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1. 发明申请

WO2014011853A2 IGY FROM NOROVIRUS P PARTICLES AND THEIR DERIVATIVES 审中-公开
标题翻译： IGY从NOROVIRUS P粒子及其衍生物
公开(公告)号：WO2014011853A2
公开(公告)日：2014-01-16
申请号：PCT/US2013/050044
申请日：2013-07-11
申请人： CHILDREN'S HOSPITAL MEDICAL CENTER , DAI, Ying-Chun
发明人： DAI, Ying-Chun , JIANG, Xi
CPC分类号： C07K16/02 , A61K2039/505 , C07K16/10
摘要： A method for large-scale production of anti-NoV antibodies for use as a potential treatment for NoV disease using passive immunization. NoV-specific immunoglobulins (IgY) can be produced by immunizing chickens with NoV P particles. The birds continuously produced high titers of antibodies in their eggs for at least 3 months, in which NoV-specific antibody levels reached 4.7-9.2 mg/egg yolk. The egg yolk antibodies strongly reacted with NoV P particles by both ELISA and Western blot and blocked NoV virus-like particle (VLP) and P particle binding to the histo-blood group antigen (HBGA) receptors with a BT 50 of about 1 :800. The chicken IgY remain stable at 70°C for 30 min or treatment at pH 4 to 9 for 3 hr, demonstrating that chicken IgY provides large-scale production of anti-NoV antibodies for use in passive immunization against NoV infection.
摘要翻译：用于大规模生产抗NoV抗体的方法用作使用被动免疫的NoV疾病的潜在治疗。 NoV特异性免疫球蛋白（IgY）可以通过用NoV P粒子免疫鸡来产生。鸡在鸡蛋中持续产生高滴度的抗体至少3个月，其中NoV特异性抗体水平达到4.7-9.2mg /蛋黄。蛋黄抗体通过ELISA和Western印迹与NoV P颗粒强烈反应，并且用BT 50蛋白质阻断了NoV病毒样颗粒（VLP）和P颗粒与组织血型抗原（HBGA）大约1：800。鸡IgY在70℃保持稳定30分钟或在pH 4至9处理3小时，表明鸡IgY提供大规模生产的抗NoV抗体用于针对NoV感染的被动免疫。

2. 发明申请

WO2010144602A2 ANTIGEN-NOROVIRUS P-DOMAIN MONOMERS AND DIMERS, ANTIGEN-NOROVIRUS P-PARTICLE MOLECULES, AND METHODS FOR THEIR MAKING AND USE 审中-公开
标题翻译：抗逆转录病毒单核细胞和二聚体，抗逆转录病毒颗粒分子及其制备和使用方法
公开(公告)号：WO2010144602A2
公开(公告)日：2010-12-16
申请号：PCT/US2010/038008
申请日：2010-06-09
申请人： CHILDREN'S HOSPITAL MEDICAL CENTER , JIANG, Xi , TAN, Ming
发明人： JIANG, Xi , TAN, Ming
IPC分类号： C07K19/00 , C12N15/63 , A61K39/295 , A61P37/00
CPC分类号： C07K7/06 , A61K38/00 , A61K39/12 , A61K39/125 , A61K2039/5258 , A61K2039/542 , A61K2039/543 , A61K2039/55566 , A61K2039/58 , A61K2039/6075 , A61K2039/645 , A61K2039/70 , C07K5/10 , C12N2720/12334 , C12N2770/16034
摘要： A substituted Norovirus capsid protein monomer, having only the P-domain and called an antigen-Norovirus P-domain monomer, includes a foreign antigen inserted into one or more of three surface loops present on each P-domain monomer by molecular cloning. The antigen-P-domain monomer can assemble spontaneously into an octahedral form, called an antigen-Norovirus P-particle, that is composed of 24 copies of the antigen-P-domain monomer. Each substituted P-domain monomer will contain one to three copies of the foreign antigen, for a total of 24-72 antigen copies on each antigen-P-particle. The antigen-P-particle is useful in methods for diagnosing, immunizing and treating individuals infected with a foreign virus, for example Rotavirus, and can serve as a carrier for presentation of foreign antigens for development of novel vaccines against many infectious and non-infectious diseases. The substituted Norovirus P-particles can be readily produced in E. coli and yeast, are highly stable and tolerate a wide range of physio-chemical conditions. A modified Norovirus P-domain monomer includes one or more restriction recognition sites inserted within one or more of the three loops of the P-domain monomers, to provide user-friendly cloning cassettes for conveniently inserting candidate foreign antigens into the surface loops. The P-particle-VP8 chimeras may also serve as a dual vaccine against both rotavirus and norovirus.
摘要翻译：仅具有P-结构域并称为抗原 - 诺如病毒P-结构域单体的取代的诺如病毒衣壳蛋白单体包括通过分子克隆插入每个P-结构域单体上存在的三个表面环中的一个或多个的外源抗原。抗原-P结构域单体可以自发组装成称为抗原 - 诺如病毒P颗粒的八面体形式，其由24个抗原-P结构域单体组成。每个取代的P-结构域单体将含有一至三个拷贝的外源抗原，每个抗原-P-粒子上共有24-72个抗原拷贝。抗原-P颗粒可用于诊断，免疫和治疗感染外来病毒的个体的方法，例如轮状病毒（Rotavirus），并且可以用作外源抗原的载体，用于开发针对许多感染性和非感染性的新型疫苗疾病。取代的诺如病毒P颗粒可以容易地在大肠杆菌和酵母中产生，高度稳定并且耐受广泛的生理化学条件。修饰的诺如病毒P结构域单体包括插入在P结构域单体的三个环中的一个或多个环内的一个或多个限制性识别位点，以提供方便地将候选外源抗原插入到表面环中的用户友好的克隆盒。 P粒子VP8嵌合体也可以用作针对轮状病毒和诺如病毒的双重疫苗。

