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1. 发明授权

US6048715A Two-phase partition affinity separation system and affinity separated cell-containing composition 失效
标题翻译：两相分离亲和分离系统和亲和力分离含细胞的组合物
公开(公告)号：US6048715A
公开(公告)日：2000-04-11
申请号：US685808
申请日：1996-07-24
申请人： Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
发明人： Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
IPC分类号： A61K39/39 , C07K1/14 , C07K14/55 , C08B15/00 , C08B37/08 , C08H8/00 , C12N9/16 , C12N9/38 , C12N9/42 , C12N11/12 , C12P19/02 , C12Q1/56 , D21H17/00 , G01N33/531 , A23J1/00 , C12N15/00 , C12P21/06
CPC分类号： C08H8/00 , A61K39/39 , C07K1/14 , C07K14/55 , C08B15/00 , C08B37/003 , C12N11/12 , C12N9/16 , C12N9/2437 , C12N9/2445 , C12P19/02 , C12Q1/56 , C12Y302/01004 , C12Y302/01021 , D21H17/005 , G01N33/531 , C07K2319/00 , G01N2333/924 , G01N2400/26 , Y10S435/803 , Y10S435/815 , Y10S530/813 , Y10S530/814
摘要： A two-phase partition system is provided for affinity separation of a composition containing a polysaccharide binding peptide from a mixture such as a fermentation broth. The peptide may be from an enzyme and lacking in polysaccharidase activity such as the binding domain of cellulase that binds to cellulose. The system contains a phase-forming oligosaccharide polymer such as a cellulose derivative to which the peptide binds with a Ka of 10.sup.3 M to 10.sup.7 M, and a phase inducing agent such as a polyethylene glycol polymer, or a salt present at sufficiently high concentration to induce phase separation. If the oligosaccharide polymer is thermoseparating, phase separation can be induced by heating. Using the system involves contacting a composition containing the peptide such as a fusion protein with the system, partitioning the composition into a phase containing the oligosaccharide polymer by binding to the polymer and recovering the polymer containing the bound composition. The peptide or a fusion protein containing the peptide can be contacted with a cell having a carbohydrate residue to which the peptide binds to form a complex, and the complex is separated with the system to produce a bound cell composition. The peptide may be linked through a protease recognition sequence to a macromolecule such as an enzyme, a hormone or an antibody, and the macromolecule can be removed by using a protease to cleave the recognition sequence.
摘要翻译：提供了两相隔离系统，用于从包含发酵液的混合物中亲和分离含有多糖结合肽的组合物。肽可以来自酶并且缺乏多糖酶活性，例如与纤维素结合的纤维素酶的结合结构域。该体系含有形成型寡糖聚合物，例如纤维素衍生物，其肽与103M至107M的Ka结合，以及相诱导剂如聚乙二醇聚合物或以足够高的浓度存在的盐诱导相分离。如果寡糖聚合物是热分离的，则可以通过加热诱导相分离。使用该系统包括使包含诸如融合蛋白的肽的组合物与系统接触，通过与聚合物结合并将含有结合的组合物的聚合物回收到含有寡糖聚合物的相中，将组合物分配。可以将肽或含有该肽的融合蛋白与具有肽结合的碳水化合物残基的细胞接触以形成复合物，并且用体系分离复合物以产生结合的细胞组合物。肽可以通过蛋白酶识别序列连接到大分子如酶，激素或抗体，并且可以通过使用蛋白酶来切割识别序列来除去大分子。

