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    GENERATION OF ANTIGENIC VIRUS-LIKE PARTICLES THROUGH PROTEIN-PROTEIN LINKAGES
    1.
    发明申请
    GENERATION OF ANTIGENIC VIRUS-LIKE PARTICLES THROUGH PROTEIN-PROTEIN LINKAGES 审中-公开
    通过蛋白质蛋白链接产生抗原性病毒颗粒

    公开(公告)号：WO2011057134A1

    公开(公告)日：2011-05-12

    申请号：PCT/US2010/055717

    申请日：2010-11-05

    Applicant: NOVICI BIOTECH LLC ,  PADGETT, Hal

    Inventor： PADGETT, Hal

    IPC: C12P21/06 ,  A61K9/14

    CPC classification number: A61K39/21 ,  C07K14/005 ,  C07K14/43581 ,  C07K14/70596 ,  C07K2319/00 ,  C12N7/00 ,  C12N2770/00022 ,  C12N2770/00023

    Abstract: We have generated virus-like particles (VLPs) that can display other proteins through covalent protein-protein linkages mediated by the 'Dock and Lock' interaction between the Drosophila NorpA protein and the C-terminal pentapeptide tail of the InaD protein. This interaction may also be mediated by a portion of the SITAC protein and the Tetraspanin L6 Antigen protein. This system can be used to generate high-density scaffolded arrays of epitopes for immunization. This technology can streamline VLP vaccine candidate production, making it possible to rapidly evaluate panels of candidates in response to current vaccine needs and emerging pathogen threats.

    Abstract translation: 我们已经产生病毒样颗粒（VLPs），可以通过果蝇NorpA蛋白和InaD蛋白的C末端五肽尾部之间的“Dock和Lock”相互作用介导的共价蛋白质 - 蛋白质键显示其他蛋白质。 这种相互作用也可能由SITAC蛋白和四肽蛋白L6抗原蛋白的一部分介导。 该系统可用于产生用于免疫的表位的高密度支架阵列。 该技术可以简化VLP疫苗候选生产，可以根据目前的疫苗需求和新出现的病原体威胁快速评估候选人小组。

    MODIFIED TOBACCO MOSAIC VIRUS PARTICLES AS SCAFFOLDS FOR DISPLAY OF PROTEIN ANTIGENS FOR VACCINE APPLICATIONS
    3.
    发明申请
    MODIFIED TOBACCO MOSAIC VIRUS PARTICLES AS SCAFFOLDS FOR DISPLAY OF PROTEIN ANTIGENS FOR VACCINE APPLICATIONS 审中-公开
    作为疫苗显示用于疫苗应用的蛋白质抗原的改良烟草马赛克病毒颗粒

    公开(公告)号：WO2007038145A2

    公开(公告)日：2007-04-05

    申请号：PCT/US2006/036668

    申请日：2006-09-08

    Applicant: LARGE SCALE BIOLOGY CORPORATION

    Inventor： LINDBO, John, A. ,  SMITH, Mark, L. ,  PALMER, Kenneth, E.

    IPC: C12N7/00

    CPC classification number: C07K14/005 ,  A61K39/12 ,  A61K39/21 ,  A61K39/385 ,  A61K2039/5258 ,  A61K2039/55566 ,  A61K2039/6075 ,  A61K2039/625 ,  C07K2319/00 ,  C07K2319/22 ,  C07K2319/60 ,  C12N7/00 ,  C12N15/8203 ,  C12N15/8257 ,  C12N15/8258 ,  C12N2710/20022 ,  C12N2710/20023 ,  C12N2710/20034 ,  C12N2740/16122 ,  C12N2740/16134 ,  C12N2770/00022

