{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T11:05:14Z","timestamp":1784804714521,"version":"3.55.0"},"reference-count":71,"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:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"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"}],"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                    Countercurrent dialysis using hollow fiber modules offers several advantages over both inline diafiltration (ILDF) and hemodialyzer based countercurrent dialysis in terms of buffer requirements and compatibility with GMP manufacturing of biologics. In this work, we evaluate GMP\u2010grade hollow fiber modules of varying lengths (20, 41.5, and 65\u2009cm) and benchmark their performance against an ILDF unit as a reference process for continuous exchange of buffer in manufacturing of monoclonal antibodies. Hollow fiber modules outperformed the ILDF configuration in terms of buffer consumption and pressure build\u2010up, particularly for higher\u2010titer processes. The GMP hollow fiber modules, which are characterized using protein\u2010based molecular weight cut\u2010off (MWCO) specifications, achieved buffer exchange comparable to the reference processes when using endogenous excipients (histidine, methionine) as tracers, while providing a bioprocess\u2010oriented format and documentation suitable for GMP implementation. A D\u2010optimal experimental design was used to define the design space for the hollow fiber countercurrent dialysis step by varying the ratio of dialysate buffer to feed flow (\u201calpha factor\u201d, 2\u20135), normalized feed flow rate (0.05\u20131.5\u2009L\u00b7m\n                    <jats:sup>\u22122<\/jats:sup>\n                    \u00b7h\n                    <jats:sup>\u22121<\/jats:sup>\n                    ), protein concentration (30\u201370\u2009mg\/mL), and feed conductivity (2\u201315\u2009mS\/cm). The results showed that higher feed concentrations narrowed the acceptable range of normalized feed flow rates and required higher alpha factors to achieve the target buffer exchange. A 5\u2010day continuous run operated at the selected process set\u2010point achieved the desired buffer exchange targets with &gt;\u200995% product yield, no significant impact on aggregation, and transmembrane pressures below 12\u2009psi (\u224880\u2009kPa). Scale\u2010up analysis indicated that the process is suitable for GMP manufacturing at scales up to 3000\u2009L perfusion bioreactors with perfusate titers ranging from 0.6 to 5\u2009mg\/mL, while operating within a robust design space.\n                  <\/jats:p>","DOI":"10.1002\/bit.70313","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T09:47:32Z","timestamp":1784195252000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Optimization of a Hollow Fiber\u2010Based Countercurrent Dialysis Process for Continuous Manufacturing of Monoclonal Antibodies"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3291-7345","authenticated-orcid":false,"given":"Vishwanath","family":"Hebbi","sequence":"first","affiliation":[{"name":"Process Research and Development MRL, Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3277-4513","authenticated-orcid":false,"given":"Mark","family":"Brower","sequence":"additional","affiliation":[{"name":"Process Research and Development MRL, Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nuno D. S.","family":"Pinto","sequence":"additional","affiliation":[{"name":"Process Research and Development MRL, Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,16]]},"reference":[{"issue":"015","key":"e_1_2_10_2_1","first-page":"1","article-title":"Inline Diafiltration With Multi\u2010Channel Pump","volume":"15","author":"Ayturk E.","year":"2019","journal-title":"US PTO US"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1201\/9781482278743"},{"key":"e_1_2_10_4_1","volume-title":"A\u2010Mab: A Case Study in Bioprocess Development","author":"CMC\u2010Biotech Working Group","year":"2009"},{"key":"e_1_2_10_5_1","first-page":"1","article-title":"Methods for Unified Concentration and Buffer Exchange","author":"Coffman J.","year":"2022","journal-title":"US Patents"},{"key":"e_1_2_10_6_1","first-page":"44","article-title":"Hollow Fiber Cartridges for Membrane Separations: Operating Handbook","volume":"1","author":"Cytiva","year":"2022","journal-title":"Cytiva"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2006.03.011"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.memsci.2014.08.026"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.24377"},{"key":"e_1_2_10_10_1","first-page":"40","article-title":"Factors influencing ultrafiltration scale\u2010upInfluencing Ultrafiltration Scale\u2010Up","volume":"3","author":"Dosmar M.","year":"2005","journal-title":"BioProcess International"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4613-2313-6_12"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/9781119126942.ch21"},{"key":"e_1_2_10_13_1","unstructured":"Fernandez Cerezo L. 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