2. 发明授权

US06174700B1 Purification of a polypeptide compound having a polysaccharide binding domain by affinity phase separation 失效
标题翻译：通过亲和相分离纯化具有多糖结合结构域的多肽化合物
公开(公告)号：US06174700B1
公开(公告)日：2001-01-16
申请号：US08505860
申请日：1995-07-24
申请人： Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
发明人： Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
IPC分类号： C12P2106
CPC分类号： A61K39/39 , C07K1/14 , C07K14/55 , C07K2319/00 , C08B15/00 , C08B37/003 , C08H8/00 , C12N9/16 , C12N9/2437 , C12N9/2445 , C12N11/12 , C12P19/02 , C12Q1/56 , C12Y302/01004 , C12Y302/01021 , D21H17/005 , G01N33/531 , G01N2333/924 , G01N2400/26 , Y10S435/803 , Y10S435/815 , Y10S530/813 , Y10S530/814
摘要： A compound having a polysaccharide binding domain such as contained by a cellulose and essentially lacking in polysaccharidase activity is purified from other ingredients in a mixture using an affinity partition system. A mixture containing the compound is contacted with a system containing as a first phase an aqueous solution of oligosaccharide polymer such as cellulose and as a second phase a solution of a polymer such as a poly(ethylene glycol)-poly(propylene glycol) copolymer. The compound petitions into the first phase and binds to the oligosaccharide polymer, preferably with a Ka of 103 to 107, to form a complex. The complex is collected, and the compound is dissociated from the oligosaccharide polymer. The compound may be formed of a non-peptide chemical moiety or a peptide moiety linked to a polypeptide having the polysaccharide binding domain. The compound may also be a fusion polypeptide containing the polysaccharide binding domain linked through a protease recognition sequence to a macromolecule such as an enzyme, a hormone or an antibody. The macromolecule can be removed by using a protease to cleave the recognition sequence. Another partition system contains the oligosaccharide polymer and a phase separation inducing agent such as a sulfate or citrate salt that induces separation to produce different phases.
摘要翻译：具有多糖结合结构域的化合物，例如由纤维素包含并且基本上不含多糖酶活性的化合物，使用亲和分配系统从混合物中的其它成分中纯化。将含有化合物的混合物与含有第一相的系统接触，例如纤维素等低聚糖聚合物的水溶液，作为第二相与聚（乙二醇） - 聚（丙二醇）共聚物等聚合物的溶液接触。化合物进入第一阶段并结合寡糖聚合物，优选用103至107的Ka形成络合物。收集该络合物，并且该化合物与寡糖聚合物解离。化合物可以由非肽化学部分或与具有多糖结合结构域的多肽连接的肽部分形成。化合物还可以是含有通过蛋白酶识别序列连接到大分子如酶，激素或抗体的多糖结合结构域的融合多肽。可以通过使用蛋白酶切割识别序列来除去大分子。另一分隔系统含有寡糖聚合物和相分离诱导剂如硫酸盐或柠檬酸盐，其诱导分离以产生不同相。

3. 发明授权

US5928917A Conjugate of non-protein chemical moiety and polypeptide having cellulose binding region 失效
标题翻译：具有纤维素结合区域的非蛋白质化学部分和多肽的缀合物
公开(公告)号：US5928917A
公开(公告)日：1999-07-27
申请号：US249037
申请日：1994-05-24
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： 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 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。可以用蛋白酶识别序列或具有低离子强度或高pH的溶液从基质中除去多肽或融合蛋白。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。

4. 发明授权

US5821358A Methods and compositions for modification of polysaccharide characteristics 失效
标题翻译：用于修饰多糖特征的方法和组合物
公开(公告)号：US5821358A
公开(公告)日：1998-10-13
申请号：US269614
申请日：1994-06-30
申请人： Neil R. Gilkes , Douglas G. Kilburn , Robert C. Miller, Jr. , Anthony Warren
发明人： Neil R. Gilkes , Douglas G. Kilburn , Robert C. Miller, Jr. , Anthony Warren
IPC分类号： D06M16/00 , C12P19/04
CPC分类号： D06M16/003
摘要： Methods and compositions are provided for the modification of polysaccharide structures using polysaccharidase binding or catalytic domains either alone or in tandem to modify the structure of polysaccharides. These methods and compositions are exemplified by the use of cellulase binding and catalytic domains to polish cotton, and to alter dying characteristics, texture and porosity of cellulose fibers.
摘要翻译：提供了使用多糖酶结合或催化结构域单独或串联修饰多糖结构以修饰多糖结构的方法和组合物。这些方法和组合物的例子是使用纤维素酶结合和催化结构域来抛光棉花，并改变纤维素纤维的染色特性，质地和孔隙率。