    Abstract: Display of peptides or proteins in an ordered, repetitive array, such as on the surface of a virus- like particle, is known to induce an enhanced immune response relative to vaccination with the "free" protein antigen. The 2100 coat proteins comprising the rod-shaped capsid of Tobacco mosaic virus (TMV) can accommodate short peptide insertions into the primary sequence, but the display of larger protein moieties on the virion surface by genetic fusions to the capsid protein has not been possible. Since TMV lacks surface exposed residues compatible with commonly available linker chemistries, we employed a randomized library approach to introduce a reactive lysine at the externally located at the amino-terminus of the coat protein. We found that we could easily control the extent of virion conjugation and demonstrated stoichiometric biotinylation of the introduced lysine. To characterize this modular platform for the display of heterologous proteins, we bound a model antigen (streptavidin (S A)-green fluorescent protein (GFP), expressed and purified from plants) to the surface of TMV, creating a GFP-SA decorated virus particle. Rapid and quantitative determination of the level of TMV capsid decoration was accomplished by subjecting the complex to amino acid analysis and solving the family of linear equations relating the pmoles of each residue to the known amino acid composition of the complex components. We obtained a GFP-SA tetramer loading of 26%, which corresponds to display of approximately 2200 GFP moieties per intact virion. We evaluated the immunogenicity of GFP decorated virions in both mice and guinea pigs, and found augmented humoral IgG titers in both species, relative to unbound GFP-SA tetramer. In mice, we observed a detectable humoral immune response after only a single immunization with the TMV-protein complex. By demonstrating the presentation of whole proteins, this study expands the utility of TMV as a vaccine scaffold beyond that which is possible by genetic manipulation.

    Abstract translation: 已知有序重复阵列（例如病毒样颗粒表面）上显示肽或蛋白质相对于使用“游离”蛋白质抗原的疫苗接种诱导增强的免疫应答。 包含烟草花叶病毒（TMV）的棒状衣壳的2100涂层蛋白可以适应短的肽插入到一级序列中，但通过与衣壳蛋白的遗传融合在病毒粒子表面上显示更大的蛋白质部分是不可能的。 由于TMV缺乏与通常可用的接头化学物质相容的表面暴露残留物，因此我们采用随机文库方法在外部位于外壳蛋白的氨基端引入活性赖氨酸。 我们发现我们可以很容易地控制病毒体共轭的程度，并且证实了引入的赖氨酸的化学计量生物素化。 为了表征这个用于显示异源蛋白质的模块化平台，我们将模型抗原（链霉亲和素（SA） - 绿色荧光蛋白（GFP），从植物表达和纯化）结合到TMV的表面，产生GFP-SA修饰的病毒颗粒 。 通过使复合物进行氨基酸分析并解决将每个残基的pmol与复合组分的已知氨基酸组成相关的线性方程族进行快速和定量的测定。 我们获得了26％的GFP-SA四聚体负载量，这对应于每个完整病毒粒子显示约2200个GFP部分。 我们评估了GFP装饰病毒粒子在小鼠和豚鼠中的免疫原性，并且相对于未结合的GFP-SA四聚体，在两种物种中发现增强的体液IgG滴度。 在小鼠中，我们在仅用TMV蛋白复合物进行单次免疫后观察到可检测的体液免疫应答。 通过展示整个蛋白质的表现，本研究扩展了TMV作为疫苗支架的效用，超过了通过遗传操作可能的疫苗支架。

    METHOD FOR SCREENING FOR MUTANT VIRUS MOVEMENT PROTEINS (MP)
    5.
    发明申请
    METHOD FOR SCREENING FOR MUTANT VIRUS MOVEMENT PROTEINS (MP) 审中-公开
    用于筛选突变型病毒运动蛋白的方法（MP）

    公开(公告)号：WO2004023137A3

    公开(公告)日：2004-05-06

    申请号：PCT/EP0309795

    申请日：2003-09-04

    Applicant: ARC SEIBERSDORF RES GMBH ,  WAIGMANN ELISABETH

    Inventor： WAIGMANN ELISABETH

    IPC: C07K14/08 ,  C12N15/82 ,  G01N33/50 ,  C07K14/005

    CPC classification number: C07K14/005 ,  C12N15/8283 ,  C12N2770/00022 ,  G01N33/5097

