Reaktionsgleichung
[CH3:1][C:2]([CH2:4][CH2:5][NH+:6]1[CH2:12][CH2:11][CH2:10][CH2:9][CH2:8][CH2:7]1)=O.[Cl-].C1C([C@@H](O)[C@H](N[C:25]([CH:27](Cl)Cl)=[O:26])CO)=CC=C([N+]([O-])=O)C=1.CC(S[C@@H]1[O:43][C@H](CO)[C@H](O)[C@H](O)[C@H]1O)C>>[CH:9]1[CH:10]=[CH:11][C:12]2[NH:6][CH:5]=[C:4]([CH2:2][CH2:1][CH2:27][C:25]([OH:43])=[O:26])[C:7]=2[CH:8]=1Komponenten
Versuchsvorschrift
The expression of the TG-1 F cells on pMORPHx9-FS-coded Fab fragments in E. coli was performed in shaking bottle cultures with 0.75 l of 2×TY medium and 34 μg/ml of chloramphenicol. After induction with 0.75 mmol of IPTG, the cells were cultivated for 16 hours at 30° C. As an alternative, Fab clones, which had been obtained from the second maturation pool 2, were induced with 0.1 mmol of IPTG and then cultivated at 22° C. Periplasmatic extracts from cell pellets were produced by osmotic shock, and the Fab fragments were isolated by Strep-Tactin® chromatography (IBA, Göttingen, Germany). The apparent molecular weights were determined by size-exclusion chromatography (SEC) with calibrating standards. The concentrations were determined by UV spectrometry.
Aufarbeitung
- customAs an alternative, Fab clones, which had been obtained from the second maturation pool 2
- customcultivated at 22° C
- customPeriplasmatic extracts from cell pellets were produced by osmotic shock
Quellen und Nachweise
- ORD-Eintrag
- ord-bf7b5b23e59047e092d8da6bc3434ef1
- Quelldatensatz
- uspto-grants-2010_08
- Patentnummer
- US07785591B2
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