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    Glycosylation variants of iduronate 2-sulfatase
    1.
    发明授权
    Glycosylation variants of iduronate 2-sulfatase 有权
    糖尿病糖苷酸2-硫酸酯酶的糖基化变体

    公开(公告)号：US06541254B1

    公开(公告)日：2003-04-01

    申请号：US09685844

    申请日：2000-10-10

    申请人： Peter J. Wilson ,  Charles Phillip Morris ,  Donald Stewart Anson ,  Teresa Occhiodoro ,  Julie Bielicki ,  Peter Roy Clements ,  John Joseph Hopwood

    发明人： Peter J. Wilson ,  Charles Phillip Morris ,  Donald Stewart Anson ,  Teresa Occhiodoro ,  Julie Bielicki ,  Peter Roy Clements ,  John Joseph Hopwood

    IPC分类号： C12N510

    CPC分类号： C12N9/16 ,  A61K38/00

    摘要： The present invention provides a highly glycosylated iduronate-2-sulfatase enzyme comprising an iduronate-2-sulfatase polypeptide with at least 5 kilodalton (kDa) more sugar than iduronate-2-sulfatase purified from a natural source, e.g. human liver. The present invention also provides an enzymatically active polypeptide fragment or variant of such a highly glycosylated iduronate-2-sulfatase. The present invention further provides an isolated nucleic acid encoding iduronate-2-sulfatase, as well as an expression vector, a host cell and a method for producing the present highly glycosylated iduronate-2-sulfatase enzyme. In one embodiment the present invention is directed to a method for producing a glycosylated iduronate-2-sulfatase enzyme which comprises culturing a host cell containing a nucleic acid encoding an enzymatically active iduronate-2-sulfatase polypeptide wherein the host cell glycosylates the polypeptide to a greater degree than a native iduronate-2-sulfatase polypeptide expressed by a natural human liver cell.

    摘要翻译： 本发明提供了高度糖基化的糖尿病2-硫酸酯酶，其包含与天然来源例如纯化的伊曲膦酸酯-2-硫酸酯酶相比，具有至少5千道尔顿（kDa）多糖的伊万二酸-2-硫酸酯酶多肽。 人肝。 本发明还提供了这样一种高度糖基化的阿魏酸-2-硫酸酯酶的酶活性多肽片段或变体。 本发明还提供了编码艾杜糖醛酸-2-硫酸酯酶的分离的核酸，以及表达载体，宿主细胞和用于制备本发明的高度糖基化的阿拉伯糖酸-2-硫酸酯酶的方法。 在一个实施方案中，本发明涉及一种生产糖基化的阿魏酸-2-硫酸酯酶的方法，其包括培养含有编码酶活性的阿魏酸-2-硫酸酯酶多肽的核酸的宿主细胞，其中宿主细胞将多肽糖基化成 比由天然人肝细胞表达的天然异二烯酸2-硫酸酯酶多肽更大程度。

    Amplification and detection process
    5.
    发明授权
    Amplification and detection process 失效
    扩增和检测过程

    公开(公告)号：US5849544A

    公开(公告)日：1998-12-15

    申请号：US374764

    申请日：1995-01-24

    申请人： Raymond John Harris ,  Charles Phillip Morris

    发明人： Raymond John Harris ,  Charles Phillip Morris

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6834 ,  C12Q1/6816 ,  C12Q1/6844 ,  C12Q1/686

    摘要： This method for detecting a target nucleic acid sequence involves amplification and detection in the same vessel and comprises: (a) amplification of the target nucleic sequence in a vessel which is provided with a solid phase capture probe comprising a nucleic acid sequence capable of hybridizing to at least a portion of said amplified target nucleic acid sequence, said capture probe being incapable of participating or not participating in standard nucleic acid sequence amplification processes, (b) bringing a sample suspected of comprising said target nucleic acid sequence into contact with said capture probe under conditions which allow said amplified target nucleic acid sequence to be bound by said capture probe, and (c) detecting the presence of bound target nucleic acid sequence. In a further aspect, the present invention provides an assay system or kit, for detecting a target nucleic acid sequence in a sample suspected of comprising said target nucleic acid sequence, comprising: (a) a capture probe comprising a nucleic acid sequence capable of hydridizing to at least a portion of said amplified target nucleic acid sequence, said capture probe being immobilized on a solid phase support which forms a part of or is insertable into a container for the sample, and said capture probe being incapable of participating in standard nucleic acid sequence amplification processes, (b) reagents for amplification of said target nucleic acid sequence, and (c) means for detecting said target nucleic acid sequence, when bound by said capture probe.

    摘要翻译： PCT No.PCT / AU93 / 00379 Sec。 371日期1995年1月24日 102（e）1995年1月24日PCT PCT 1993年7月26日PCT公布。 公开号WO94 / 02634 日本1994年2月3日该方法用于检测靶核酸序列涉及在同一容器中的扩增和检测，包括：（a）在提供有包含核酸的固相捕获探针的容器中扩增靶核酸序列 能够与所述扩增的靶核酸序列的至少一部分杂交的序列，所述捕获探针不能参与或不参与标准核酸序列扩增过程，（b）将疑似含有所述靶核酸序列的样品置于 在允许所述扩增的靶核酸序列被所述捕获探针结合的条件下与所述捕获探针接触，和（c）检测结合的靶核酸序列的存在。 在另一方面，本发明提供了一种用于检测疑似含有所述靶核酸序列的样品中的靶核酸序列的测定系统或试剂盒，其包含：（a）包含能够进行杂交的核酸序列的捕获探针 至少部分所述扩增的靶核酸序列，所述捕获探针固定在固相载体上，所述固相载体形成样品的一部分或可插入样品的容器中，并且所述捕获探针不能参与标准核酸 序列扩增方法，（b）用于扩增所述靶核酸序列的试剂，和（c）用所述捕获探针结合时检测所述靶核酸序列的方法。