    Abstract: A method for screening for mutant virus movement proteins (MP) which confer virus resistance to a plant transfected with the mutant MP is provided comprising the steps of (a) carrying out at least one point mutation at a regulatory phosphorylation site of the virus MP, (b) transfecting said mutant MP into at least one plant cell of a cellular structure, (c) analysing an intracellular targeting profile and translocation rate of the mutant MP in the cellular structure and (d) comparing the targeting profile and translocation rate of the mutant MP with the targeting profile and translocation rate of a corresponding wild-type MP, whereby is a mutant virus MP which confers virus resistance to a plant shows localisation to movement relevant cellular structures and a lower translation rate.

    Abstract translation: 提供一种筛选突变型病毒运动蛋白（MP）的方法，所述突变体病毒运动蛋白（MP）赋予对具有突变MP的转染植物的病毒抗性，包括以下步骤：（a）在病毒MP的调节性磷酸化位点进行至少一个点突变， （b）将所述突变体MP转染到细胞结构的至少一个植物细胞中，（c）分析细胞结构中突变体MP的细胞内靶向谱和易位率，以及（d）比较所述突变体MP的靶向谱和易位率 具有相应野生型MP的靶向特征和易位速率的突变体MP，其中赋予植物病毒抗性的突变病毒MP显示移动相关细胞结构的定位和较低的翻译率。

    PRODUCTION OF FOREIGN POLYPEPTIDES IN PLANTS AS VIRAL COAT PROTEIN FUSIONS
    7.
    发明申请
    PRODUCTION OF FOREIGN POLYPEPTIDES IN PLANTS AS VIRAL COAT PROTEIN FUSIONS 审中-公开
    在植物中生产外源性多糖作为病毒蛋白融合蛋白

    公开(公告)号：WO01066778A3

    公开(公告)日：2002-05-10

    申请号：PCT/US2001/007355

    申请日：2001-03-08

    IPC: A01H5/00 ,  A61K39/00 ,  C07K14/08 ,  C07K14/445 ,  C12N5/10 ,  C12N7/00 ,  C12N7/01 ,  C12N7/02 ,  C12N15/09 ,  C12N15/40 ,  C12N15/82 ,  C12P21/02 ,  C12P21/04 ,  C12N5/14 ,  C12N15/62

    CPC classification number: C07K14/005 ,  A61K39/00 ,  C07K14/445 ,  C07K2319/00 ,  C12N15/8203 ,  C12N15/8257 ,  C12N15/8258 ,  C12N2770/00022

    Abstract: The present invention relates to foreign peptide sequences fused to the amino-terminal of plant viral structural proteins and a method of their production. Fusion proteins are economically synthesized in plants at high levels by biologically contained tobamoviruses. The foreign peptide sequences can be cleaved from the fusion proteins by proteolytic enzymes or chemical reagents. The foreign peptide sequences of the invention have many uses. Such uses include use as antigens for inducing the production of antibodies having desired binding properties, e.g., protective antibodies, for use as vaccine antigens for the induction of protective immunity, including immunity against parasitic infections, for use as a protein involved in hormonal activity, or for use as a protein involved in immunoregulatory activity.

    Abstract translation: 本发明涉及与植物病毒结构蛋白的氨基末端融合的外来肽序列及其制备方法。 融合蛋白在生物学含有的烟草花叶病毒的高水平植物中经济合成。 外源肽序列可以通过蛋白水解酶或化学试剂从融合蛋白中切割出来。 本发明的外源肽序列具有许多用途。 这样的用途包括用作诱导产生具有所需结合特性的抗体的抗原，例如保护性抗体，用作诱导保护性免疫的疫苗抗原，包括针对寄生虫感染的免疫，用作参与激素活性的蛋白质， 或用作参与免疫调节活性的蛋白质。

