Reaktion #42622

ord-2d2981e0a166410dbf5d3f4dd745d44d

Reaktionsgleichung

CC(=O)O.CC(C)C[C@H](NC(=O)CNC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@H](CO)NC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@H](Cc1cnc[nH]1)NC(=O)[C@@H]1CCC(=O)N1)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N1CCC[C@H]1C(=O)NCC(N)=O.NCC(=O)N[C@@H](CS)C(=O)O

GnRH Gly-Cys

→

CC(=O)O.CC(C)C[C@H](NC(=O)CNC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@H](CO)NC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@H](Cc1cnc[nH]1)NC(=O)[C@@H]1CCC(=O)N1)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N1CCC[C@H]1C(=O)NCC(N)=O

GnRH

Edukte

GnRH Gly-Cys

Reagenzien

Keine

Lösungsmittel

H2O

Reaktionsbedingungen

Detaillierte Bedingungen

See reaction.notes.procedure_details.

Aufarbeitung

1
workup.ADDITIONcontaining an free SH on one end)
2
workup.ADDITIONThe dissolved hapten is added to the activated KLH
3
Sonstigeto react for 2 hours at room temperature
4
Sonstigeovernight
5
SonstigeThe KLH-hapten conjugate is immediately purified
6
TemperaturThe GnRH/KLH conjugate may be frozen

Vorschrift

Six mg of the GnRH-Gly-Cys hapten (containing an free SH on one end) is dissolved in 1 ml of H2O. The dissolved hapten is added to the activated KLH and allowed to react for 2 hours at room temperature and then overnight in the refrigerator. The KLH-hapten conjugate is immediately purified by applying the reaction mixture to a desalting column. (Sephadex G-25). The conjugate protein should come off on the void volume (first peak, fractions 4-6) as measure by absorbance at 280 nm. There should be a drop in absorbance after fraction 6 and rise in absorbance in fractions 7 or 8 as the excess hapten comes off. A small amount of excess hapten does not cause problem in a prime dose but could neutralize antibody in a boost dose if it is present in large excess. The GnRH/KLH conjugate may be frozen or lyophilized or used in the present form.

Quelle

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07731939B2uspto-grants-2010_06