公开(公告)号：US5464747A

公开(公告)日：1995-11-07

申请号：US146633

申请日：1993-10-29

申请人： Scott J. Eisenbeis ,  Mark Krevolin ,  Christopher P. Bryant ,  Sophie J. Boguslawski ,  David J. Ledden ,  Scott Clark

发明人： Scott J. Eisenbeis ,  Mark Krevolin ,  Christopher P. Bryant ,  Sophie J. Boguslawski ,  David J. Ledden ,  Scott Clark

IPC分类号： G01N33/536 ,  C12N9/38 ,  C12N15/09 ,  C12Q1/34 ,  C12R1/19 ,  G01N33/542 ,  G01N33/573 ,  C12N15/56 ,  G01N33/535

CPC分类号： C12Y302/01023 ,  C12N9/2471 ,  G01N33/573 ,  Y10S435/963

摘要： A mutein of an enzyme acceptor polypeptide fragment of .beta.-galactosidase which is resistant to oxidation is provided. The enzyme acceptor fragment has an amino acid other than cysteine located at position 602 of the natural sequence. An especially preferred substitution is serine. Also provided are a method for producing the novel mutein, a reagent composition comprising the novel mutein, and an immunoassay method for determining an analyte in which the novel mutein recombines with an enzyme donor polypeptide fragment to form enzymatically active .beta.-galactosidase.

摘要翻译： 提供耐氧化的β-半乳糖苷酶的酶受体多肽片段的突变蛋白。 酶受体片段具有位于天然序列602位的半胱氨酸以外的氨基酸。 特别优选的取代是丝氨酸。 还提供了用于产生新型突变蛋白的方法，包含新型突变蛋白的试剂组合物和用于测定其中新型突变蛋白与酶供体多肽片段重组以形成酶活性β-半乳糖苷酶的分析物的免疫测定方法。

公开(公告)号：US5492813A

公开(公告)日：1996-02-20

申请号：US146673

申请日：1993-11-01

申请人： Scott J. Eisenbeis ,  Sophie J. Boguslawski ,  Mark Krevolin ,  David J. Ledden

发明人： Scott J. Eisenbeis ,  Sophie J. Boguslawski ,  Mark Krevolin ,  David J. Ledden

IPC分类号： C12N9/38 ,  C12N15/09 ,  C12R1/19 ,  G01N33/535 ,  G01N33/542 ,  C12N15/56

CPC分类号： C12Y302/01023 ,  C12N9/2471 ,  G01N33/535 ,  Y10S435/963

摘要： Muteins of enzyme acceptor polypeptide fragments of .beta.-galactosidase are provided which exhibit substantially increased kinetic complementation activity with no significant loss in stability. A preferred enzyme acceptor fragment has an amino acid other than cysteine located at position 500 of the natural sequence. An especially preferred substitution is serine or valine. Other preferred muteins have an amino acid other than methionine located at position 443, with leucine being especially preferred, or an amino acid other than cysteine at position 76, with leucine being an especially preferred substitution. Also provided are methods for producing the novel muteins, reagent compositions comprising the novel muteins, and immunoassay methods for determining an analyte in which the novel mutein recombines with an enzyme donor polypeptide fragment to form enzymatically active .beta.-galactosidase.

摘要翻译： 提供了β-半乳糖苷酶的酶受体多肽片段的突变蛋白，其表现出显着增加的动力学互补活性，而没有显着的稳定性损失。 优选的酶受体片段具有位于天然序列的位置500处的半胱氨酸以外的氨基酸。 特别优选的取代是丝氨酸或缬氨酸。 其他优选的突变蛋白具有除位于第443位的甲硫氨酸以外的氨基酸，特别优选亮氨酸，或位置76处的半胱氨酸以外的氨基酸，亮氨酸是特别优选的取代基。 还提供了用于产生新型突变蛋白的方法，包含新颖突变蛋白的试剂组合物和用于测定其中新型突变蛋白与酶供体多肽片段重组以形成酶活性β-半乳糖苷酶的分析物的免疫测定方法。

公开(公告)号：US5362625A

公开(公告)日：1994-11-08

申请号：US67342

申请日：1993-05-25

申请人： Mark Krevolin ,  David Kates

发明人： Mark Krevolin ,  David Kates

IPC分类号： C07K14/195 ,  C07K14/00 ,  C07K14/41 ,  C07K19/00 ,  C12N9/38 ,  C12N15/00 ,  C12N15/09 ,  C12Q1/34 ,  G01N33/53 ,  G01N33/58 ,  G01N33/535

CPC分类号： G01N33/581

摘要： The use of omega-acceptor and omega-donor polypeptides (comprising about two-thirds and one-third of the .beta.-galactosidase molecule amino and carboxyl termini, respectively), prepared by recombinant DNA techniques, DNA synthesis, or chemical polypeptide synthesis techniques, which are capable of interacting to form an active enzyme complex having catalytic activity characteristic of .beta.-galactosidase, is described along with improved methods and novel compositions for enzyme complementation assays for qualitative and quantitative determination of a suspected analyte in a sample.

摘要翻译： 通过重组DNA技术，DNA合成或化学多肽合成技术制备的ω-受体和ω-供体多肽（分别包含约三分之二和三分之一的β-半乳糖苷酶分子氨基和羧基末端） 其能够相互作用以形成具有β-半乳糖苷酶特征的催化活性的活性酶复合物，以及改进的方法和用于酶互补测定的新组合物用于定性和定量测定样品中的可疑分析物。

公开(公告)号：US20050227225A1

公开(公告)日：2005-10-13

申请号：US11100373

申请日：2005-04-05

申请人： Mark Krevolin

发明人： Mark Krevolin

IPC分类号： C12N15/09 ,  A61L2/00 ,  A61L12/12 ,  C12Q1/02 ,  C12Q1/04 ,  C12Q1/68 ,  C12Q1/70 ,  G01N33/569 ,  G01N33/68

CPC分类号： A61L2/0082 ,  C12Q1/04 ,  C12Q1/6806 ,  C12Q1/689 ,  G01N33/56911 ,  G01N33/68 ,  C12Q2527/137 ,  C12Q2523/113

摘要： This invention generally relates to stabilized non-blood-based samples in which microbial biomolecules in non-blood-based samples are stable at high temperatures. The stabilized non-blood-based samples include effective amounts of stabilization components to stabilize the microbial biomolecules in the samples. Stabilization components are typically effective at low levels in these stabilized non-blood-based samples. This provides the advantage of minimizing reagent expenses related to the stabilized non-blood-based samples of the invention. The invention also provides methods of stabilizing microbial biomolecules in non-blood-based samples and related kits.

摘要翻译： 本发明一般涉及其中非血液样品中微生物生物分子在高温下稳定的稳定的非血液样品。 稳定的非血液样品包括有效量的稳定组分以稳定样品中的微生物生物分子。 在这些稳定的非血液样品中，稳定组分通常在低水平下是有效的。 这提供了使与本发明的稳定的非血液样品相关的试剂费用最小化的优点。 本发明还提供了在非血液样品和相关试剂盒中稳定微生物生物分子的方法。