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1. 发明授权

US08554493B2 Viral polyhedra complexes and methods of use 有权
标题翻译：病毒多面体复合物及其使用方法
公开(公告)号：US08554493B2
公开(公告)日：2013-10-08
申请号：US12529110
申请日：2008-02-28
申请人： Peter Metcalf , Fasseli Joseph Coulibaly , Hajime Mori , Norio Hamada , Keiko Ikeda , Yui Lam Elaine Chiu , Hiroshi Ijiri
发明人： Peter Metcalf , Fasseli Joseph Coulibaly , Hajime Mori , Norio Hamada , Keiko Ikeda , Yui Lam Elaine Chiu , Hiroshi Ijiri
IPC分类号： G06F19/00
CPC分类号： C07K14/005 , A01N25/34 , A01N63/00 , A61K47/6901 , A61K47/6949 , B82Y5/00 , C07K14/475 , C07K14/485 , C07K14/50 , C07K2319/735 , C12N2710/14043 , C12N2720/12022 , A01N61/00 , A01N63/02 , A01N2300/00
摘要： Cypoviruses and baculoviruses are notoriously difficult to eradicate because the virus particles are embedded in micron-sized protein crystals called polyhedra. The remarkable stability of polyhedra means that like bacterial spores these insect viruses remain infectious for years in soil. Although these unique in vivo protein crystals have been extensively characterized since the early 1900s, their atomic organization remains elusive. Here we describe the 2 crystal structure of both recombinant and infectious silkworm cypovirus polyhedra determined using 5-12 micron crystals purified from insect cells. These are the smallest crystals yet used for de novo X-ray protein structure determination. It was found that polyhedra are made of trimers of the viral polyhedrin protein and contain nucleotides. Although the shape of these building blocks is reminiscent of some capsid trimers, polyhedrin has a new fold and has evolved to assemble in vivo into 3-D cubic crystals rather than icosahedral shells. The polyhedrin trimers are extensively cross-linked in polyhedra by non-covalent interactions and pack with an exquisite molecular complementarity similar to that of antigen-antibody complexes. The resulting ultra-stable and sealed crystals shield the virus particles from environmental damage. The structure suggests that polyhedra can serve as the basis for the development of robust and versatile nanoparticles for biotechnological applications such as in cell culture systems, microarrays and biopesticides.
摘要翻译：由于病毒颗粒嵌入称为多面体的微米级蛋白质晶体中，Cypoviruses和杆状病毒众所周知难以根除。多面体的显着稳定意味着像细菌孢子一样，这些昆虫病毒在土壤中仍然具有传染性多年。虽然这些独特的体内蛋白质晶体自20世纪初以来已被广泛地表征，但它们的原子组织仍然难以捉摸。这里我们描述使用从昆虫细胞纯化的5-12微米晶体测定的重组和感染性蚕病毒多面体的2晶体结构。这些是用于从头X射线蛋白质结构测定的最小晶体。发现多面体由病毒多角体蛋白的三聚体构成并含有核苷酸。尽管这些结构单元的形状让人联想到一些衣壳三聚体，但多角体蛋白具有新的折叠，并已发展成体内装配成3-D立方晶体而不是二十面体壳。多面体三聚体通过非共价相互作用在多面体中广泛交联，并具有类似于抗原 - 抗体复合物的精细分子互补性。所得到的超稳定和密封的晶体屏蔽病毒颗粒免受环境破坏。该结构表明，多面体可以作为开发用于生物技术应用的强壮和多功能纳米颗粒的基础，例如在细胞培养系统，微阵列和生物杀虫剂中。

