公开(公告)号：US5962289A

公开(公告)日：1999-10-05

申请号：US788622

申请日：1997-01-24

申请人： Douglas G. Kilburn ,  Robert C. Miller ,  Richard A.J. Warren ,  Neil R. Gilkes

发明人： Douglas G. Kilburn ,  Robert C. Miller ,  Richard A.J. Warren ,  Neil R. Gilkes

IPC分类号： G01N33/573 ,  A61K39/39 ,  C07K14/55 ,  C07K17/12 ,  C07K19/00 ,  C08B15/00 ,  C08B37/08 ,  C08H8/00 ,  C12N9/16 ,  C12N9/38 ,  C12N9/42 ,  C12N11/12 ,  C12N15/09 ,  C12P21/02 ,  C12Q1/56 ,  C12R1/19 ,  D06P3/60 ,  D21H17/00 ,  G01N33/531 ,  C12N15/00 ,  C12P21/00

CPC分类号： C08H8/00 ,  A61K39/39 ,  C07K14/55 ,  C08B15/00 ,  C08B37/003 ,  C12N11/12 ,  C12N9/16 ,  C12N9/2437 ,  C12N9/2468 ,  C12Q1/56 ,  C12Y302/01004 ,  C12Y302/01074 ,  C12Y302/01091 ,  D21H17/005 ,  G01N33/531 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/034 ,  C07K2319/33 ,  C07K2319/50 ,  C07K2319/75 ,  G01N2333/924 ,  G01N2400/26 ,  Y10S435/803 ,  Y10S530/808 ,  Y10S530/814

摘要： Fusion proteins or conjugates are provided containing an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that binds to a .beta.-1,4-glycan matrix such as cellulose. The substrate binding region is essentially without polysaccharidase activity. In the fusion protein, the substrate binding region is fused or chemically linked to a polypeptide such as an enzyme, a hormone, an immunoglobulin or a protein dye. By contacting the fusion protein with a .beta.-1,4-glycan matrix, the substrate binding region binds to the matrix to immobilize the polypeptide on the matrix. The polypeptide or fusion protein can be removed from the matrix with a protease acting on a protease recognition sequence or with a solution having a low ionic strength or high pH. In the conjugate, the substrate binding region is joined such as by covalent bonding to a non-protein chemical moiety such as a dye, chromophore, fluorescor, radionuclide or enzyme co-factor. By contacting the conjugate with a .beta.-1,4-glycan matrix, the substrate binding region binds to the matrix to immobilize the chemical moiety on the matrix. The conjugate or chemical moiety can be removed with a protease acting on a protease recognition sequence or with a solution having a low ionic strength or high pH.

摘要翻译： 提供融合蛋白或缀合物，其含有具有多糖酶的底物结合区的氨基酸序列，例如结合β-1,4-聚糖基质如纤维素的纤维素酶。 底物结合区基本上不含多糖酶活性。 在融合蛋白中，底物结合区域与诸如酶，激素，免疫球蛋白或蛋白质染料的多肽融合或化学连接。 通过使融合蛋白与β-1,4-聚糖基质接触，底物结合区结合基质将多肽固定在基质上。 多肽或融合蛋白可以用作用于蛋白酶识别序列的蛋白质或具有低离子强度或高pH的溶液从基质中除去。 在缀合物中，底物结合区域例如通过共价键连接到非蛋白质化学部分如染料，发色团，荧光，放射性核素或酶辅因子上。 通过使该缀合物与β-1,4-聚糖基质接触，底物结合区域与基质结合以将化学部分固定在基质上。 缀合物或化学部分可以用作用于蛋白酶识别序列的蛋白酶或具有低离子强度或高pH的溶液来除去。

公开(公告)号：US5874308A

公开(公告)日：1999-02-23

申请号：US585585

申请日：1996-01-16

申请人： Douglas G. Kilburn ,  Keith R. Humphries ,  James G. Doheny ,  Eric Jervis ,  Judie Alimonti

发明人： Douglas G. Kilburn ,  Keith R. Humphries ,  James G. Doheny ,  Eric Jervis ,  Judie Alimonti

IPC分类号： C12N15/09 ,  C07K14/475 ,  C07K14/71 ,  C07K14/715 ,  C07K17/12 ,  C12N5/02 ,  C12N5/0789 ,  C12N9/42 ,  C12R1/01 ,  C07K14/00

CPC分类号： C12N9/2437 ,  C07K14/475 ,  C07K14/71 ,  C12N5/0647 ,  C12Y302/01004 ,  C07K2319/00 ,  C12N2501/125 ,  C12N2501/70

摘要： Methods and compositions are provided for in vitro expansion of growth factor dependent cells. Expansion is effected through the use of growth factor conjugates that include a growth factor such as a steel factor and a polysaccharidase substrate binding region. The conjugates are immobilized by binding of the substrate binding region to a substrate of the polysaccharidase in a growth chamber for the cells.

