C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/60 (2006.01) C07K 16/40 (2006.01) C12N 9/88 (2006.01) C12N 15/63 (2006.01) C12N 15/74 (2006.01)
Patent
CA 2192159
The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading ennzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I,II and III are described.
Isolation et séquence de gènes provenant du Flavobacterium heparinum et codant l'héparinase II et l'héparinase III (EC 4.2.2.8), des enzymes qui dégradent l'héparine et le sulfate d'héparane. On décrit également un procédé d'expression et une expression d'héparinase I, II et III utilisant une région de liaison de ribosomes modifiée dérivée d'un promoteur à partir de gènes de glycosaminoglycan lyase de F. Heparinum. On décrit en outre un procédé de purification de protéines comprenant la rupture cellulaire, la chromatographie à échange de cations, la chromatographie par affinité et la chromatographie sur hydroxylapatite, de même que des anticorps contre une fraction de modification après traduction commune aux protéines de Flavobacterium heparinum et un procédé de production d'anticorps spécifiques de ces fractions et des séquences d'acides aminés
Bennett Clark
Blain Francoise
Gu Kangfu
Musil Roy
Su Hongsheng
Biomarin Pharmaceutical Inc.
Ibex Technologies Inc.
Mbm Intellectual Property Law Llp
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