C - Chemistry – Metallurgy – 12 – P
Patent
C - Chemistry, Metallurgy
12
P
C12P 21/04 (2006.01)
Patent
CA 2613471
Cell lines are provided for expressing peptidylglycine alpha-amidating monooxygenase (PAM), or one of its two catalytic domains. High levels of enzyme expression are achieved while utilizing a non-animal source, low protein tissue culture medium. A robust two-step downstream purification results in high enzyme purity. Resulting PAM, or its PHM catalytic domain, is used to catalyze the enzymatic conversion of X-Gly to X-alpha-hydroxy-Gly or X- NH2 (X being a peptide or any chemical compound having a carbonyl group to which a glycine group can be covalently attached). Methods of preparing preferred cell lines are also set forth.
L'invention porte sur des lignées cellulaires qui permettent d'exprimer la peptidylglycine alpha-amidating monooxygénase (PAM) ou l'un de ses deux domaines catalytiques. On obtient des niveaux élevés d'expression enzymatique tout en utilisant un milieu de culture tissulaire à faible teneur en protéines de source non animale. Une purification aval robuste en deux étapes permet d'arriver à une haute pureté enzymatique. On utilise la PAM obtenue, ou son domaine catalytique PHM, pour catalyser la conversion enzymatique de X-Gly en X-alpha-hydroxy-Gly ou en X-NH2(X étant un peptide ou un quelconque composé chimique renfermant un groupe carbonyle auquel un groupe glycine peut être attaché par covalence). L'invention concerne également des procédés de préparation de lignées cellulaires préférées.
Consalvo Angelo P.
Mehta Nozer M.
Miller Duncan A.
Marks & Clerk
Unigene Laboratories Inc.
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