公开(公告)号：US06906172B2

公开(公告)日：2005-06-14

申请号：US09970150

申请日：2001-10-03

申请人： Barry Bratcher ,  Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch

发明人： Barry Bratcher ,  Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch

IPC分类号： C12M1/00 ,  C07K14/415 ,  C07K16/00 ,  C12M1/10 ,  C12M1/12 ,  C12M1/33 ,  C12M3/00 ,  C12N7/00 ,  C12N7/02 ,  C12N15/82 ,  A61K38/00 ,  C07K1/00

CPC分类号： C12N15/8258 ,  C07K14/415 ,  C07K16/00 ,  C07K2317/13 ,  C07K2319/00 ,  C12N7/00 ,  C12N15/8203 ,  C12N15/8242 ,  C12N15/8257 ,  C12N2710/00051 ,  C12N2750/14151

摘要： A flexible automated apparatus for isolating and purifying viruses, proteins and peptides of interest from a plant material is disclosed, the apparatus being applicable for large scale purification and isolation of such substances from plant material. The flexible automated apparatus provides an efficient apparatus for isolating viruses, proteins and peptides of interest with little waste material. The automated apparatus for isolating viruses, proteins and peptides of interest includes a grinding apparatus for homogenizing a plant to produce a green juice, a means for adjusting the pH of and heating the green juice, a means for separating the target species, either virus or protein/peptide, from other components of the green juice by one or more cycles of centrifugation, resuspension, and ultrafiltration, and finally purifying virus particles by such procedure as PEG-precipitation or purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

摘要翻译： 公开了一种用于从植物材料分离和纯化病毒，蛋白质和肽的灵活的自动化装置，该装置适用于从植物材料大规模纯化和分离这些物质。 灵活的自动化设备提供了一种用于利用少量废料分离病毒，蛋白质和肽的有效装置。 用于分离感兴趣的病毒，蛋白质和肽的自动化装置包括用于使植物均匀化以产生绿汁的研磨装置，用于调节绿汁的pH并加热绿汁的方法，用于分离靶物种的方法，病毒或 蛋白质/肽，通过一个或多个离心循环，再悬浮和超滤，通过诸如PEG沉淀或通过诸如色谱和/或盐的方法纯化蛋白质和肽来纯化病毒颗粒，从绿汁的其它成分 沉淀。

公开(公告)号：US5814495A

公开(公告)日：1998-09-29

申请号：US404384

申请日：1995-03-14

申请人： Guy della-Cioppa ,  Stephen J. Garger, Jr. ,  Genadie G. Sverlow ,  Thomas H. Turpen ,  Laurence K. Grill ,  Miles R. Chedekal

发明人： Guy della-Cioppa ,  Stephen J. Garger, Jr. ,  Genadie G. Sverlow ,  Thomas H. Turpen ,  Laurence K. Grill ,  Miles R. Chedekal

IPC分类号： C07K2/00 ,  C07K14/36 ,  C12N9/02 ,  C12P17/00 ,  C12P17/14 ,  A61K7/44

CPC分类号： C12N9/0059 ,  C07K14/36 ,  C12N9/0071 ,  C12P17/00 ,  C12Y110/03001 ,  C12Y114/18001

摘要： The present invention is directed to a process for producing melanins, their precursors and their analoges, hereinafter referred to generically as melanins. According to the invention, melanins are produced in amounts greater than about 3.3 grams wet weight per liter of growth medium. The enhanced production of melanin can be achieved by manipulating the constituents of the growth medium, and/or attenuating fermentations conditions, and/or by genetically engineering microorganisms to produce melanins, and/or mutating the microorganisms.

摘要翻译： 本发明涉及一种生产黑色素及其前体及其类似物的方法，以下通常称为黑色素。 根据本发明，以每升生长培养基大于约3.3克湿重的量生产黑色素。 可以通过操纵生长培养基的成分和/或减弱发酵条件和/或通过遗传工程微生物产生黑色素和/或使微生物变异来实现黑色素的增强生产。

公开(公告)号：US07034128B2

公开(公告)日：2006-04-25

申请号：US10976698

申请日：2004-10-29

申请人： Thomas H. Turpen ,  Stephen J. Garger ,  Michael J. McCulloch ,  Terri I. Cameron ,  Michelle L. Samonek-Potter ,  R. Barry Holtz

发明人： Thomas H. Turpen ,  Stephen J. Garger ,  Michael J. McCulloch ,  Terri I. Cameron ,  Michelle L. Samonek-Potter ,  R. Barry Holtz

