Reaktion #593599

ord-bf7b5b23e59047e092d8da6bc3434ef1

Reaktionsgleichung

CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
CC(=O)CC[NH+]1CCCCCC1.[Cl-]
TG-1
O=C(N[C@H](CO)[C@H](O)c1ccc([N+](=O)[O-])cc1)C(Cl)Cl
chloramphenicol
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
→
O=C(O)CCCc1c[nH]c2ccccc12
IBA

Edukte

CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
CC(=O)CC[NH+]1CCCCCC1.[Cl-]
TG-1
—
2
O=C(N[C@H](CO)[C@H](O)c1ccc([N+](=O)[O-])cc1)C(Cl)Cl
chloramphenicol
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG

Reagenzien

Keine

Reaktionsbedingungen

Detaillierte Bedingungen
See reaction.notes.procedure_details.

Aufarbeitung

  1. 1
    SonstigeAs an alternative, Fab clones, which had been obtained from the second maturation pool 2
  2. 2
    Sonstigecultivated at 22° C
  3. 3
    SonstigePeriplasmatic extracts from cell pellets were produced by osmotic shock

Vorschrift

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.

Quelle

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07785591B2uspto-grants-2010_08