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    Immunodiagnostic assays using reducing agents
    1.
    发明申请
    Immunodiagnostic assays using reducing agents 有权
    使用还原剂进行免疫诊断测定

    公开(公告)号：US20060263854A1

    公开(公告)日：2006-11-23

    申请号：US11497259

    申请日：2006-08-02

    申请人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

    发明人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

    IPC分类号： C12Q1/68 ,  C12P21/06

    CPC分类号： C07K14/005 ,  C12N2770/24222 ,  C12Q1/707 ,  G01N33/54306 ,  G01N33/54393 ,  G01N33/576 ,  G01N33/5767

    摘要： The present invention relates to a solid phase immunoassay comprising on said solid phase an antigen in the presence of a reducing agent. The present invention also relates to a method for purifying a cysteine containing recombinantly expressed protein comprising at least 2, preferably 3 or 4 and even more preferably all of the following steps: (a) sulphonation of a lysate from recombinant host cells or lysis of recombinant host cells in the presence of guanidinium chloride followed by a subsequent sulphonation of the cell lysate, (b) treatment with a zwitterionic detergent, preferably after removal of the cell debris, (c) purification of the sulphonated version of the recombinant protein or purification of the sulphonated version of the recombinant protein with subsequent removal of the zwitterionic detergent, with said purification being preferably chromatography, more preferably a Ni-IMAC chromatography with said recombinant protein being a His-tagged recombinant protein, (d) desulphonation of the sulphonated version of the recombinant protein, preferably with a molar excess of DTT, (e) storage in the presence of a molar excess of DTT. The present invention also relates to novel HCV NS3 sequences as depicted in FIGS. 1-8.

    摘要翻译： 本发明涉及一种在还原剂存在下在所述固相上含有抗原的固相免疫测定法。 本发明还涉及一种纯化包含重组表达蛋白质的半胱氨酸的方法，其包含至少2个，优选3个或4个，甚至更优选全部以下步骤：（a）来自重组宿主细胞的裂解物的磺化或重组体的裂解 在氯化胍的存在下接受宿主细胞，随后细胞裂解物的磺化，（b）用两性离子洗涤剂处理，优选在去除细胞碎片后，（c）纯化重组蛋白的磺化版本或纯化 所述重组蛋白的磺化版本随后除去两性离子洗涤剂，所述纯化优选为色谱法，更优选为所述重组蛋白为His标记的重组蛋白的Ni-IMAC色谱，（d）磺化版本的 重组蛋白质，优选摩尔过量的DTT，（e）在mo存在下储存 少量DTT。 本发明还涉及如图1和2所示的新型HCV NS3序列。 1 - 8。

    Immunodiagnostic assays using reducing agents
    2.
    发明授权
    Immunodiagnostic assays using reducing agents 有权
    使用还原剂进行免疫诊断测定

    公开(公告)号：US07935490B2

    公开(公告)日：2011-05-03

    申请号：US11497259

    申请日：2006-08-02

    申请人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

    发明人： Geert Maertens ,  Joost Louwagie ,  Alfons Bosman ,  Erwin Sablon ,  Maan Zrein

    IPC分类号： C12Q1/70 ,  G01N33/543 ,  G01N33/53

    CPC分类号： C07K14/005 ,  C12N2770/24222 ,  C12Q1/707 ,  G01N33/54306 ,  G01N33/54393 ,  G01N33/576 ,  G01N33/5767

    摘要： The present invention relates to a solid phase immunoassay comprising on said solid phase an antigen in the presence of a reducing agent. The present invention also relates to a method for purifying a cysteine containing recombinantly expressed protein comprising at least 2, preferably 3 or 4 and even more preferably all of the following steps: (a) sulphonation of a lysate from recombinant host cells or lysis of recombinant host cells in the presence of guanidinium chloride followed by a subsequent sulphonation of the cell lysate, (b) treatment with a zwitterionic detergent, preferably after removal of the cell debris, (c) purification of the sulphonated version of the recombinant protein or purification of the sulphonated version of the recombinant protein with subsequent removal of the zwitterionic detergent, with said purification being preferably chromatography, more preferably a Ni-IMAC chromatography with said recombinant protein being a His-tagged recombinant protein, (d) desulphonation of the sulphonated version of the recombinant protein, preferably with a molar excess of DTT, (e) storage in the presence of a molar excess of DTT. The present invention also relates to novel HCV NS3 sequences as depicted in FIGS. 1-8.

    摘要翻译： 本发明涉及一种在还原剂存在下在所述固相上含有抗原的固相免疫测定法。 本发明还涉及一种纯化包含重组表达蛋白质的半胱氨酸的方法，其包含至少2个，优选3个或4个，甚至更优选全部以下步骤：（a）来自重组宿主细胞的裂解物的磺化或重组体的裂解 在氯化胍的存在下接受宿主细胞，随后细胞裂解物的磺化，（b）用两性离子洗涤剂处理，优选在去除细胞碎片后，（c）纯化重组蛋白的磺化版本或纯化 所述重组蛋白的磺化版本随后除去两性离子洗涤剂，所述纯化优选为色谱法，更优选为所述重组蛋白为His标记的重组蛋白的Ni-IMAC色谱，（d）磺化版本的 重组蛋白质，优选摩尔过量的DTT，（e）在mo存在下储存 少量DTT。 本发明还涉及如图1和2所示的新型HCV NS3序列。 1-8。