2. 发明申请

US20100216651A1 VIRAL POLYHEDRA COMPLEXES AND METHODS OF USE 有权
标题翻译： VIRAL POLYHEDRA复合物及其使用方法
公开(公告)号：US20100216651A1
公开(公告)日：2010-08-26
申请号：US12529110
申请日：2008-02-28
申请人： Peter Metcalf , Fasseli Joseph Coulibaly , Hajime Mori , Norio Hamada , Keiko Ikeda , Yui Lam Elaine Chiu , Hiroshi Ijiri
发明人： Peter Metcalf , Fasseli Joseph Coulibaly , Hajime Mori , Norio Hamada , Keiko Ikeda , Yui Lam Elaine Chiu , Hiroshi Ijiri
IPC分类号： C40B30/00 , C12N7/00 , C12N5/071
CPC分类号： C07K14/005 , A01N25/34 , A01N63/00 , A61K47/6901 , A61K47/6949 , B82Y5/00 , C07K14/475 , C07K14/485 , C07K14/50 , C07K2319/735 , C12N2710/14043 , C12N2720/12022 , A01N61/00 , A01N63/02 , A01N2300/00
摘要： Cypoviruses and baculoviruses are notoriously difficult to eradicate because the virus particles are embedded in micron-sized protein crystals called polyhedra. The remarkable stability of polyhedra means that like bacterial spores these insect viruses remain infectious for years in soil. Although these unique in vivo protein crystals have been extensively characterized since the early 1900s, their atomic organization remains elusive. Here we describe the 2 crystal structure of both recombinant and infectious silkworm cypovirus polyhedra determined using 5-12 micron crystals purified from insect cells. These are the smallest crystals yet used for de novo X-ray protein structure determination. It was found that polyhedra are made of trimers of the viral polyhedrin protein and contain nucleotides. Although the shape of these building blocks is reminiscent of some capsid trimers, polyhedrin has a new fold and has evolved to assemble in vivo into 3-D cubic crystals rather than icosahedral shells. The polyhedrin trimers are extensively cross-linked in polyhedra by non-covalent interactions and pack with an exquisite molecular complementarity similar to that of antigen-antibody complexes. The resulting ultra-stable and sealed crystals shield the virus particles from environmental damage. The structure suggests that polyhedra can serve as the basis for the development of robust and versatile nanoparticles for biotechnological applications such as in cell culture systems, microarrays and biopesticides.
摘要翻译：由于病毒颗粒嵌入称为多面体的微米级蛋白质晶体中，Cypoviruses和杆状病毒众所周知难以根除。多面体的显着稳定意味着像细菌孢子一样，这些昆虫病毒在土壤中仍然具有传染性多年。虽然这些独特的体内蛋白质晶体自20世纪初以来已被广泛地表征，但它们的原子组织仍然难以捉摸。这里我们描述使用从昆虫细胞纯化的5-12微米晶体测定的重组和感染性蚕病毒多面体的2晶体结构。这些是用于从头X射线蛋白质结构测定的最小晶体。发现多面体由病毒多角体蛋白的三聚体构成并含有核苷酸。尽管这些结构单元的形状让人联想到一些衣壳三聚体，但多角体蛋白具有新的折叠，并已发展成体内装配成3-D立方晶体而不是二十面体壳。多面体三聚体通过非共价相互作用在多面体中广泛交联，并具有类似于抗原 - 抗体复合物的精细分子互补性。所得到的超稳定和密封的晶体屏蔽病毒颗粒免受环境破坏。该结构表明，多面体可以作为开发用于生物技术应用的强壮和多功能纳米颗粒的基础，例如在细胞培养系统，微阵列和生物杀虫剂中。

3. 发明授权

US07432347B2 Cytoplasmic polyhedrosis virus polyhedrin protein complex 有权
标题翻译：细胞质多角体病毒多角体蛋白复合物
公开(公告)号：US07432347B2
公开(公告)日：2008-10-07
申请号：US10415096
申请日：2001-10-30
申请人： Yoshimoto Ohta , Hajime Mori , Keiko Ikeda
发明人： Yoshimoto Ohta , Hajime Mori , Keiko Ikeda
IPC分类号： C07K14/00 , C12P21/06 , C12P21/04 , C12N7/00 , C12N5/06
CPC分类号： C07K14/005 , C12N15/8257 , C12N15/86 , C12N2710/14143 , C12N2720/12022 , C12P21/02
摘要： To provide a useful objective protein coated with a protective protein, a protein complex that is produced in cells and has a structure that the objective protein is occluded with the viral occlusion body protein. A protein complex having the structure that the objective protein is occluded with the viral occlusion body protein, produced in insect cells or plant cells, preferably produced by incorporation of the objective protein in the crystalline form during crystallization of polyhedron derived from polyhedrosis virus, preferably a protein complex in which the polyhedrin contributes to improvement of stability, or protection, or improvement of preservability, or combination thereof, of the objective protein.
摘要翻译：提供涂覆有保护性蛋白质的有用的目标蛋白质，其是在细胞中产生并具有目标蛋白质被病毒闭塞体蛋白质封闭的结构的蛋白质复合物。具有在昆虫细胞或植物细胞中产生的目的蛋白被病毒闭塞体蛋白封闭的结构的蛋白质复合物，优选通过在衍生自多角体病毒的多面体结晶过程中结晶形成目标蛋白质而产生，优选为蛋白质复合物，其中多角体蛋白有助于提高目标蛋白质的稳定性或保护性，或其保存性或其组合的改善。

