusion mixed\u2010mode (SEMM) resin. This configuration achieved global product yields exceeding 70%, final product pool concentration &gt;\u200915\u2009mg\/mL, monomeric purity ~99%, and excellent cumulative HCP clearance (LRV\u2009&gt;\u20094.5) to residual levels of 60\u2013165\u2009ng\/mL (4\u201311\u2009ppm). For AAV purification, the process utilized a mixed\u2010bed adsorbent comprising dextran\u2010coated charcoal and LGMC500 for removing HCPs, cell metabolites, and media components, followed by a capture step using either CaptureSelect AAVX resin or Natrix CH membranes. The optimization of the charcoal\u2010to\u2010resin volume ratio (1:3) and load volume (~\u200910\n                    <jats:sup>14<\/jats:sup>\n                    vp per mL of resin) yielded ~50% AAV recovery with a residual HCP level of 350\u2009ng\/mL, representing &lt;\u2009100\u2009ng per dose, in compliance with regulatory guidelines. Process productivity ranged from 19 to 23\u2009g\/L\u00b7hr for CHO\u2010derived proteins and ~7\u2009\u00d7\u200910\n                    <jats:sup>14<\/jats:sup>\n                    vp\/L\u00b7hr for AAV9, positioning this technology in the optimal operating space for modern bioprocessing.\n                  <\/jats:p>","DOI":"10.1002\/bit.70310","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T09:39:01Z","timestamp":1784194741000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Purification of Advanced Therapeutics by Flow\u2010Through Pseudo\u2010Affinity Chromatography: Bi\u2010Specific mABs, Fc Fusion Proteins, and Adeno\u2010Associated Viruses"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6033-0963","authenticated-orcid":false,"given":"Wenning","family":"Chu","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eduardo","family":"Barbieri","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4504-9567","authenticated-orcid":false,"given":"Arianna","family":"Minzoni","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"},{"name":"North Carolina Viral Vector Initiative in Research and Learning (NC\u2010VVIRAL) NC State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shriarjun","family":"Shastry","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) NC State University Raleigh North Carolina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob P.","family":"LeBarre","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingbo","family":"Shu","sequence":"additional","affiliation":[{"name":"Molecular Education, Technology, and Research Innovation Center (METRIC) NC State university Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taufika","family":"Williams","sequence":"additional","affiliation":[{"name":"Molecular Education, Technology, and Research Innovation Center (METRIC) NC State university Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7548-583X","authenticated-orcid":false,"given":"Xuanwen","family":"Li","sequence":"additional","affiliation":[{"name":"Analytical Research &amp; Development Merck &amp; Co. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3676-7209","authenticated-orcid":false,"given":"Divya","family":"Chandra","sequence":"additional","affiliation":[{"name":"Process Research and Development Merck &amp; Co. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuankuo","family":"Xu","sequence":"additional","affiliation":[{"name":"Bristol Myers Squibb Devens Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerome","family":"Bill","sequence":"additional","affiliation":[{"name":"Genentech Inc., 1 DNA Way South San Francisco California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Roush","sequence":"additional","affiliation":[{"name":"Roush Biopharma Panacea Colts Neck New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Taris","sequence":"additional","affiliation":[{"name":"ChromaGenix Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Gilbert","sequence":"additional","affiliation":[{"name":"ChromaGenix Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andre","family":"Kiesewetter","sequence":"additional","affiliation":[{"name":"Merck Life Science KGaA Darmstadt Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oliver","family":"Rammo","sequence":"additional","affiliation":[{"name":"Merck Life Science KGaA Darmstadt Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Schulte","sequence":"additional","affiliation":[{"name":"Merck Life Science KGaA Darmstadt Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher","family":"Major","sequence":"additional","affiliation":[{"name":"ChromaGenix Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trina","family":"Mouchahoir","sequence":"additional","affiliation":[{"name":"Biomolecular Measurement Division National Institute of Standards and Technology Gaithersburg Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4993-0826","authenticated-orcid":false,"given":"Steven","family":"Cramer","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering Rensselaer Polytechnic Institute Troy New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiana","family":"Boi","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) NC State University Raleigh North Carolina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruben G.","family":"Carbonell","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) NC State University Raleigh North Carolina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7831-219X","authenticated-orcid":false,"given":"Abraham","family":"Lenhoff","sequence":"additional","affiliation":[{"name":"North Carolina Viral Vector Initiative in Research and Learning (NC\u2010VVIRAL) NC State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5633-434X","authenticated-orcid":false,"given":"Stefano","family":"Menegatti","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering NC State University Raleigh North Carolina USA"},{"name":"North Carolina Viral Vector Initiative in Research and Learning (NC\u2010VVIRAL) NC State University Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) NC State University Raleigh North Carolina"},{"name":"ChromaGenix Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,16]]},"reference":[{"issue":"6","key":"e_1_2_8_2_1","doi-asserted-by":"crossref","DOI":"10.1002\/btm2.70017","article-title":"Developing a Process of Lentivirus Purification From Recombinant Fluids Using Peptide Affinity Ligands","volume":"10","author":"Barbieri E.","year":"2025","journal-title":"Bioengineering & Translational Medicine"},{"issue":"5","key":"e_1_2_8_3_1","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1002\/btpr.2150","article-title":"Trace Levels of the CHO Host Cell Protease Cathepsin D Caused Particle Formation in a Monoclonal Antibody Product","volume":"31","author":"Bee J. 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