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    CORE-GLYCOSYLATED HCV ENVELOPE PROTEINS
    6.
    发明申请
    CORE-GLYCOSYLATED HCV ENVELOPE PROTEINS 审中-公开
    核心糖蛋白HCV包膜蛋白

    公开(公告)号：WO2002086101A2

    公开(公告)日：2002-10-31

    申请号：PCT/BE2002/000064

    申请日：2002-04-24

    Applicant: INNOGENETICS N.V. ,  DEPLA, Erik ,  BOSMAN, Alfons ,  DESCHAMPS, Geert ,  SABLON, Erwin ,  SUCKOW, Manfred ,  SAMSON, Isabelle ,  VERHEYDEN, Gert

    Inventor： DEPLA, Erik ,  BOSMAN, Alfons ,  DESCHAMPS, Geert ,  SABLON, Erwin ,  SUCKOW, Manfred ,  SAMSON, Isabelle ,  VERHEYDEN, Gert

    IPC: C12N1/16

    CPC classification number: C07K14/005 ,  A61K38/00 ,  A61K39/00 ,  A61K2039/5258 ,  C07K2319/02 ,  C07K2319/50 ,  C12N7/00 ,  C12N2770/24222 ,  C12N2770/24223

    Abstract: The current invention relates to HCV envelope proteins or parts thereof which are the product of expression in eukaryotic cells. More particularly said HCV envelope proteins are characterized in that on average up to 80 % of their N-glycosylation sites are core-glycosylated. Of these N-glycosylated sites more than 70 % are glycosylated with an oligomannose containing 8 to 10 mannoses. Furthermore, the ratio of the oligomannoses with structure Man(7)-GlcNAc(2) over the oligomannose with structure Man(8)-GlcNAc(2) is less than or equal to 0.45. Less than 10 % of the oligomannoses is terminated with an α1,3 linked mannose. The HCV envelope proteins of the invention are particularly suited for diagnostic, prophylactic and therapeutic purposes. A suitable eukaryotic cell for production of the HCV envelope proteins of the invention is a Hansenula cell.

    Abstract translation: 本发明涉及作为在真核细胞中表达的产物的HCV包膜蛋白或其部分。 更具体地说，HCV包膜蛋白的特征在于平均高达80％的N-糖基化位点是核糖基化的。 在这些N-糖基化位点中，超过70％的糖苷化与含有8至10个甘露糖的寡甘露糖糖基化。 此外，结构Man（7）-GlcNAc（2）的寡甘露糖与结构Man（8）-GlcNAc（2）的寡甘露糖的比例小于或等于0.45。 少于10％的寡甘露聚糖用α1,3连接的甘露糖终止。 本发明的HCV包膜蛋白特别适用于诊断，预防和治疗目的。 用于生产本发明的HCV包膜蛋白的合适的真核细胞是Hansenula细胞。

    METHODS FOR COVALENT IMMOBILISATION OF BIOMOLECULES TO A CARRIER BY MEANS OF A HIS-TAG
    8.
    发明申请
    METHODS FOR COVALENT IMMOBILISATION OF BIOMOLECULES TO A CARRIER BY MEANS OF A HIS-TAG 审中-公开
    通过其标签将生物分子对载体进行共价偶联的方法

    公开(公告)号：WO9900670A8

    公开(公告)日：2000-03-02

    申请号：PCT/EP9803883

    申请日：1998-06-25

    Applicant: INNOGENETICS NV ,  BOSMAN ALFONS ,  WIJNENDAELE FRANS VAN ,  DEN BROECK DIRK VAN ,  VOORDE ANDRE VAN DE

    Inventor： BOSMAN ALFONS ,  VAN WIJNENDAELE FRANS ,  VAN DEN BROECK DIRK ,  VAN DE VOORDE ANDRE

    IPC: C07K17/00 ,  C07K17/06 ,  C12N11/00 ,  C12N11/06 ,  G01N33/543 ,  G01N33/547

    CPC classification number: C12N11/00 ,  G01N33/54353 ,  G01N33/54393

    Abstract: The present invention relates to methods for covalent immobilisation of biomolecules to carriers and membranes, wherein the presence of a His-tag is exploited, and wherein the amino acid residues that comprise said His-tag are directly involved in the covalent bond. The present invention also provides several strategies that further augment the probability of covalent immobilisation through said His-tags, such as improving the presentation of said His-tag, choosing the appropriate reaction conditions such as pH, temperature, concentration of reagent and kinetics, increasing contact between His-tag and reactive groups of said carrier or membrane, by for instance the use of IDA or anti-His antibodies or increasing the hydrophobicity of the membrane, or shielding the rest of the biomolecule from reaction by for instance increasing the hydrophobicity of said carrier or membrane or addition of substrate or competitors or blocking otherwise reactive groups, or by choosing chemical reactions that have a high selectivity for histidine residues. A carrier can also be another biomolecule. The present invention thus also relates to a method that allows covalent cross-linking between identical or different biomolecules. When such biomolecules have a natural tendency to interact with each other to form homo- or heterodimers, a strategy of increasing contact between the reactive groups (two His-tags or one His-tag and another reactive group) can be exploited. The present invention also relates to a method of providing a simultaneous and universal system for detection of biomolecules through said His-tag.

    Abstract translation: 本发明涉及用于共价固定生物分子到载体和膜的方法，其中使用His标签的存在，其中包含所述His-标签的氨基酸残基直接参与共价键。 本发明还提供了几种策略，其进一步增加通过所述His标签共价固定的概率，例如改善所述His标签的呈现，选择合适的反应条件，例如pH，温度，试剂浓度和动力学，增加 His标签和所述载体或膜的反应性基团之间的接触，例如使用IDA或抗His抗体或增加膜的疏水性，或屏蔽其余生物分子免受反应，例如增加疏水性 所述载体或膜或添加底物或竞争剂或阻断另外的反应性基团，或通过选择对组氨酸残基具有高选择性的化学反应。 载体也可以是另一种生物分子。 因此，本发明还涉及允许相同或不同生物分子之间共价交联的方法。 当这种生物分子具有彼此相互作用以形成同二聚体或异二聚体的天然趋势时，可以利用增加反应性基团（两个His-标签或一个His-标签和另一个反应性基团）之间接触的策略。 本发明还涉及提供用于通过所述His标签检测生物分子的同时和通用系统的方法。

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