4. 发明授权

US07619060B2 Cytoplasmic polyhedrosis virus protein complex of a polyhedrin and a VP3 polypeptide 有权
标题翻译：多角体蛋白和VP3多肽的细胞质多角体病毒蛋白复合物
公开(公告)号：US07619060B2
公开(公告)日：2009-11-17
申请号：US10541752
申请日：2004-01-07
申请人： Keiko Ikeda
发明人： Keiko Ikeda
IPC分类号： C07K14/00 , C12P21/06 , C12P21/04 , C12N7/00 , C12N5/06
CPC分类号： G01N33/54393 , C07K2319/00
摘要： It is intended to provide a protein complex and a production process whereby the protein complex can be efficiently produced without lowering its function. It is also intended to provide use of the protein complex in a biosensor, an immobilized enzyme and so on. A protein complex comprising a polyhedral protein having an insect virus encapsulated therein and a target protein having a restricted region of a capsid protein VP3 of cytoplasmic polyhedrosis virus, more specifically, a region which is either a region from the N-terminus to the 40th amino acid residue or a region from the 41st amino acid residue to the 79th amino acid residue as an embedding signal for polyhedron, and a process for producing the same. The polyhedral protein has an effect on improvement in the stability of the target protein, protection thereof or improvement in the preservation properties thereof, or a combination thereof.
摘要翻译：旨在提供蛋白质复合物和生产方法，由此可以有效地制备蛋白质复合物而不降低其功能。还旨在提供生物传感器，固定化酶等中的蛋白质复合物的使用。包含其中包含昆虫病毒的多面体蛋白质的蛋白质复合物和具有细胞质多角体病毒的衣壳蛋白VP3的限制区域的靶蛋白质，更具体地说，是从N末端到第40位的区域酸残基或第41位氨基酸残基至第79位氨基酸残基的区域，作为多面体的嵌入信号及其制造方法。多面体蛋白质可以改善靶蛋白的稳定性，其保护作用，或其保存性能的提高，或其组合。

5. 发明申请

US20060155114A1 Protein complex, process for producing the same and use thereof 有权
标题翻译：蛋白质复合物，其制备方法及其应用
公开(公告)号：US20060155114A1
公开(公告)日：2006-07-13
申请号：US10541752
申请日：2004-01-07
申请人： Keiko Ikeda
发明人： Keiko Ikeda
IPC分类号： C07K14/005 , C12Q1/70
CPC分类号： G01N33/54393 , C07K2319/00
摘要： It is intended to provide a protein complex and a production process whereby the protein complex can be efficiently produced without lowering its function. It is also intended to provide use of the protein complex in a biosensor, an immobilized enzyme and so on. A protein complex comprising a polyhedral protein having an insect virus encapsulated therein and a target protein having a restricted region of a capsid-protein VP3 of cytoplasmic polyhedrosis virus, more specifically, a region which is either a region from the N-terminus to the 40th amino acid residue or a region from the 41st amino acid residue to the 79th amino acid residue as an embedding signal for polyhedron, and a process for producing the same. The polyhedral protein has an effect on improvement in the stability of the target protein, protection thereof or improvement in the preservation properties thereof, or a combination of any of these. The target protein is at least one member selected from the group consisting of fluorescent or light-emitting proteins, enzymes, antigens, antibodies, cytokines, receptors and bioactive proteins. A biosensor characterized in that the above-described protein complex is arranged in dots or lines on a substrate and immobilized thereon.
摘要翻译：旨在提供蛋白质复合物和生产方法，由此可以有效地制备蛋白质复合物而不降低其功能。还旨在提供生物传感器，固定化酶等中的蛋白质复合物的使用。包含其中包含昆虫病毒的多面体蛋白质的蛋白质复合物和具有细胞质多角体病毒的衣壳蛋白VP3的限制区域的靶蛋白质，更具体地说，是从N末端至第40位的区域氨基酸残基或第41位氨基酸残基至第79位氨基酸残基的区域，作为多面体的嵌入信号及其制造方法。多面体蛋白质对于提高目标蛋白质的稳定性，保护性，或者其保存性能的提高，或其组合均有影响。靶蛋白是选自荧光或发光蛋白，酶，抗原，抗体，细胞因子，受体和生物活性蛋白质中的至少一种。一种生物传感器，其特征在于，将上述蛋白质复合体以点或线排列在基板上并固定在基板上。

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