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    Enzymatic cycling assays for homocysteine and cystathionine
    1.
    发明授权
    Enzymatic cycling assays for homocysteine and cystathionine 有权
    同型半胱氨酸和胱硫醚的酶循环试验

    公开(公告)号：US06867014B2

    公开(公告)日：2005-03-15

    申请号：US10651183

    申请日：2003-08-28

    申请人： Glenn Kawasaki ,  Heather Kay Webb ,  Jeffrey Owens ,  Raymond Liedtke ,  Doreen Forest ,  Mark Legaz ,  Sobomabo Lawson

    发明人： Glenn Kawasaki ,  Heather Kay Webb ,  Jeffrey Owens ,  Raymond Liedtke ,  Doreen Forest ,  Mark Legaz ,  Sobomabo Lawson

    IPC分类号： C12Q1/00 ,  C12N9/00 ,  C12N15/52 ,  C12Q1/26 ,  C12Q1/32 ,  C12Q1/527 ,  G01N33/53 ,  G01N33/68 ,  C12Q1/34 ,  C12Q1/48

    CPC分类号： C12Q1/26 ,  C12Q1/32 ,  C12Q1/527 ,  G01N33/6815 ,  Y10S435/968 ,  Y10S435/975

    摘要： The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay. In preferred embodiments, the assays can be conducted in 15 minutes or less, with a minimum of enzyme usage.

    摘要翻译： 本发明提供了用于评估溶液如血液，血液衍生物或尿液中高半胱氨酸和/或胱硫醚的量的酶循环测定。 该测定包括以下步骤：使含有同型半胱氨酸和/或胱硫醚的溶液与CBS或其衍生物，L-丝氨酸和CBL或其衍生物反应一段足以催化周期性 同型半胱氨酸形式转化为胱硫醚，并产生丙酮酸和氨，将胱硫醚转化为同型半胱氨酸; 测定反应混合物中存在的同型半胱氨酸和/或氨的量; 并且基于形成的丙酮酸盐和/或氨的量确定溶液中存在的同型半胱氨酸和/或胱硫醚的量。 还提供用于CBS或其衍生物，CBL或其衍生物的表达载体和分离程序以及用于进行测定的测试试剂盒。 在优选的实施方案中，测定可以在15分钟或更短时间内进行，酶用量最小。

    Enzymatic cycling assays for homocysteine and cystathionine
    2.
    发明授权
    Enzymatic cycling assays for homocysteine and cystathionine 有权
    同型半胱氨酸和胱硫醚的酶循环试验

    公开(公告)号：US06664073B1

    公开(公告)日：2003-12-16

    申请号：US09704036

    申请日：2000-11-01

    申请人： Glenn Kawasaki ,  Heather Kay Webb ,  Jeffrey Owens ,  Raymond Liedtke ,  Doreen Forest ,  Mark Legaz ,  Sobomabo Lawson

    发明人： Glenn Kawasaki ,  Heather Kay Webb ,  Jeffrey Owens ,  Raymond Liedtke ,  Doreen Forest ,  Mark Legaz ,  Sobomabo Lawson

    IPC分类号： C12Q126

    CPC分类号： C12Q1/26 ,  C12Q1/32 ,  C12Q1/527 ,  Y10S435/968 ,  Y10S435/975

    摘要： The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood, derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay. In preferred embodiments, the assays can be conducted in 15 minutes or less, with a minimum of enzyme usage.

    摘要翻译： 本发明提供了用于评估溶液如血液，血液，衍生物或尿液中的同型半胱氨酸和/或胱硫醚的量的酶循环试验。 该测定包括以下步骤：使含有同型半胱氨酸和/或胱硫醚的溶液与CBS或其衍生物，L-丝氨酸和CBL或其衍生物反应一段足以催化周期性 同型半胱氨酸形式转化为胱硫醚，并产生丙酮酸和氨，将胱硫醚转化为同型半胱氨酸; 确定反应混合物中存在的同型半胱氨酸和/或氨的量; 并且基于形成的丙酮酸盐和/或氨的量确定溶液中存在的同型半胱氨酸和/或胱硫醚的量。 还提供用于CBS或其衍生物，CBL或其衍生物的表达载体和分离程序以及用于进行测定的测试试剂盒。 在优选的实施方案中，测定可以在15分钟或更短时间内进行，酶用量最小。