公开(公告)号：US6090567A

公开(公告)日：2000-07-18

申请号：US445464

申请日：1995-05-22

申请人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

发明人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

IPC分类号： G01N33/542 ,  C12N9/04 ,  C12N15/09 ,  C12Q1/28 ,  C12Q1/32 ,  C12R1/19 ,  G01N33/535 ,  G01N33/569 ,  G01N33/58 ,  G01N33/78 ,  G01N33/94 ,  G01N33/53

CPC分类号： C12N9/0006 ,  C12Q1/28 ,  G01N33/535 ,  G01N33/56911 ,  G01N33/581 ,  G01N33/78 ,  G01N33/94 ,  G01N2333/904

摘要： Methods for immunoassay of analytes employing mutant glucose-6-phosphate dehydrogenase (G6PDH) enzymes as labels. In particular, the invention relates to the use of conjugates of an analyte or analyte analog and a mutant NAD.sup.+ dependent G6PDH differing from any precursor G6PDH by the deletion, substitution, or insertion, or any combination thereof of at least one amino acid per subunit. The invention also involves the construction of several mutations in precursor glucose-6-phosphate dehydrogenase (G6PDH) enzymes. Typically, the mutations involve deletion or substitution of one or more lysine residues, or introduction of one or more cysteine residues by insertion of cysteine to precursor G6PDH or substitution of precursor G6PDH amino acids residues with cysteine. The present invention also relates to conjugates of the subject enzymes and specific binding pair members, kits useful in performing the methods of the invention, cell lines producing the subject enzymes, DNA sequences encoding the subject enzymes, and vectors containing DNA encoding the subject enzymes and designed to allow a host cell to produce the subject enzymes.

摘要翻译： 使用突变型葡萄糖-6-磷酸脱氢酶（G6PDH）酶作为标记免疫测定分析物的方法。 特别地，本发明涉及通过每个亚基的至少一个氨基酸的缺失，取代或插入或其任何组合，使用不同于任何前体G6PDH的分析物或分析物类似物和突变体NAD +依赖性G6PDH的缀合物的用途。 本发明还涉及前体葡萄糖-6-磷酸脱氢酶（G6PDH）酶的几个突变的构建。 通常，突变涉及一个或多个赖氨酸残基的缺失或取代，或通过将半胱氨酸插入前体G6PDH或用半胱氨酸取代前体G6PDH氨基酸残基引入一个或多个半胱氨酸残基。 本发明还涉及本发明的酶和特异性结合对成员的缀合物，可用于实施本发明方法的试剂盒，产生本发明酶的细胞系，编码本发明酶的DNA序列，和含有编码本发明酶的DNA的载体和 旨在使宿主细胞产生目标酶。

公开(公告)号：US06455288B1

公开(公告)日：2002-09-24

申请号：US08044857

申请日：1993-04-08

申请人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

发明人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

IPC分类号： C12N1552

CPC分类号： C12N9/0006 ,  C12Q1/28 ,  G01N33/535 ,  G01N33/56911 ,  G01N33/581 ,  G01N33/78 ,  G01N33/94 ,  G01N2333/904

摘要： The present invention relates to methods for immunoassay of analytes employing mutant glucose-6-phosphate dehydrogenase (G6PDH) enzymes as labels. In particular, the invention relates-to the use of conjugates of an analyte or analyte analog and a mutant NAD+ dependent G6PDH differing from any precursor G6PDH by the deletion, substitution, or insertion, or any combination thereof of at least one amino acid per subunit. The invention also involves the construction of several mutations in precursor glucose-6-phosphate dehydrogenase (G6PDH) enzymes. Typically, the mutations involve deletion or substitution of one or more lysine residues, or introduction of one or more cysteine residues by insertion of cysteine to precursor G6PDH or substitution of precursor G6PDH amino acids residues with cysteine. The present invention also relates to conjugates of the subject enzymes and specific binding pair members, kits useful in performing the methods of the invention, cell lines producing the subject enzymes, DNA sequences encoding the subject enzymes, and vectors containing DNA encoding the subject enzymes and designed to allow a host cell to produce the subject enzymes.

