{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T19:48:23Z","timestamp":1780429703574,"version":"3.54.1"},"reference-count":46,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,1,24]],"date-time":"2024-01-24T00:00:00Z","timestamp":1706054400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"AiF Project GmbH","award":["ZIM-KK5231501AJ0"],"award-info":[{"award-number":["ZIM-KK5231501AJ0"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Membranes"],"abstract":"<jats:p>New and highly selective stationary phases for affinity membrane chromatography have the potential to significantly enhance the efficiency and specificity of therapeutic protein purification by reduced mass transfer limitations. This work developed and compared different immobilization strategies for recombinant Protein A ligands to a gold-sputtered polymer membrane for antibody separation in terms of functionalization and immobilization success, protein load, and stability. Successful, functionalization was validated via X-ray photoelectron spectroscopy (XPS). Here, a recombinant Protein A ligand was coupled by N-hydroxysuccinimide (NHS)\/N-(3-dimethylaminopropyl)-N\u2032-ethylcarbodiimide (EDC) chemistry to carboxy-functionalized, gold-sputtered membranes. We achieved a binding capacity of up to 104 \u00b1 17 mg of the protein ligand per gram of the gold-sputtered membrane. The developed membranes were able to successfully capture and release the monoclonal antibody (mAb) Trastuzumab, as well as antibodies from fresh frozen human blood plasma in both static and dynamic setups. Therefore, they demonstrated successful functionalization and immobilization strategies. The antibody load was tested using bicinchoninic acid (BCA), ultraviolet-visible spectroscopy (UV-vis) measurements, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The outcome is a fully functional affinity membrane that can be implemented in a variety of different antibody purification processes, eliminating the need for creating individualized strategies for modifying the surface to suit different substrates or conditions.<\/jats:p>","DOI":"10.3390\/membranes14020031","type":"journal-article","created":{"date-parts":[[2024,1,24]],"date-time":"2024-01-24T07:42:16Z","timestamp":1706082136000},"page":"31","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Development of a New Affinity Gold Polymer Membrane with Immobilized Protein A"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5383-3538","authenticated-orcid":false,"given":"Tobias","family":"Steegm\u00fcller","sequence":"first","affiliation":[{"name":"Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstra\u00dfe 15, 85748 Garching, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8402-6640","authenticated-orcid":false,"given":"Tim","family":"Kratky","sequence":"additional","affiliation":[{"name":"Associate Professorship Physical Chemistry with Focus on Catalysis, TUM School of Natural Sciences, Technical University of Munich, Lichtenbergstra\u00dfe 4, 85748 Garching, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lena","family":"Gollwitzer","sequence":"additional","affiliation":[{"name":"Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstra\u00dfe 15, 85748 Garching, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8627-0807","authenticated-orcid":false,"given":"Sebastian Patrick","family":"Schwaminger","sequence":"additional","affiliation":[{"name":"Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstra\u00dfe 15, 85748 Garching, Germany"},{"name":"Division of Medicinal Chemistry, Otto-Loewi Research Center, Medical University of Graz, Neue Stiftingtalstra\u00dfe 6, 8010 Graz, Austria"},{"name":"BioTechMed-Graz, Mozartgasse 12, 8010 Graz, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4943-848X","authenticated-orcid":false,"given":"Sonja","family":"Berensmeier","sequence":"additional","affiliation":[{"name":"Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstra\u00dfe 15, 85748 Garching, Germany"},{"name":"Munich Institute of Integrated Materials, Energy and Process Engineering, Technical University of Munich, Lichtenbergstra\u00dfe 4a, 85748 Garching, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0021-9673(02)00057-2","article-title":"Protein separation using membrane chromatography: Opportunities and challenges","volume":"952","author":"Ghosh","year":"2002","journal-title":"J. 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