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    COMPOSITIONS AND METHODS FOR HUMANIZATION AND OPTIMIZATION OF N-GLYCANS IN PLANTS
    2.
    发明申请
    COMPOSITIONS AND METHODS FOR HUMANIZATION AND OPTIMIZATION OF N-GLYCANS IN PLANTS 审中-公开
    N-GLYCANS在植物中的人类化和优化的组合物和方法

    公开(公告)号：US20110144308A1

    公开(公告)日：2011-06-16

    申请号：US12955611

    申请日：2010-11-29

    申请人： LYNN F. DICKEY ,  KEVIN M. COX ,  CHARLES G. PEELE

    发明人： LYNN F. DICKEY ,  KEVIN M. COX ,  CHARLES G. PEELE

    IPC分类号： C07K16/00

    CPC分类号： C07K14/47 ,  C07K16/00 ,  C07K2317/13 ,  C07K2317/41 ,  C07K2317/71 ,  C07K2317/72 ,  C07K2317/732 ,  C07K2317/734 ,  C12N15/8257 ,  C12N15/8258

    摘要： Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant construct that provides for the inhibition of expression of α1,3-fucosyltransferase (FucT) and β1,2-xylosyltransferase (XylT) in a plant. Use of these constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

    摘要翻译： 提供了改变高等植物中蛋白质N-糖基化模式的方法。 所述方法包括向植物中引入一种重组构建体，其在植物中提供抑制α1,3-岩藻糖基转移酶（FucT）和（b），1,2-木糖基转移酶（XylT）的表达。 这些构建体用于抑制或抑制这两种酶及其同种型的表达有利地提供具有“人源化”N-糖基化模式的内源和异源蛋白质的生产，而不影响植物的生长和发育。 提供了具有这种蛋白质N-糖基化模式的稳定转化的高等植物。 还提供了具有基本上均匀的糖基化曲线并且对于G0糖型基本均匀的糖蛋白组合物，包括单克隆抗体组合物。

    Expression of plasminogen and microplasminogen in duck weed
    3.
    发明授权
    Expression of plasminogen and microplasminogen in duck weed 失效
    在鸭杂草中表达纤溶酶原和微量淀粉样蛋白

    公开(公告)号：US07659445B2

    公开(公告)日：2010-02-09

    申请号：US11056621

    申请日：2005-02-11

    申请人： David Spencer ,  Lynn F. Dickey ,  John R. Gasdaska ,  Xiaowei Wang ,  Kevin M. Cox ,  Charles G. Peele

    发明人： David Spencer ,  Lynn F. Dickey ,  John R. Gasdaska ,  Xiaowei Wang ,  Kevin M. Cox ,  Charles G. Peele

    IPC分类号： C12N15/82 ,  C12N15/12 ,  C12N15/62

    CPC分类号： C12N15/8257 ,  C12N9/6435 ,  C12Y304/21007

    摘要： The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

    摘要翻译： 本发明提供了用于在浮萍表达系统中生产重组纤溶酶原，微量血浆​​原和其片段的方法和组合物。 本发明的新发现是，浮萍表达系统可用于产生高水平的纤溶酶原和微量淀粉样蛋白。 可以将浮萍产生的纤溶酶原和微量血浆原活化以产生具有蛋白酶活性的多肽。 因此，本发明包括在用于表达纤溶酶原，微量血浆​​原和其片段的表达盒转化的浮萍，浮萍植物中表达纤溶酶原，微量淀粉样蛋白及其片段的方法，以及包含编码纤溶酶原，微量稀释细胞的核苷酸序列的核酸 及其片段，其中这些核苷酸序列被修饰以增强其在浮萍中的表达。

    GLYCAN-OPTIMIZED ANTI-CD20 ANTIBODIES
    4.
    发明申请
    GLYCAN-OPTIMIZED ANTI-CD20 ANTIBODIES 审中-公开
    GLYCAN优化的抗CD20抗体