IPC分类号： C07K1/14 ,  C07K1/36

CPC分类号： C12Y302/01045 ,  C07K14/415 ,  C07K14/47 ,  C07K14/57 ,  C07K16/00 ,  C07K2317/13 ,  C07K2317/622 ,  C12N9/2402 ,  C12N9/2422 ,  C12N9/2465 ,  C12N15/8257 ,  C12N15/8258 ,  C12Y302/01022

摘要： A method for extracting proteins from the intercellular space of plants is provided. The method is applicable to the large scale isolation of many active proteins of interest synthesized by plant cells. The method may be used commercially to recover recombinantly produced proteins from plant hosts thereby making the large scale use of plants as sources for recombinant protein production feasible.

公开(公告)号：US06890748B2

公开(公告)日：2005-05-10

申请号：US10103327

申请日：2002-03-20

申请人： Stephen J. Garger ,  Thomas H. Turpen ,  Monto H. Kumagai

发明人： Stephen J. Garger ,  Thomas H. Turpen ,  Monto H. Kumagai

IPC分类号： C07K14/08 ,  C07K14/095 ,  C07K14/415 ,  C07K14/445 ,  C12N9/02 ,  C12N9/10 ,  C12N9/14 ,  C12N9/16 ,  C12N9/18 ,  C12N9/20 ,  C12N9/24 ,  C12N9/40 ,  C12N9/72 ,  C12N9/78 ,  C12N9/84 ,  C12N15/40 ,  C12N15/82 ,  C12N15/83 ,  C12N15/86 ,  C12P41/00 ,  C07H21/04 ,  C07K17/00 ,  C12P21/06 ,  C12Q1/34

CPC分类号： C12Y302/01045 ,  C07K14/005 ,  C07K14/415 ,  C07K14/445 ,  C07K2319/00 ,  C12N9/0059 ,  C12N9/0071 ,  C12N9/1074 ,  C12N9/14 ,  C12N9/16 ,  C12N9/18 ,  C12N9/20 ,  C12N9/2402 ,  C12N9/2465 ,  C12N9/6459 ,  C12N9/78 ,  C12N9/84 ,  C12N15/8203 ,  C12N15/8216 ,  C12N15/8242 ,  C12N15/8257 ,  C12N15/8289 ,  C12N15/86 ,  C12N2770/00022 ,  C12N2770/32722 ,  C12P41/003 ,  C12Y114/18001 ,  C12Y302/01022 ,  C12Y302/01031 ,  C12Y304/21069

摘要： The invention relates to α-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human α galactosidase nucleotide sequences. The recombinant lysosomal enzymes produced in accordance with the invention may be used for a variety of purposes, including but not limited to enzyme replacement therapy for the therapeutic treatment of human and animal lysosomal storage diseases.

摘要翻译： 本发明涉及在羧基末端截短的α-半乳糖苷酶和生产酶活性重组人和动物溶酶体酶，其涉及重组表达构建体的构建和表达，其包含在植物表达系统中编码人或动物溶酶体酶的序列。 植物表达系统提供翻译后修饰和加工以产生显示酶活性的重组基因产物。 本发明通过其中转基因烟草植物表达包含人葡糖脑苷脂酶核苷酸序列的重组表达构建体的实施例来证明。 本发明还通过其中转染的烟草植物表达包含人α半乳糖苷酶核苷酸序列的重组病毒表达构建体的实施例来证明。 根据本发明生产的重组溶酶体酶可用于多种目的，包括但不限于用于治疗性治疗人和动物溶酶体贮积病的酶替代疗法。

公开(公告)号：US06887696B2

公开(公告)日：2005-05-03

申请号：US09993059

申请日：2001-11-13

申请人： Stephen J. Garger ,  Thomas H. Turpen ,  Monto H. Kumagai

发明人： Stephen J. Garger ,  Thomas H. Turpen ,  Monto H. Kumagai

IPC分类号： C07K14/08 ,  C07K14/095 ,  C07K14/415 ,  C07K14/445 ,  C12N9/02 ,  C12N9/10 ,  C12N9/14 ,  C12N9/16 ,  C12N9/18 ,  C12N9/20 ,  C12N9/24 ,  C12N9/40 ,  C12N9/72 ,  C12N9/78 ,  C12N9/84 ,  C12N15/40 ,  C12N15/82 ,  C12N15/83 ,  C12N15/86 ,  C12P41/00 ,  C07K17/00 ,  C12P21/06 ,  C12Q1/34