摘要翻译： 提供了用于体外扩增生长因子依赖性细胞的方法和组合物。 通过使用包括生长因子如钢因子和多糖酶底物结合区的生长因子共轭物来实现扩增。 通过在细胞的生长室中将底物结合区与多糖酶的底物结合来将缀合物固定化。

公开(公告)号：US5624537A

公开(公告)日：1997-04-29

申请号：US309338

申请日：1994-09-20

申请人： Robin F. B. Turner ,  Douglas G. Kilburn ,  Michael R. Phelps

发明人： Robin F. B. Turner ,  Douglas G. Kilburn ,  Michael R. Phelps

IPC分类号： C12Q1/00 ,  G01N27/26

CPC分类号： C12Q1/002 ,  Y10S435/817

摘要： A regenerable biosensor probe adapted for positioning in a bioreactor comprises a selectively permeable interface membrane, a porous protein-receiving matrix adjacent to the interface membrane, an indicating electrode, an inlet conduit through which fresh protein conjugate may flow to the protein-receiving matrix, and an outlet conduit through which spent protein conjugate may be removed from the protein-receiving matrix. A selectively permeable interface membrane, which may be used in a biosensor system to separate biochemical, optical or other processes from an analyte matrix comprises a supporting mesh, a perfluorosulfonic acid polymer impregnated substrate and a homogenous film of perfluorosulfonic acid polymer.A method of preparing this interface membrane comprises fixing a substrate on to a supporting mesh to form a substrate membrane, casting a perfluorosulfonic and polymer on the substrate membrane and curing the product to so formed.

摘要翻译： 适于定位在生物反应器中的可再生生物传感器探针包括选择性渗透界面膜，与界面膜相邻的多孔蛋白质接收基质，指示电极，新鲜蛋白质缀合物可以通过该入口导管流入蛋白质接受基质， 以及可以从蛋白质接收基质中去除废蛋白质缀合物的出口导管。 可用于生物传感器系统以将生物化学，光学或其它方法与分析物基质分离的选择性可渗透界面膜包括支撑网，全氟​​磺酸聚合物浸渍底物和全氟磺酸聚合物的均匀薄膜。 制备该界面膜的方法包括将基底固定在支撑网上以形成基底膜，在基底膜上浇铸全氟磺化物和聚合物并固化所形成的产品。

公开(公告)号：US5340731A

公开(公告)日：1994-08-23

申请号：US865095

申请日：1992-04-08

申请人： Douglas G. Kilburn ,  Robert C. Miller ,  Neil Gilkes ,  R. Antony J. Warren

发明人： Douglas G. Kilburn ,  Robert C. Miller ,  Neil Gilkes ,  R. Antony J. Warren

IPC分类号： G01N33/573 ,  A61K39/39 ,  C07K14/55 ,  C07K17/12 ,  C07K19/00 ,  C08B15/00 ,  C08B37/08 ,  C08H8/00 ,  C12N9/16 ,  C12N9/38 ,  C12N9/42 ,  C12N11/12 ,  C12N15/09 ,  C12P21/02 ,  C12Q1/56 ,  C12R1/19 ,  D06P3/60 ,  D21H17/00 ,  G01N33/531 ,  C12N15/00 ,  C12P21/00

CPC分类号： C08H8/00 ,  A61K39/39 ,  C07K14/55 ,  C08B15/00 ,  C08B37/003 ,  C12N11/12 ,  C12N9/16 ,  C12N9/2437 ,  C12N9/2468 ,  C12Q1/56 ,  C12Y302/01004 ,  C12Y302/01074 ,  C12Y302/01091 ,  D21H17/005 ,  G01N33/531 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/034 ,  C07K2319/33 ,  C07K2319/50 ,  C07K2319/75 ,  G01N2333/924 ,  G01N2400/26 ,  Y10S435/803 ,  Y10S530/808 ,  Y10S530/814

摘要： A fusion protein that can function as a removable label is prepared containing a polypetide such as an enzyme and an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that has essentially no polysaccharidase activity. By contacting the fusion protein with a .beta.-1,4 glycan matrix such as cellulose, the substrate binding region binds to the matrix to immobilize the polypeptide. The polypetide or fusion protein can be removed from the matrix with a protease capable of cleaving a specific protease cleavage site, or with a solution having a low ionic strength or a high pH. The fusion protein can be prepared by recombinant DNA technology.

摘要翻译： 制备可用作可去除标记的融合蛋白，其含有多肽如酶和具有基本上不含多糖酶活性的纤维素酶的多糖酶的底物结合区的氨基酸序列。 通过使融合蛋白与β-1,4-聚糖基质例如纤维素接触，底物结合区结合基质以固定多肽。 可以用能够切割特异性蛋白酶切割位点的蛋白质或用离子强度低或高pH值的溶液从基质中除去多肽或融合蛋白。 融合蛋白可以通过重组DNA技术制备。