5. 发明授权

US5202247A Cellulose binding fusion proteins having a substrate binding region of cellulase 失效
标题翻译：具有纤维素酶底物结合区域的纤维素结合融合蛋白
公开(公告)号：US5202247A
公开(公告)日：1993-04-13
申请号：US603987
申请日：1990-10-25
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： C12P19/14 , A61K39/39 , C07K1/22 , C07K14/00 , C07K14/195 , C07K14/41 , C07K14/52 , C07K14/54 , C07K14/55 , C07K17/12 , C07K19/00 , C08B15/00 , C08B37/08 , C08H8/00 , C12N9/00 , C12N9/16 , C12N9/38 , C12N9/42 , C12N11/12 , C12N15/09 , C12P21/00 , C12P21/02 , C12Q1/56 , C12R1/19 , D21H17/00 , G01N33/531
CPC分类号： C08H8/00 , A61K39/39 , C07K14/55 , C08B15/00 , C08B37/003 , C12N11/12 , C12N9/16 , C12N9/2437 , C12N9/2445 , C12N9/2468 , C12Q1/56 , C12Y302/01004 , C12Y302/01021 , C12Y302/01091 , D21H17/005 , G01N33/531 , C07K2319/00 , C07K2319/02 , C07K2319/33 , C07K2319/50 , C07K2319/61 , C07K2319/75 , Y10S435/803 , Y10S530/808 , Y10S530/814
摘要： A fusion protein is prepared containing a polypeptide 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 an affinity matrix containing a substrate such as cellulose for the cellulase substrate binding region, the substrate binding region binds to the affinity matrix to immobilize the polypeptide. The polypeptide can be purified by separating the fusion protein or polypeptide from the affinity matrix. The polypeptide can be separated by cleaving the protein with a Cellulomonas fimi protease.
摘要翻译：制备含有多肽的融合蛋白，例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。通过使融合蛋白与含有底物如纤维素的纤维素底物结合区的亲和基质接触，底物结合区结合亲和基质以固定多肽。可以通过将融合蛋白或多肽与亲和基质分离来纯化多肽。多肽可以通过用纤维单胞菌蛋白酶切割蛋白来分离。

6. 发明授权

US06300493B1 Method and compositions for modification of polysaccharide characteristics 有权
标题翻译：用于改性多糖特征的方法和组合物
公开(公告)号：US06300493B1
公开(公告)日：2001-10-09
申请号：US09170545
申请日：1998-10-13
申请人： Neil R. Gilkes , Douglas G. Kilburn , Robert C. Miller, Jr. , Anthony Warren
发明人： Neil R. Gilkes , Douglas G. Kilburn , Robert C. Miller, Jr. , Anthony Warren
IPC分类号： C12P1904
CPC分类号： D06M16/003
摘要： Methods and compositions are provided for the modification of polysaccharide structures using polysaccharidase binding or catalytic domains either alone or in tandem to modify the structure of polysaccharides. These methods and compositions are exemplified by the use of cellulase binding and catalytic domains to polish cotton, and to alter dying characteristics, texture and porosity of cellulose fibers.
摘要翻译：提供了使用多糖酶结合或催化结构域单独或串联修饰多糖结构以修饰多糖结构的方法和组合物。这些方法和组合物的例子是使用纤维素酶结合和催化结构域来抛光棉花，并改变纤维素纤维的染色特性，质地和孔隙率。

7. 发明授权

US5643791A Characterization and structure of an endodglucanase gene of Cellulomonas fimi 失效
标题翻译：葡萄球菌的内切葡聚糖酶基因的表征和结构
公开(公告)号：US5643791A
公开(公告)日：1997-07-01
申请号：US185303
申请日：1994-01-19
申请人： R. Anthony J. Warren , Douglas G. Kilburn , Robert C. Miller, Jr.
发明人： R. Anthony J. Warren , Douglas G. Kilburn , Robert C. Miller, Jr.
IPC分类号： C12N1/21 , C12N9/42 , C12N15/52 , C12N15/63
CPC分类号： C12N9/2437 , C12Y302/01004
摘要： A biologically pure DNA sequence which encodes an endo 1,4-.beta.-glucanase, useful for the efficient conversion of cellulose to glucose, is disclosed. Also disclosed are recombinant DNA cloning vehicles (vectors) that contain nucleotide sequences that encode for the aforesaid endoglucanase, proteins that exhibit endoglucanase activity, expression-controlling DNA sequences, microorganisms that contain recombinant DNA cloning vehicles, as well as messenger RNA sequences complementary to a DNA strand of the aforesaid DNA nucleotide sequences.
摘要翻译：公开了一种生物学纯DNA序列，其编码可用于将纤维素有效转化为葡萄糖的内切1,4-β-葡聚糖酶。还公开了包含编码上述内切葡聚糖酶的核苷酸序列，表现出内切葡聚糖酶活性的蛋白质，表达控制DNA序列，含有重组DNA克隆载体的微生物，以及与其互补的信使RNA序列的重组DNA克隆载体（载体）上述DNA核苷酸序列的DNA链。