CPC分类号： C12Y302/01045 ,  C07K14/005 ,  C07K14/415 ,  C07K14/445 ,  C07K2319/00 ,  C12N9/0059 ,  C12N9/0071 ,  C12N9/1074 ,  C12N9/14 ,  C12N9/16 ,  C12N9/18 ,  C12N9/20 ,  C12N9/2402 ,  C12N9/2465 ,  C12N9/6459 ,  C12N9/78 ,  C12N9/84 ,  C12N15/8203 ,  C12N15/8216 ,  C12N15/8242 ,  C12N15/8257 ,  C12N15/8289 ,  C12N15/86 ,  C12N2770/00022 ,  C12N2770/32722 ,  C12P41/003 ,  C12Y114/18001 ,  C12Y302/01022 ,  C12Y302/01031 ,  C12Y304/21069

摘要： The invention relates to α-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human α galactosidase nucleotide sequences. The recombinant lysosomal enzymes produced in accordance with the invention may be used for a variety of purposes, including but not limited to enzyme replacement therapy for the therapeutic treatment of human and animal lysosomal storage diseases.

公开(公告)号：US06037456A

公开(公告)日：2000-03-14

申请号：US37751

申请日：1998-03-10

申请人： Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

发明人： Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

IPC分类号： C12N15/09 ,  C07K14/415 ,  C07K16/00 ,  C07K16/16 ,  C12N7/00 ,  C12N7/02 ,  C12N15/82 ,  C07K1/14 ,  A61K35/78 ,  C07H1/08 ,  C07K1/36

CPC分类号： C12N15/8258 ,  C07K14/415 ,  C07K16/00 ,  C07K16/16 ,  C12N15/8203 ,  C12N15/8242 ,  C12N15/8257 ,  C12N7/00 ,  C07K2317/13 ,  C07K2319/00 ,  C12N2710/00051 ,  C12N2770/00051 ,  Y10S977/904

摘要： The present invention features a method for isolating and purifying viruses, proteins and peptides of interest from a plant host which is applicable on a large scale. Moreover, the present invention provides a more efficient method for isolating viruses, proteins and peptides of interest than those methods described in the prior art. In general, the present method of isolating viruses, proteins and peptides of interest comprises the steps of homogenizing a plant to produce a green juice, adjusting the pH of and heating the green juice, separating the target species, either virus or protein/peptide, from other components of the green juice by one or more cycles of centrifugation, resuspenion, and ultrafiltration, and finally purifying virus particles by such procedure as PEG-precipitation or purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

摘要翻译： 本发明的特征在于从植物宿主分离和纯化病毒，蛋白质和肽的方法，该方法可大规模应用。 此外，本发明提供了一种比现有技术中描述的方法更有效的分离病毒，蛋白质和肽的方法。 通常，目前分离感兴趣的病毒，蛋白质和肽的方法包括以下步骤：将植物均质化以产生绿汁，调节绿汁的pH并加热绿汁，分离靶物种，病毒或蛋白质/肽， 通过一次或多次离心，再次和超滤循环，从绿汁的其它组分中，最后通过诸如PEG沉淀或通过诸如色谱法和/或盐沉淀的方法纯化蛋白质和肽来纯化病毒颗粒。

公开(公告)号：US5340734A

公开(公告)日：1994-08-23

申请号：US982095

申请日：1992-11-25

申请人： Guy R. della-Cioppa ,  Stephen J. Garger, Jr. ,  Richard B. Holtz ,  Michael J. McCulloch ,  Genadie G. Sverlow

发明人： Guy R. della-Cioppa ,  Stephen J. Garger, Jr. ,  Richard B. Holtz ,  Michael J. McCulloch ,  Genadie G. Sverlow

IPC分类号： C12N15/09 ,  C08G61/00 ,  C08H99/00 ,  C12N20060101 ,  C12N1/21 ,  C12N9/02 ,  C12N9/96 ,  C12N15/53 ,  C12P20060101 ,  C12P7/00 ,  C12P17/00 ,  C12P21/08 ,  C12R1/48 ,  C12N9/48 ,  C08G59/68 ,  C12P17/06