    METHOD FOR INDUCING VIRAL RESISTANCE INTO A PLANT
    8.
    发明申请
    METHOD FOR INDUCING VIRAL RESISTANCE INTO A PLANT 审中-公开
    诱导植物体内抗病毒的方法

    公开(公告)号：WO00055301A3

    公开(公告)日：2001-01-25

    申请号：PCT/EP2000/002176

    申请日：2000-03-07

    IPC: A01H5/00 ,  C07K14/08 ,  C12N15/40 ,  C12N15/82

    CPC classification number: C07K14/005 ,  C12N15/8283 ,  C12N2770/00022

    Abstract: The present invention concerns a method for inducing resistance to a virus comprising a TGB2 sequence into a cell plant or a plant, comprising the following steps: preparing a nucleotide construct comprising a nucleotide sequence corresponding to at least 70 % of the nucleotide sequence of TGB2 of said virus or its complementary cDNA, being operably linked to one or more regulatory sequence(s) active in a plant, transforming a plant cell with the nucleotide construct, and possibly regenerating a transgenic plant from the transformed plant cell. The present invention is also related to the plant obtained.

    Abstract translation: 本发明涉及一种诱导对包含TGB2序列的病毒对细胞植物或植物的抗性的方法，包括以下步骤：制备包含对应于TGB2的核苷酸序列的至少70％的核苷酸序列的核苷酸序列 所述病毒或其互补cDNA可操作地连接到在植物中活性的一种或多种调节序列，用所述核苷酸构建体转化植物细胞，以及可能从转化的植物细胞再生转基因植物。 本发明还涉及获得的植物。

    METHOD OF GENETIC MODIFICATION OF A WILD TYPE VIRAL SEQUENCE
    9.
    发明申请
    METHOD OF GENETIC MODIFICATION OF A WILD TYPE VIRAL SEQUENCE 审中-公开
    一种野生型病毒序列的遗传修饰方法

    公开(公告)号：WO00003025A2

    公开(公告)日：2000-01-20

    申请号：PCT/BE1999/000089

    申请日：1999-07-09

    IPC: A01H5/00 ,  C07K14/08 ,  C12N5/10 ,  C12N15/82 ,  C12Q1/68

    CPC classification number: C07K14/005 ,  C12N15/8283 ,  C12N2770/00022

    Abstract: The present invention concerns a method of genetic modification of a TGB-3 wild type viral sequence for reducing or suppressing the possible deleterious effects of the agronomic properties of a transformed plant or plant cell by said TGB-3 viral sequence, comprising the following successive steps: submitting said sequence to point mutation(s) which allow the substitution of at least one amino-acid into a different amino-acid; selecting genetically modified TGB-3 wild type viral sequences having said point mutation(s) and which are not able to promote cell-to-cell movement of a mutant virus having a dysfunctional TGB-3 wild type viral sequence, when expressed in trans from a replicon; further selecting among said genetically modified TGB-3 viral sequences, the specifically genetically modified sequence which inhibits infection with a co-inoculated wild type virus when the mutant form was expressed from a replicon; and recovering said specifically genetically modified TGB-3 viral sequence.

    Abstract translation: 本发明涉及一种TGB-3野生型病毒序列的遗传修饰方法，用于降低或抑制所述TGB-3病毒序列对转化的植物或植物细胞的农学性质的可能有害影响，包括以下连续步骤 ：将所述序列提交至允许将至少一个氨基酸置换成不同氨基酸的点突变; 选择具有所述点突变的遗传修饰的TGB-3野生型病毒序列，并且不能促进具有功能异常的TGB-3野生型病毒序列的突变病毒的细胞间移动，当以 复制品 所述遗传修饰的TGB-3病毒序列进一步选择，当突变形式由复制子表达时，特异性遗传修饰的序列可抑制共同接种的野生型病毒感染; 并回收所述特异性遗传修饰的TGB-3病毒序列。

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