摘要翻译： 本发明涉及使用突变型葡萄糖-6-磷酸脱氢酶（G6PDH）酶作为标记免疫测定分析物的方法。 特别地，本发明涉及分离物或分析物类似物和与任何前体G6PDH不同的突变体NAD +依赖性G6PDH通过每个亚基的缺失，取代或插入或其任何组合的至少一个氨基酸的缀合物的用途 。 本发明还涉及前体葡萄糖-6-磷酸脱氢酶（G6PDH）酶的几个突变的构建。 通常，突变涉及一个或多个赖氨酸残基的缺失或取代，或通过将半胱氨酸插入前体G6PDH或用半胱氨酸取代前体G6PDH氨基酸残基引入一个或多个半胱氨酸残基。 本发明还涉及本发明的酶和特异性结合对成员的缀合物，可用于实施本发明方法的试剂盒，产生本发明酶的细胞系，编码本发明酶的DNA序列，和含有编码本发明酶的DNA的载体和 旨在使宿主细胞产生目标酶。

公开(公告)号：US6033890A

公开(公告)日：2000-03-07

申请号：US445463

申请日：1995-05-22

申请人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

发明人： Edward Benjamin Jakobovits ,  Joy L. Silen ,  Mark J. Levy ,  Thomas C. Goodman ,  Martin Becker ,  Edwin F. Ullman ,  Robert M. Caldwell ,  Richard R. Bott ,  Christopher Charles Barnett

IPC分类号： G01N33/542 ,  C12N9/04 ,  C12N15/09 ,  C12Q1/28 ,  C12Q1/32 ,  C12R1/19 ,  G01N33/535 ,  G01N33/569 ,  G01N33/58 ,  G01N33/78 ,  G01N33/94

CPC分类号： C12N9/0006 ,  C12Q1/28 ,  G01N33/535 ,  G01N33/56911 ,  G01N33/581 ,  G01N33/78 ,  G01N33/94 ,  G01N2333/904

摘要： The present invention relates to methods for immunoassay of analytes employing mutant glucose-6-phosphate dehydrogenase (G6PDH) enzymes as labels. In particular, the invention relates to the use of conjugates of an analyte or analyte analog and a mutant NAD.sup.+ dependent G6PDH differing from any precursor G6PDH by the deletion, substitution, or insertion, or any combination thereof of at least one amino acid per subunit. The invention also involves the construction of several mutations in precursor glucose-6-phosphate dehydrogenase (G6PDH) enzymes. Typically, the mutations involve deletion or substitution of one or more lysine residues, or introduction of one or more cysteine residues by insertion of cysteine to precursor G6PDH or substitution of precursor G6PDH amino acids residues with cysteine. The present invention also relates to conjugates of the subject enzymes and specific binding pair members, kits useful in performing the methods of the invention, cell lines producing the subject enzymes, DNA sequences encoding the subject enzymes, and vectors containing DNA encoding the subject enzymes and designed to allow a host cell to produce the subject enzymes.

摘要翻译： 本发明涉及使用突变型葡萄糖-6-磷酸脱氢酶（G6PDH）酶作为标记免疫测定分析物的方法。 特别地，本发明涉及通过每个亚基的至少一个氨基酸的缺失，取代或插入或其任何组合，使用不同于任何前体G6PDH的分析物或分析物类似物和突变体NAD +依赖性G6PDH的缀合物的用途。 本发明还涉及前体葡萄糖-6-磷酸脱氢酶（G6PDH）酶的几个突变的构建。 通常，突变涉及一个或多个赖氨酸残基的缺失或取代，或通过将半胱氨酸插入前体G6PDH或用半胱氨酸取代前体G6PDH氨基酸残基引入一个或多个半胱氨酸残基。 本发明还涉及本发明的酶和特异性结合对成员的缀合物，可用于实施本发明方法的试剂盒，产生本发明酶的细胞系，编码本发明酶的DNA序列，和含有编码本发明酶的DNA的载体和 旨在使宿主细胞产生目标酶。