{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T13:54:12Z","timestamp":1790085252479,"version":"4.0.1"},"reference-count":37,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T00:00:00Z","timestamp":1779926400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T00:00:00Z","timestamp":1779926400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100000781","name":"European Research Council","doi-asserted-by":"publisher","award":["101158248\u2010UNMASK\u2010ERC\u20102023\u2010POC"],"award-info":[{"award-number":["101158248\u2010UNMASK\u2010ERC\u20102023\u2010POC"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Recombinant Staphylococcal Protein A is the most widely used affinity ligand for industrial antibody purification, as well as for research and discovery of new antibodies. It has become a platform technology throughout the antibody manufacturing industry for the direct capture of products from a clarified cell culture supernatant. However, protein A adsorbents contribute significantly to manufacturing costs in the early stage of process development. In this work, we aimed to increase the production yield of an engineered protein A ligand comprising six repeats of the alkali\u2010resistant protein A\u2010derived Z domain, with a terminal cysteine residue for adsorbent immobilization (Z6AlkC). We coupled the extracellular production of Z6AlkC via secretion in high\u2010cell\u2010density, carbon\u2010limited bioreactor cultures, with a simple and fast ultradiafiltration step for purification. Our bioprocess achieved 4\u00a0g\/L of secreted Z6AlkC with an 89% purity, which is an improvement from previous reports that obtained intracellular expression and lower yield. We further purified the Z6AlkC protein by IgG\u2010based affinity chromatography (Z6AlkC\u2010P) and ultradiafiltration (Z6AlkC\u2010F) and used it as an affinity ligand for antibody purification, as a proof of functionality. Both ligands yielded similar purification antibody behavior, showing that high\u2010performance protein A\u2010based ligands can be produced in scalable, high\u2010productivity, and cost\u2010effective methods.<\/jats:p>","DOI":"10.1002\/biot.70247","type":"journal-article","created":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T08:01:16Z","timestamp":1779955276000},"source":"Crossref","is-referenced-by-count":1,"title":["High\u2010Yield Production and Cost\u2010Effective Purification of an Alkali\u2010Resistant Hexameric Protein A Variant for Antibody Affinity Chromatography"],"prefix":"10.1002","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5185-9294","authenticated-orcid":false,"given":"Carolina Mota","family":"Natal","sequence":"first","affiliation":[{"name":"Associate Laboratory i4HB \u2013 Institute for Health and Bioeconomy Department of Chemistry, NOVA School of Science and Technology Universidade NOVA de Lisboa Caparica Portugal"},{"name":"UCIBIO \u2013 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