Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies
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| Title: | Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies |
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| Authors: | Ines Zimmermann, Yasmin Kaveh-Baghbaderani, Friederike Eilts, Nadja Kohn, Paula Fraga-García, Sonja Berensmeier |
| Source: | ACS Appl Bio Mater |
| Publisher Information: | American Chemical Society (ACS), 2024. |
| Publication Year: | 2024 |
| Subject Terms: | magnetic nanoparticles, Staphylococcal Protein A/chemistry [MeSH], Immunoglobulin G/chemistry [MeSH], downstream processing, pH buffering of iron oxides, Magnetic Iron Oxide Nanoparticles/chemistry [MeSH], kinetics, Biocompatible Materials/chemistry [MeSH], Article, Ligands [MeSH], Surface Properties [MeSH], Immunoglobulin G/isolation, Antibodies, Monoclonal/chemistry [MeSH], Staphylococcal Protein A/metabolism [MeSH], Materials Testing [MeSH], Particle Size [MeSH], protein recovery, Antibodies, Monoclonal/isolation, site-directed Protein A immobilization, Ferric Compounds/chemistry [MeSH], Surface Properties, Immunoglobulin G, Materials Testing, Antibodies, Monoclonal, Magnetic Iron Oxide Nanoparticles, Biocompatible Materials, pH buffering ofiron oxides, Particle Size, Ligands, Staphylococcal Protein A, Ferric Compounds, ddc |
| Description: | Magnetic separation is a promising alternative to chromatography for enhancing the downstream processing (DSP) of monoclonal antibodies (mAbs). However, there is a lack of efficient magnetic particles for successful application. Aiming to fill this gap, we demonstrate the suitability of bare iron oxide nanoparticles (BION) with physical site-directed immobilization of an engineered Protein A affinity ligand (rSpA) as an innovative magnetic material. The rSpA ligand contains a short peptide tag that enables the direct and stable immobilization onto the uncoated BION surface without commonly required laborious particle activation. The resulting BION@rSpA have beneficial characteristics outperforming conventional Protein A-functionalized magnetic particles: a simple, fast, low-cost synthesis, a particle size in the nanometer range with a large effective specific surface area enabling large immunoglobulin G (IgG) binding capacity, and a high magnetophoretic velocity advantageous for fast processing. We further show rapid interactions of IgG with the easily accessible rSpA ligands. The binding of IgG to BION@rSpA is thereby highly selective and not impeded by impurity molecules in perfusion cell culture supernatant. Regarding the subsequent acidic IgG elution from BION@rSpA@IgG, we observed a hampering pH increase caused by the protonation of large iron oxide surfaces after concentrating the particles in 100 mM sodium acetate buffer. However, the pH can be stabilized by adding 50 mM glycine to the elution buffer, resulting in recoveries above 85% even at high particle concentrations. Our work shows that BION@rSpA enable efficient magnetic mAb separation and could help to overcome emerging bottlenecks in DSP. |
| Document Type: | Article Other literature type |
| File Description: | application/pdf |
| Language: | English |
| ISSN: | 2576-6422 |
| DOI: | 10.1021/acsabm.4c00280 |
| Access URL: | https://pubmed.ncbi.nlm.nih.gov/38740514 https://repository.publisso.de/resource/frl:6487163 https://mediatum.ub.tum.de/doc/1747668/document.pdf |
| Rights: | CC BY URL: http://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (http://creativecommons.org/licenses/by/4.0/). |
| Accession Number: | edsair.doi.dedup.....d23fb50a8fe51a1cf22a77e5ba0ca103 |
| Database: | OpenAIRE |
Nájsť tento článok vo Web of Science | Abstract: | Magnetic separation is a promising alternative to chromatography for enhancing the downstream processing (DSP) of monoclonal antibodies (mAbs). However, there is a lack of efficient magnetic particles for successful application. Aiming to fill this gap, we demonstrate the suitability of bare iron oxide nanoparticles (BION) with physical site-directed immobilization of an engineered Protein A affinity ligand (rSpA) as an innovative magnetic material. The rSpA ligand contains a short peptide tag that enables the direct and stable immobilization onto the uncoated BION surface without commonly required laborious particle activation. The resulting BION@rSpA have beneficial characteristics outperforming conventional Protein A-functionalized magnetic particles: a simple, fast, low-cost synthesis, a particle size in the nanometer range with a large effective specific surface area enabling large immunoglobulin G (IgG) binding capacity, and a high magnetophoretic velocity advantageous for fast processing. We further show rapid interactions of IgG with the easily accessible rSpA ligands. The binding of IgG to BION@rSpA is thereby highly selective and not impeded by impurity molecules in perfusion cell culture supernatant. Regarding the subsequent acidic IgG elution from BION@rSpA@IgG, we observed a hampering pH increase caused by the protonation of large iron oxide surfaces after concentrating the particles in 100 mM sodium acetate buffer. However, the pH can be stabilized by adding 50 mM glycine to the elution buffer, resulting in recoveries above 85% even at high particle concentrations. Our work shows that BION@rSpA enable efficient magnetic mAb separation and could help to overcome emerging bottlenecks in DSP. |
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| ISSN: | 25766422 |
| DOI: | 10.1021/acsabm.4c00280 |