CPC分类号： C12Y110/03001 ,  C08G61/00 ,  C12N9/0059 ,  C12N9/0071 ,  C12P17/00 ,  C12Y114/18001

摘要： A process for the in vitro production of chemically modified polyphenolic polymer (PPP). First, stable, highly active extracellular tyrosinase is produced from genetically transformed microorganism such as Streptomyces antibioticus. The tyrosinase is then incubated with a reaction substrate such as l-tyrosine, hydrolyzed protein, or an oligopeptide in combination with l-tyrosine. The ratio of the oligopeptide/tyrosine combination as well as variation in the concentration of tyrosinase can be used to modify the color, the molecular size, and the spectral absorbance properties of the PPP produced. Alternatively, or additionally, oxidants such as hydrogen peroxide or hypochlorite can be used to modify the color of the PPP, regardless of the method used to produce the PPP, and the PPP can subsequently be fractionated using molecular weight cut-off ultrafiltration. Organic solvents can also be used in the method of making PPP to produce PPPs having variable but reproducible physical properties.

摘要翻译： 一种体外生产化学改性多酚聚合物（PPP）的方法。 首先，稳定的，高活性的细胞外酪氨酸酶由遗传转化的微生物如抗链霉菌（Streptomyces antibioticus）产生。 然后将酪氨酸酶与反应底物如1-酪氨酸，水解蛋白质或寡肽与1-酪氨酸组合孵育。 寡肽/酪氨酸组合的比例以及酪氨酸酶浓度的变化可用于改变生产的PPP的颜色，分子大小和光谱吸收性质。 或者或另外，可以使用氧化剂如过氧化氢或次氯酸盐来改变PPP的颜色，而不管用于生产PPP的方法如何，并且随后可以使用分子量截止超滤对PPP进行分级。 有机溶剂也可用于制备PPP以产生具有可变但可再现的物理性质的PPP的方法。

公开(公告)号：US07192740B2

公开(公告)日：2007-03-20

申请号：US10280725

申请日：2002-10-24

申请人： Jonathan Donson ,  William O. Dawson ,  George L. Grantham ,  Thomas H. Turpen ,  Ann Myers Turpen ,  Stephen J. Garger, Jr. ,  Laurence K. Grill

发明人： Jonathan Donson ,  William O. Dawson ,  George L. Grantham ,  Thomas H. Turpen ,  Ann Myers Turpen ,  Stephen J. Garger, Jr. ,  Laurence K. Grill

IPC分类号： C12P21/02 ,  C12P21/06 ,  C12N15/00 ,  C12N15/74

CPC分类号： C07K14/005 ,  C07K14/415 ,  C12N9/00 ,  C12N9/0059 ,  C12N9/0071 ,  C12N9/0083 ,  C12N9/1074 ,  C12N9/14 ,  C12N9/16 ,  C12N9/18 ,  C12N9/20 ,  C12N9/6459 ,  C12N9/78 ,  C12N9/84 ,  C12N15/1137 ,  C12N15/8203 ,  C12N15/8216 ,  C12N15/8218 ,  C12N15/8249 ,  C12N15/825 ,  C12N15/8289 ,  C12N15/86 ,  C12N2710/22043 ,  C12N2710/22062 ,  C12N2750/12043 ,  C12N2770/00022 ,  C12N2770/00043 ,  C12N2770/00051 ,  C12N2770/32643 ,  C12N2770/32662 ,  C12N2770/32722 ,  C12N2770/32743 ,  C12N2770/32762 ,  C12N2770/36122 ,  C12N2770/36143 ,  C12N2770/36162 ,  C12N2810/609 ,  C12N2830/00 ,  C12N2830/60 ,  C12P41/003 ,  C12Y114/18001 ,  C12Y302/01031 ,  C12Y304/21069

摘要： The present invention relates to a recombinant viral nucleic acid selected from a (+) sense, single stranded RNA virus possessing a native subgenomic promoter encoding for a first viral subgenomic promoter, a nucleic acid sequence that codes for a viral coat protein whose transcription is regulated by the first viral subgenomic promoter, a second viral subgenomic promoter and a second nucleic acid sequence whose transcription is regulated by the second viral subgenomic promoter. The first and second viral subgenomic promoters of the recombinant viral nucleic acid do not have homologous sequences relative to each other. The recombinant viral nucleic acid provides the particular advantage that it systemically transcribes the second nucleic acid in the host. Host organisms encompassed by the present invention include procaryotes and eucaryotes, particularly animals and plants. The present invention also relates to viruses containing the viral vectors which are infective, production cells which are capable of producing the viruses or parts thereof, a host infected by the viruses of the invention, the gene products produced by expression of the viral nucleic acids and a process for the production of a desired product by growing the infected hosts.

