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    Recombinant expression of Rubella E1 envelope protein variants as chaperone fusion-proteins, and their use in the detection of anti-Rubella antibodies
    1.
    发明公开
    Recombinant expression of Rubella E1 envelope protein variants as chaperone fusion-proteins, and their use in the detection of anti-Rubella antibodies 有权
    风疹E1包膜蛋白的重组表达的变体作为伴侣融合蛋白和其用于检测抗风疹抗体的使用

    公开(公告)号：EP1780282A1

    公开(公告)日：2007-05-02

    申请号：EP06022216.3

    申请日：2006-10-24

    申请人： Roche Diagnostics GmbH ,  F.HOFFMANN-LA ROCHE AG

    发明人： Bollhagen, Ralf ,  Engel, Alfred ,  Faatz, Elke ,  Schaarschmidt, Peter ,  Scholz, Christian ,  Upmeier, Barbara ,  Zarnt, Toralf

    IPC分类号： C12N15/40 ,  C12N15/62 ,  C12N5/10 ,  C07K14/19 ,  G01N33/569

    CPC分类号： C07K14/005 ,  A61K39/00 ,  C07K2319/35 ,  C12N2770/36222

    摘要： The present invention relates to a soluble Rubella E1 antigen and variants of this peptide characterized by lacking at the C-terminal end at least the transmembran region and the anchor segment as well as at least the amino acids 143 to 164 and containing at least the region spanning the disulfid-bridges Cys 349 - Cys 352 and Cys 368 - 401 whereas the N-terminus (Cys 349) of this region contains additionally at least 15 amino acids and / or the C-terminus (Cys 401) of this region contains additionally at least 8 amino acids of the adjacent Rubella E1 antigen sequence.
    The region spanning the disulfid-bridges Cys 349 - Cys 352 and Cys 368 - 401 contains at the N-terminus of this region (Cys 349) additionally at least 25, 30, 34 amino acids and / or at the C-terminus (Cys 401) of this region additionally at least 10, 11, 15, 25, 35 amino acids of the adjacent Rubella E1 antigen sequence.
    Furthermore, the invention relates to a recombinant DNA molecule, encoding a Rubella E1 antigen and variants, which are recombinantly expressed as a chaperone fusion-protein, refolded into a soluble and immunoreactive conformation and further used for the serological detection of anti-Rubella antibodies. In addition, the present invention discloses a method for the detection, determination and quantification of anti-Rubella antibodies of IgG and / or IgM subclass in a sample wherein the Rubella E1 antigen is used as a capture reagent and / or binding partner for the antibodies. The invention comprises further a diagnostic test and a reagent kit for the detection of anti-Rubella antibodies, containing at least one antigen of the Rubella E1 antigens.

    摘要翻译： 本发明涉及这种肽通过在C末端的至少跨膜区和锚定区段以及至少氨基酸143-164且含有至少该区域缺乏为特征的可溶性风疹E1抗原和变体 跨越二硫键桥的Cys 349 - 半胱氨酸352和Cys 368-401鉴于N-末端（半胱氨酸349）的此区域包含附加的至少15个氨基酸和/或C末端（半胱氨酸401）的此区域包含附加 邻风疹E1抗原的序列的至少8个氨基酸。 跨越二硫键的Cys 349的区域 - 的Cys 352和Cys 368-401包含该区域的N末端（半胱氨酸349）附加的至少25％，30，34个氨基酸和/或在C-末端（半胱氨酸 401）该区域的附加的至少10，11，15，25，风疹E1抗原的相邻序列的35个氨基酸。 进一步，本发明涉及一种重组DNA分子，编码风疹E1抗原和变体，它们是重组过表达作为伴侣融合蛋白，重折叠为可溶性和免疫反应性构象和进一步用于抗风疹抗体的血清学检测。 此外，本发明盘松动用于检测，确定和worin风疹E1抗原的样品中的IgG和/或IgM亚类的抗风疹抗体的定量的方法被用作捕获试剂和/或结合伴侣的抗体 ， 本发明还包括诊断测试和用于检测抗风疹抗体的试剂盒，含有该风疹E1抗原的至少一种抗原。