8. 发明授权

US5626767A Acoustic filter for separating and recycling suspended particles 失效
标题翻译：用于分离和回收悬浮颗粒的声学过滤器
公开(公告)号：US5626767A
公开(公告)日：1997-05-06
申请号：US88547
申请日：1993-07-02
申请人： Felix Trampler , James M. Piret , Stefan A. Sonderhoff , Douglas G. Kilburn
发明人： Felix Trampler , James M. Piret , Stefan A. Sonderhoff , Douglas G. Kilburn
IPC分类号： B01D21/00 , B01D43/00 , B01J19/10 , G10K11/04 , C02F1/36
CPC分类号： B01D21/283 , B01D43/00 , B01J19/10 , G10K11/04
摘要： Particulate material suspended in a fluid is separated and recycled by means of an ultrasonic resonance wave. In a preferred embodiment, the ultrasonic resonance field is generated within a multilayered composite resonator system including a transducer, the suspension and a mirror parallel to each other. Dimensions and frequencies resonant to the whole system but not exciting Eigen-frequencies of transducer and mirror itself are chosen so that thermal dissipation is minimized. Criteria for flow direction and flow rate are defined in order to maintain a high-quality factor of the composite resonator and to achieve a high-separation efficiency. Generally, the process is suitable for all kinds of particles (solid, liquid or gaseous disperse phases) especially for hydrosols (particles in water) and for separation of biological particles such as mammalian, bacterial and plant cells or aggregates. Specialized applications in biotechnology are described including an acoustic filter for mammalian cell bioreactors or the selective retention of viable cells relative to non-viable cells.
摘要翻译：悬浮在流体中的颗粒物质通过超声共振波被分离和循环。在优选实施例中，在包括换能器，悬架和彼此平行的反射镜的多层复合谐振器系统内产生超声共振场。尺寸和频率与整个系统共振但不激励选择换能器和反射镜本征频率，以使散热最小化。定义了流动方向和流速的标准，以保持复合谐振器的高品质因子并实现高分离效率。通常，该方法适用于各种颗粒（固体，液体或气体分散相），特别是用于水溶胶（水中的颗粒）和分离生物颗粒如哺乳动物，细菌和植物细胞或聚集体。描述了生物技术中的专门应用，包括用于哺乳动物细胞生物反应器的声学过滤器或相对于非活细胞选择性保留活细胞。

9. 发明授权

US5137819A Cellulose binding fusion proteins for immobilization and purification of polypeptides 失效
标题翻译：纤维素结合融合蛋白，用于多肽的固化和纯化
公开(公告)号：US5137819A
公开(公告)日：1992-08-11
申请号：US216794
申请日：1988-07-08
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： C12P19/14 , A61K39/39 , C07K1/22 , C07K14/00 , C07K14/195 , C07K14/41 , C07K14/52 , C07K14/54 , C07K14/55 , C07K17/12 , C07K19/00 , C08B15/00 , C08B37/08 , C08H8/00 , C12N9/00 , C12N9/16 , C12N9/38 , C12N9/42 , C12N11/12 , C12N15/09 , C12P21/00 , C12P21/02 , C12Q1/56 , C12R1/19 , D21H17/00 , G01N33/531
CPC分类号： C08H8/00 , A61K39/39 , C07K14/55 , C08B15/00 , C08B37/003 , C12N11/12 , C12N9/16 , C12N9/2437 , C12N9/2445 , C12N9/2468 , C12Q1/56 , C12Y302/01004 , C12Y302/01021 , C12Y302/01091 , D21H17/005 , G01N33/531 , C07K2319/00 , C07K2319/02 , C07K2319/33 , C07K2319/50 , C07K2319/61 , C07K2319/75 , Y10S435/803 , Y10S530/808 , Y10S530/814
摘要： A fusion protein is prepared containing a polypeptide 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 an affinity matrix containing a substrate such as cellulose for the cellulase substrate binding region, the substrate binding region binds to the affinity matrix to immobilize the polypeptide. The polypeptide can be purified by separating the fusion protein or polypeptide from the affinity matrix.
摘要翻译：制备含有多肽的融合蛋白，例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。通过使融合蛋白与含有底物如纤维素的纤维素底物结合区的亲和基质接触，底物结合区结合亲和基质以固定多肽。可以通过将融合蛋白或多肽与亲和基质分离来纯化多肽。

10. 发明授权

US6124117A Polysaccharide binding fusion proteins and conjugates 失效
标题翻译：多糖结合融合蛋白和缀合物
公开(公告)号：US6124117A
公开(公告)日：2000-09-26
申请号：US788621
申请日：1997-01-23
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 , C12N15/56
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： Novel polypeptide compositions and methods for their use are provided comprising fusion proteins in which the polysaccharide binding domain or functional portion thereof of a polysaccharidase is fused to a heterologous protein or is conjugated to a chemical moiety. The compositions can be synthesized or prepared by recombinant DNA technology. The compositions find use as removable labels.
摘要翻译：提供了新的多肽组合物及其使用方法，其包含融合蛋白，其中多糖酶的多糖结合结构域或其功能部分与异源蛋白融合或与化学部分缀合。可以通过重组DNA技术合成或制备组合物。该组合物用作可移除标签。

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