Reaktionsgleichung
[NH2:1][CH:2]([CH2:4][C:5]1[CH:10]=[CH:9][CH:8]=[CH:7][CH:6]=1)[CH3:3].P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.N(S)C(CC1C=CC=CC=1)C.C1C(CN2C(=O)C=CC2=O)CCC(C(ON2C(=O)CCC2=O)=O)C1.Cl>C(=O)([O-])[O-].[K+].[K+].CO>[NH2:1][CH:2]([CH2:4][C:5]1[CH:10]=[CH:9][CH:8]=[CH:7][CH:6]=1)[CH3:3]Komponenten
Versuchsvorschrift
Amphetamine-ATP (0.095 g, 3×10-4 mol, Example 12) was dissolved in 50 ml of 0.12M potassium carbonate in 80% methanol/20% water. After 5 min at room temperature the thiol concentration was determined by reaction with DTNB to be 25 mM. The amphetamine thiol (7.6 ml, 1.9×10-4 mol) was added with stirring to 20.3 ml (0.4 g, 5.9×10-6 mol) of SMCC-BSA, 10/1. The pH of the solution was adjusted to 7 with 1N hydrochloric acid. The container was purged with argon and the solution was stirred at room temperature for 2 hr. The conjugate solution was then dialyzed with a PYROSTART ultrafiltration module against 10 mM morpholineethanesulfonic acid, pH 5.77 and 110 ml of 3.57 mg/ml BSA-amphetamine was collected. The protein solution was frozen at -70° C.
Aufarbeitung
- concentrationAfter 5 min at room temperature the thiol concentration
- customwas determined by reaction with DTNB
- customThe container was purged with argon
- filtrationThe conjugate solution was then dialyzed with a PYROSTART ultrafiltration module against 10 mM morpholineethanesulfonic acid, pH 5.77 and 110 ml of 3.57 mg/ml BSA-amphetamine
- customwas collected
- temperatureThe protein solution was frozen at -70° C.
Quellen und Nachweise
- ORD-Eintrag
- ord-763baf9c6aec4270b6b858c31a580c6a
- Quelldatensatz
- uspto-grants-1996_06
- Patentnummer
- US05525524
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