摘要翻译： 本发明涉及选自具有编码第一病毒亚基因组启动子的天然亚基因启动子的（+）有义单链RNA病毒的重组病毒核酸，编码转录调节的病毒外壳蛋白的核酸序列 通过第一病毒亚基因组启动子，第二病毒亚基因组启动子和其转录受第二病毒亚基因组启动子调控的第二核酸序列。 重组病毒核酸的第一和第二病毒亚基因组启动子相对于彼此不具有同源序列。 重组病毒核酸提供其系统地转录宿主中的第二核酸的特别优点。 本发明包括的宿主生物包括原核生物和真核生物，特别是动物和植物。 本发明还涉及含有感染性病毒载体的病毒，能够产生病毒或其部分的生产细胞，被本发明的病毒感染的宿主，通过表达病毒核酸产生的基因产物和 通过生长受感染的宿主来生产所需产品的方法。

公开(公告)号：US06841659B2

公开(公告)日：2005-01-11

申请号：US10632240

申请日：2003-08-01

申请人： Thomas H. Turpen ,  Stephen J. Garger ,  Michael J. McCulloch ,  Terri I. Cameron ,  Michelle L. Samonek-Potter ,  R. Barry Holtz

发明人： Thomas H. Turpen ,  Stephen J. Garger ,  Michael J. McCulloch ,  Terri I. Cameron ,  Michelle L. Samonek-Potter ,  R. Barry Holtz

IPC分类号： C12N15/09 ,  A61K36/81 ,  B01D15/08 ,  B01D61/14 ,  B01J3/00 ,  C07K1/16 ,  C07K1/34 ,  C07K14/415 ,  C07K14/47 ,  C07K14/57 ,  C07K16/00 ,  C07K16/16 ,  C12N9/14 ,  C12N9/24 ,  C12N9/32 ,  C12N9/40 ,  C12N15/82 ,  C07H1/06 ,  C07H1/08 ,  C07K1/14 ,  C11B1/04

CPC分类号： C12Y302/01045 ,  C07K14/415 ,  C07K14/47 ,  C07K14/57 ,  C07K16/00 ,  C07K2317/13 ,  C07K2317/622 ,  C12N9/2402 ,  C12N9/2422 ,  C12N9/2465 ,  C12N15/8257 ,  C12N15/8258 ,  C12Y302/01022

摘要： A method for extracting proteins from the intercellular space of plants is provided. The method is applicable to the large scale isolation of many active proteins of interest synthesized by plant cells. The method may be used commercially to recover recombinantly produced proteins from plant hosts thereby making the large scale use of plants as sources for recombinant protein production feasible.

公开(公告)号：US06740740B2

公开(公告)日：2004-05-25

申请号：US09962527

申请日：2001-09-24

申请人： Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

发明人： Stephen J. Garger ,  R. Barry Holtz ,  Michael J. McCulloch ,  Thomas H. Turpen

IPC分类号： C07K136

CPC分类号： C12N15/8258 ,  C07K14/415 ,  C07K16/00 ,  C07K16/16 ,  C07K2317/13 ,  C07K2319/00 ,  C12N7/00 ,  C12N15/8203 ,  C12N15/8242 ,  C12N15/8257 ,  C12N2710/00051 ,  C12N2770/00051 ,  Y10S977/904

摘要： The present invention features a method for isolating and purifying proteins and peptides of interest from a plant host, which is applicable on a larger scale. Moreover, the present invention provides a more efficient method for isolating proteins and peptides of interest than those methods described in the prior art. In general, the present method of isolating proteins and peptides of interest comprises the steps of homogenizing a plant to produce a green juice, adjusting the pH of and heating the green juice, separating the target protein/peptide from other components of the green juice by one or more cycles of centrifugation and/or resuspension, and finally purifying proteins and peptides by such procedures as chromatography and/or salt precipitation.

摘要翻译： 本发明特征在于从植物宿主中分离和纯化目的蛋白质和肽的方法，该方法可适用于较大规模。 此外，本发明提供了一种比现有技术中描述的方法更有效的分离蛋白质和肽的方法。 通常，本发明的分离蛋白质和肽的方法包括以下步骤：将植物均质化以产生绿汁，调节绿汁的pH并加热绿汁，将目标蛋白质/肽与绿汁的其它成分分离，通过 离心和/或再悬浮的一个或多个循环，并且最后通过诸如色谱法和/或盐沉淀的方法纯化蛋白质和肽。