Zitieren Sie bitte immer diesen URN: urn:nbn:de:kobv:b43-555142
High-purity corundum as support for affinity extractions from complex samples
- Nonporous corundum powder, known as an abrasive material in the industry, was functionalized covalently with protein binders to isolate and enrich specific proteins from complex matrices. The materials based on corundum were characterized by TEM, ESEM, BET, DLS, and zeta potential measurements. The strong Al-O-P bonds between the corundum surface and amino phosphonic acids are used to introduce functional groups for further conjugations. The common cross-linker glutaraldehyde was compared with a hyperbranched polyglycerol (PG) of around 10 kDa. The latter is oxidized with periodate to generate aldehyde groups that can covalently react with the amines of the surface and the amino groups from the protein via a reductive amination process. The amount of bound protein was quantified via aromatic amino acid analysis (AAAA). This work shows that oxidized polyglycerol can be used as an alternative to glutaraldehyde. With polyglycerol, more of the model protein bovine serum albumin (BSA) couldNonporous corundum powder, known as an abrasive material in the industry, was functionalized covalently with protein binders to isolate and enrich specific proteins from complex matrices. The materials based on corundum were characterized by TEM, ESEM, BET, DLS, and zeta potential measurements. The strong Al-O-P bonds between the corundum surface and amino phosphonic acids are used to introduce functional groups for further conjugations. The common cross-linker glutaraldehyde was compared with a hyperbranched polyglycerol (PG) of around 10 kDa. The latter is oxidized with periodate to generate aldehyde groups that can covalently react with the amines of the surface and the amino groups from the protein via a reductive amination process. The amount of bound protein was quantified via aromatic amino acid analysis (AAAA). This work shows that oxidized polyglycerol can be used as an alternative to glutaraldehyde. With polyglycerol, more of the model protein bovine serum albumin (BSA) could be attached to the surface under the same conditions, and lower nonspecific binding (NSB) was observed. As a proof of concept, IgG was extracted with protein A from crude human plasma. The purity of the product was examined by SDS-PAGE. A binding capacity of 1.8 mg IgG per g of corundum powder was achieved. The advantages of corundum are the very low price, extremely high physical and chemical stability, pressure resistance, favorable binding kinetics, and flexible application.…
Autor*innen: | Jule L. Völzke, Parya Hodjat Shamami, Kornelia GawlitzaORCiD, Ines FeldmannORCiD, Annett Zimathies, Michael G. WellerORCiD |
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Dokumenttyp: | Preprint |
Veröffentlichungsform: | Graue Literatur |
Sprache: | Englisch |
Titel des übergeordneten Werkes (Englisch): | Preprints |
Jahr der Erstveröffentlichung: | 2022 |
Organisationseinheit der BAM: | 1 Analytische Chemie; Referenzmaterialien |
1 Analytische Chemie; Referenzmaterialien / 1.5 Proteinanalytik | |
1 Analytische Chemie; Referenzmaterialien / 1.9 Chemische und optische Sensorik | |
4 Material und Umwelt | |
4 Material und Umwelt / 4.2 Material-Mikrobiom Wechselwirkungen | |
6 Materialchemie | |
6 Materialchemie / 6.3 Strukturanalytik | |
Veröffentlichende Institution: | Bundesanstalt für Materialforschung und -prüfung (BAM) |
Verlag: | MDPI |
Verlagsort: | Basel |
Aufsatznummer: | 2022080004 |
Erste Seite: | 1 |
Letzte Seite: | 13 |
DDC-Klassifikation: | Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Ingenieurwissenschaften und zugeordnete Tätigkeiten |
Freie Schlagwörter: | Affinity chromatography; Antibodies; Aromatic amino acid analysis aaaa; Bioconjugation; Bioseparation; Carrier; Glutaraldehyde; Hyperbranched polymer; Igg; Immunoglobulins; Immunoprecipitation; Linker; Periodate oxidation; Polyglycerol; Protein; Purification; Reductive amination; Self-assembled monolayer (SAM); Solid phase |
Themenfelder/Aktivitätsfelder der BAM: | Material |
Material / Materialdesign | |
DOI: | 10.20944/preprints202208.0004.v1 |
URN: | urn:nbn:de:kobv:b43-555142 |
ISSN: | 2310-287X |
Zugehöriger Identifikator: | https://nbn-resolving.org/urn:nbn:de:kobv:b43-559342 |
Verfügbarkeit des Dokuments: | Datei für die Öffentlichkeit verfügbar ("Open Access") |
Lizenz (Deutsch): | Creative Commons - CC BY - Namensnennung 4.0 International |
Datum der Freischaltung: | 17.08.2022 |
Referierte Publikation: | Nein |
Schriftenreihen ohne Nummerierung: | Preprints der BAM |