Ingenieurwissenschaften und zugeordnete Tätigkeiten
Filtern
Erscheinungsjahr
- 2019 (3) (entfernen)
Dokumenttyp
- Zeitschriftenartikel (3) (entfernen)
Schlagworte
- IgG (2)
- Additive Fertigung (1)
- Affinity chromatography (1)
- Affinitätschromatographie (1)
- Affinitätsextraktion (1)
- Antibody coating (1)
- Antikörper (1)
- Bioconjugation (1)
- Biosensor (1)
- Borosilikatglas (1)
- Combinatorial chemistry (1)
- Combinatorial polymer libraries (1)
- Conjugates (1)
- Druckstabilität (1)
- Drug (1)
- ESI MS (1)
- FPLC (1)
- Glasmonolith (1)
- HPLC (1)
- Immunglobulin (1)
- Immunoaffinity extraction (1)
- Immunoassay (1)
- Immunocapture (1)
- Immunoglobulins (1)
- Immunoprecipitation (1)
- Immunosensor (1)
- MALDI-TOF (1)
- Mass spectrometry (1)
- Oriented immobilization (1)
- PEG (1)
- Polyethylene glycol (1)
- Precision polymer sequencing (1)
- Protein A (1)
- Protein G (1)
- Proximity-enhanced reaction (1)
- Pseudo peptides (1)
- Regeneration (1)
- Sequence-defined oligomer (1)
- Sequencing (1)
- Solubilizer (1)
- Titan-Halterung (1)
- biokompatibel (1)
- gesintert (1)
Organisationseinheit der BAM
Therapeutische Antikörper sind innerhalb weniger Jahre zur wichtigsten pharmazeutischen Produktklasse aufgestiegen. Für 2023 werden weltweite Umsätze von über 200 Milliarden USD erwartet. Auch diagnostische Antikörper sind mittlerweile unverzichtbare Produkte, auf deren Basis zahllose Immunoassays und andere Schnelltests entwickelt wurden. Neben der bereits sehr aufwendigen Herstellung von Antikörpern ist deren Aufreinigung aus komplexen Zellkulturmedien oder Blutseren und -plasmen zu einem Engpass in der Produktion und Nutzung dieser komplexen Proteine geworden. Schnelle und einfache Reinigungsmethoden für Antikörper sind daher sehr gefragt.
Preactivation Crosslinking - An Efficient Method for the Oriented Immobilization of Antibodies
(2019)
Crosslinking of proteins for their irreversible immobilization on surfaces is a proven and popular method. However, many protocols lead to random orientation and the formation of undefined or even inactive by-products. Most concepts to obtain a more targeted conjugation or immobilization requires the recombinant modification of at least one binding partner, which is often impractical or prohibitively expensive. Here a novel method is presented, which is based on the chemical preactivation of Protein A or G with selected conventional crosslinkers. In a second step, the antibody is added, which is subsequently crosslinked in the Fc part. This leads to an oriented and covalent immobilization of the immunoglobulin with a very high yield. Protocols for Protein A and Protein G with murine and human IgG are presented. This method may be useful for the preparation of columns for affinity chromatography, immunoprecipitation, antibodies conjugated to magnetic particles, permanent and oriented immobilization of antibodies in biosensor systems, microarrays, microtitration plates or any other system, where the loss of antibodies needs to be avoided, and maximum binding capacity is desired. This method is directly applicable even to antibodies in crude cell culture supernatants, raw sera or protein-stabilized antibody preparations without any purification nor enrichment of the IgG. This new method delivered much higher signals as a traditional method and, hence, seems to be preferable in many applications.
Functional sequences of precision polymers based on thiolactone/Michael chemistry are identified from a large one-bead one-compound library. Single-bead readout by MALDI-TOF MS/MS identifies sequences that host m-THPC that is a second Generation photo-sensitizer drug. The corresponding Tla/Michael-PEG conjugates make m-THPC available in solution and drug payload as well as drug release kinetics can be fine-tuned by the precision segment.