    公开(公告)号：US20090060921A1

    公开(公告)日：2009-03-05

    申请号：US12115133

    申请日：2008-05-05

    申请人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele ,  Ming-Bo Wang

    发明人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele ,  Ming-Bo Wang

    IPC分类号： A61K39/395 ,  C07K16/18 ,  C12N5/04 ,  A61P31/00

    CPC分类号： C07K14/47 ,  C07K16/00 ,  C07K16/2878 ,  C07K16/2887 ,  C07K2317/13 ,  C07K2317/41 ,  C07K2317/71 ,  C07K2317/72 ,  C07K2317/732 ,  C07K2317/734 ,  C12N15/8257 ,  C12N15/8258

    摘要： Glycan-optimized monoclonal antibodies that specifically bind CD20 antigen and which have improved effector function are provided. The anti-CD20 antibodies of the invention have a glycosylation pattern that results in an antibody composition having predominately the G0 glycoform, and thus comprise N-glycans that lack fucose (i.e., afucosylated) and galactose residues attached thereto. In some embodiments, these anti-CD20 antibodies comprise the light chain and heavy chain sequences of the rituximab anti-CD20 antibody, and thus represent afucosylated rituximab. Methods for producing these glycan-optimized anti-CD20 antibodies are also provided.

    摘要翻译： 提供特异性结合CD20抗原并具有改善的效应子功能的糖基化优化的单克隆抗体。 本发明的抗CD20抗体具有糖基化模式，其导致抗体组合物主要具有G0糖型，并且因此包含缺少岩藻糖（即非岩藻糖基化）的N-聚糖和与其连接的半乳​​糖残基。 在一些实施方案中，这些抗CD20抗体包含利妥昔单抗抗CD20抗体的轻链和重链序列，因此代表非岩藻糖基化的利妥昔单抗。 还提供了生产这些聚糖优化的抗CD20抗体的方法。

    Compositions and methods for inhibition of fucosyltransferase and xylosyltransferase expression in plants
    5.
    发明授权
    Compositions and methods for inhibition of fucosyltransferase and xylosyltransferase expression in plants 失效
    用于抑制植物中岩藻糖基转移酶和木糖基转移酶表达的组合物和方法

    公开(公告)号：US08716557B2

    公开(公告)日：2014-05-06

    申请号：US11624158

    申请日：2007-01-17

    申请人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele ,  Ming-Bo Wang

    发明人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele ,  Ming-Bo Wang

    IPC分类号： C12N15/82 ,  C07H21/04 ,  A01H5/00

    CPC分类号： C07K14/47 ,  C07K16/00 ,  C07K2317/13 ,  C07K2317/41 ,  C07K2317/71 ,  C07K2317/72 ,  C07K2317/732 ,  C07K2317/734 ,  C12N15/8257 ,  C12N15/8258

    摘要： Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant RNAi construct that provides for the inhibition of expression of α1,3-fucosyltransferase (FucT) and β1,2-xylosyltransferase (XylT) in a plant. Use of these RNAi constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

    摘要翻译： 提供了改变高等植物中蛋白质N-糖基化模式的方法。 所述方法包括将植物中的α1,3-岩藻糖基转移酶（FucT）和β-木糖基转移酶（XylT）的表达抑制的重组RNAi构建体引入植物中。 这些RNAi构建体用于抑制或抑制这两种酶及其同种型的表达有利地提供具有“人源化”N-糖基化模式的内源和异源蛋白质的生产而不影响植物的生长和发育。 提供了具有这种蛋白质N-糖基化模式的稳定转化的高等植物。 还提供了具有基本上均匀的糖基化曲线并且对于G0糖型基本均匀的糖蛋白组合物，包括单克隆抗体组合物。

    Expression of plasminogen and microplasminogen in duckweed
    6.
    发明授权
    Expression of plasminogen and microplasminogen in duckweed 失效
    浮萍中纤维蛋白溶酶原和微量淀粉样蛋白的表达

    公开(公告)号：US08017836B2

    公开(公告)日：2011-09-13

    申请号：US12644640

    申请日：2009-12-22

    申请人： David Spencer ,  Lynn F. Dickey ,  John R. Gasdaska ,  Xiaowei Wang ,  Kevin M. Cox ,  Charles G. Peele

    发明人： David Spencer ,  Lynn F. Dickey ,  John R. Gasdaska ,  Xiaowei Wang ,  Kevin M. Cox ,  Charles G. Peele

    IPC分类号： A01H5/00 ,  C12N15/82 ,  C12N5/04 ,  C12N5/10 ,  C12N15/12 ,  C12N15/62

    CPC分类号： C12N15/8257 ,  C12N9/6435 ,  C12Y304/21007

    摘要： The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

    摘要翻译： 本发明提供了用于在浮萍表达系统中生产重组纤溶酶原，微量血浆​​原和其片段的方法和组合物。 本发明的新发现是，浮萍表达系统可用于产生高水平的纤溶酶原和微量淀粉样蛋白。 可以将浮萍产生的纤溶酶原和微量血浆原活化以产生具有蛋白酶活性的多肽。 因此，本发明包括在用于表达纤溶酶原，微量血浆​​原和其片段的表达盒转化的浮萍，浮萍植物中表达纤溶酶原，微量淀粉样蛋白及其片段的方法，以及包含编码纤溶酶原，微量稀释细胞的核苷酸序列的核酸 及其片段，其中这些核苷酸序列被修饰以增强其在浮萍中的表达。

    EXPRESSION OF PLASMINOGEN AND MICROPLASMINOGEN IN DUCKWEED
    7.
    发明申请
    EXPRESSION OF PLASMINOGEN AND MICROPLASMINOGEN IN DUCKWEED 失效
    表达胰岛素和微管蛋白在DUCKWEED中的表达

    公开(公告)号：US20100186126A1

    公开(公告)日：2010-07-22

    申请号：US12644640

    申请日：2009-12-22

    申请人： DAVID SPENCER ,  LYNN F. DICKEY ,  JOHN R. GASDASKA ,  XIAOWEI WANG ,  KEVIN M. COX ,  CHARLES G. PEELE

    发明人： DAVID SPENCER ,  LYNN F. DICKEY ,  JOHN R. GASDASKA ,  XIAOWEI WANG ,  KEVIN M. COX ,  CHARLES G. PEELE

    IPC分类号： A01H5/00 ,  C12N5/10 ,  C12N9/68

    CPC分类号： C12N15/8257 ,  C12N9/6435 ,  C12Y304/21007

    摘要： The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

    摘要翻译： 本发明提供了用于在浮萍表达系统中生产重组纤溶酶原，微量血浆​​原和其片段的方法和组合物。 本发明的新发现是，浮萍表达系统可以用于产生高水平的纤溶酶原和微量淀粉样蛋白。 可以将浮萍产生的纤溶酶原和微量血浆原活化以产生具有蛋白酶活性的多肽。 因此，本发明包括在用于表达纤溶酶原，微量血浆​​原和其片段的表达盒转化的浮萍，浮萍植物中表达纤溶酶原，微量淀粉样蛋白及其片段的方法，以及包含编码纤溶酶原，微量稀释细胞的核苷酸序列的核酸 及其片段，其中这些核苷酸序列被修饰以增强其在浮萍中的表达。

    COMPOSITIONS AND METHODS FOR PRODUCTION OF AGLYCOSYLATED PLASMINOGEN
    9.
    发明申请
    COMPOSITIONS AND METHODS FOR PRODUCTION OF AGLYCOSYLATED PLASMINOGEN 审中-公开
    用于生产合成的PLASMINOGEN的组合物和方法

    公开(公告)号：US20120220015A1

    公开(公告)日：2012-08-30

    申请号：US13321719

    申请日：2010-05-26

    申请人： Lynn F. Dickey ,  John R. Gasdaska ,  Kevin M. Cox

    发明人： Lynn F. Dickey ,  John R. Gasdaska ,  Kevin M. Cox

    IPC分类号： C12N9/68 ,  C12N5/10 ,  A01H5/00 ,  C12N15/57 ,  C12N15/82

    CPC分类号： A61K38/168 ,  C12N9/6435 ,  C12N15/8257 ,  C12Y304/21007

    摘要： Compositions and methods for producing aglycosylated plasminogen (PLG) polypeptides and fragments and variants thereof are provided. Compositions of the invention include isolated nucleic acid molecules encoding aglycosylated PLG polypeptides in which the asparagine (Asn) residue corresponding to residue Asn-289 of the mature human PLG polypeptide has been substituted with an amino acid residue that does not support N-linked glycosylation at that position of the PLG polypeptide, as well as the aglycosylated PLG polypeptides encoded thereby. Expression constructs comprising these PLG-encoding nucleic acid molecules and transgenic plants comprising these expression constructs are also provided. Methods of the invention comprise introducing a PLG-encoding nucleic acid molecule of the invention into a plant of interest and culturing the plant under conditions to produce the aglycosylated PLG polypeptide. The aglycosylated PLG polypeptide allows for significant increases in production and yield of PLG from a plant-based expression system without comprising the ability of the PLG product to be activated to a polypeptide capable of binding fibrin and having serine protease activity, including biologically active plasmin that is also glycosylated. The activated aglycosylated plasmin is useful to treat diseases or conditions associated with a thrombus.

    摘要翻译： 提供了用于产生糖基化纤溶酶原（PLG）多肽及其片段和变体的组合物和方法。 本发明的组合物包括分离的编码糖基化PLG多肽的核酸分子，其中对应于成熟人PLG多肽的残基Asn-289的天冬酰胺（Asn）残基已被不支持N-连接糖基化的氨基酸残基取代 PLG多肽的位置，以及由此编码的糖基化的PLG多肽。 还提供了包含编码这些编码PLG的核酸分子和包含这些表达构建体的转基因植物的表达构建体。 本发明的方法包括将本发明的编码PLG的核酸分子引入感兴趣的植物中，并在产生糖苷基化的PLG多肽的条件下培养该植物。 糖基化的PLG多肽允许来自基于植物的表达系统的PLG的产量和产量的显着增加，而不包括PLG产物被活化到能够结合纤维蛋白并具有丝氨酸蛋白酶活性的多肽的能力，包括生物活性纤溶酶， 也被糖基化。 活化的糖基化纤溶酶可用于治疗与血栓相关的疾病或病症。

    Expression control elements from the Lemnaceae family
    10.
    发明授权
    Expression control elements from the Lemnaceae family 失效
    来自Lemnaceae科的表达控制元件

    公开(公告)号：US08034916B2

    公开(公告)日：2011-10-11

    申请号：US12581752

    申请日：2009-10-19

    申请人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele

    发明人： Lynn F. Dickey ,  Kevin M. Cox ,  Charles G. Peele

    IPC分类号： C07H21/04 ,  A01H5/00 ,  C12N15/05 ,  C12N15/82

    CPC分类号： C12N15/8216 ,  C07K14/415 ,  C12N9/2442 ,  C12N9/88 ,  C12N15/8221 ,  C12N15/8222 ,  C12N15/8223 ,  C12Y302/01014 ,  C12Y401/01039

    摘要： Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby. Where an expression construct of the invention is designed to express a polypeptide of interest, this novel signal peptide-encoding sequence can be included within the expression construct of the invention to provide for extracellular secretion of the encoded polypeptide of interest.

    摘要翻译： 提供了用于调节植物中感兴趣的核苷酸序列的表达的组合物和方法。 组合物包括新的核酸分子及其变体和片段，用于从莲子科泛素，r-组蛋白和几丁质酶基因分离的表达控制元件。 还提供了使用本文公开的表达控制元件在植物中表达目标核苷酸序列的方法。 该方法包括向植物或植物细胞或结节引入包含与感兴趣的核苷酸序列可操作地连接的本发明的表达控制元件的表达构建体。 特别地，组合物和方法可用于增强浮萍中感兴趣的核苷酸序列的表达。 还提供了一种新颖的莲子科信号肽编码序列和由其编码的信号肽。 当本发明的表达构建体设计用于表达感兴趣的多肽时，该新型信号肽编码序列可以包括在本发明的表达构建体内，以提供所编码的目标多肽的细胞外分泌。