3. 发明申请

WO2006138514A2 NOVEL NOROVIRUS PARTICLE FOR USE AS AN ANTIVIRAL OR VACCINE 审中-公开
标题翻译：用作抗病毒剂或疫苗的新型尿毒症颗粒
公开(公告)号：WO2006138514A2
公开(公告)日：2006-12-28
申请号：PCT/US2006/023407
申请日：2006-06-15
申请人： CHILDREN'S HOSPITAL MEDICAL CENTER , JIANG, Xi , TAN, Ming
发明人： JIANG, Xi , TAN, Ming
IPC分类号： A61K39/12
CPC分类号： G01N33/56983 , A61K39/00 , A61K2039/5258 , C07K14/005 , C12N2770/16022 , C12N2770/16023 , Y02A50/472 , Y02A50/52
摘要： Norovirus capsid protein monomers having only the P domain, and not the hinge or S domain, can assemble spontaneously into an icosahedral form herein called the P-particle. Factors affecting the formation and stability of the P-particle, as well as providing methods for diagnosing and treating Norovirus-infected individuals and creating a vaccine for prevention of Norovirus infection are presented.
摘要翻译：仅具有P结构域而不是铰链或S结构域的诺如病毒衣壳蛋白单体可以自发地组装成本文称为P颗粒的二十面体形式。提出了影响P颗粒形成和稳定性的因素，以及提供诊断和治疗诺如病毒感染个体以及创造用于预防诺如病毒感染的疫苗的方法。

4. 发明申请

WO2014011829A1 PROTEIN COMPLEX SYSTEM FOR INCREASED IMMUNOGENICITY AND FUNCTIONALITY, AND METHODS MAKING AND USE 审中-公开
标题翻译：增强免疫原性和功能的蛋白复合体系及其制备和使用方法
公开(公告)号：WO2014011829A1
公开(公告)日：2014-01-16
申请号：PCT/US2013/050004
申请日：2013-07-11
申请人： CHILDREN'S HOSPITAL MEDICAL CENTER
发明人： JIANG, Xi , TAN, Ming , WANG, Leyi
IPC分类号： C07K19/00 , C07K14/005 , C12N15/62 , A61K39/295
CPC分类号： C07K14/005 , A61K39/12 , A61K39/385 , A61K2039/543 , A61K2039/627 , A61K2039/64 , A61K2039/70 , C07K2319/23 , C07K2319/40 , C07K2319/50 , C07K2319/70 , C12N2720/12322 , C12N2720/12334 , C12N2760/16022 , C12N2760/16034 , C12N2760/16134 , C12N2770/16022 , C12N2770/16034 , C12N2770/28122
摘要： Genes for proteins which spontaneously form dimers and/or oligomers can be recombinantly linked together, which upon expression in E. coli produces stable dimeric fusion proteins that spontaneously self-assemble into enormous, polyvalent complexes having increased immunogenicity and functionality. Linear, network and agglomerate complexes with enormous sizes and polyvalences are constructed using glutathione S-transferase, Norovirus P domains (NoV P - and NoV + ), the protruding (P) domain of hepatitis E virus (HEV P), the astrovirus P domain (AstV), a monomeric peptide epitope (M2e of influenza virus), and/or a protein antigen (VP8* of rotavirus) fused in different combinations. The resulting complexes can contain hundreds to thousands NoV P-protein, HEV, AstV, M2e and/or VP8* copies and exhibit higher immunogenicity than the individual proteins alone. The large size and multivalent nature of the complexes are candidates as a bivalent or multivalent vaccines against Norovirus and other pathogens, and for generation of antibodies for diagnosis and research purposes.
摘要翻译：自发形成二聚体和/或寡聚体的蛋白质的基因可以重组连接在一起，其在大肠杆菌中表达后产生稳定的二聚融合蛋白，其自发地自组装成具有增加的免疫原性和功能性的巨大的多价复合物。使用谷胱甘肽S-转移酶，诺如病毒P结构域（NoV P-和NoV +），戊型肝炎病毒（HEV P）的突出（P）结构域，星状病毒P结构域（...）构建具有巨大尺寸和多价性的线性，网络和聚集体复合物 AstV），单体肽表位（流感病毒的M2e）和/或以不同组合融合的蛋白质抗原（轮状病毒的VP8 *）。所得复合物可以含有数百至数千个NoV P蛋白，HEV，AstV，M2e和/或VP8 *拷贝，并且比单独的蛋白质具有更高的免疫原性。复合物的大尺寸和多价性质是针对诺如病毒和其他病原体的二价或多价疫苗的候选物，以及为诊断和研究目的产生抗体。

5. 发明申请

WO2003101176A3 METHOD, COMPOSITION AND KIT FOR ANTIGENIC BINDING OF NORWALK-LIKE VIRUSES 审中-公开
公开(公告)号：WO2003101176A3
公开(公告)日：2003-12-11
申请号：PCT/US2003/017247
申请日：2003-06-02
申请人： CHILDREN'S HOSPITAL MEDICAL CENTER , LE PENDU, Jacques , JIANG, Xi
发明人： LE PENDU, Jacques , JIANG, Xi
IPC分类号： C12Q1/70
摘要： A method for detecting a Norwalk-Like Virus (NLV) in a biological sample, comprising the steps of: obtaining a biological sample suspected of containing a NLV; contacting the biological sample with at least one human histo-blood group antigen to allow formation of a complex of the NLV with the antigen; and detecting the antigen NLV complex. The antigen-NLV complex can be detected by contacting the NLV-antigen complex with a NLV antibody that binds at an epitope of the NLV, and detecting the NLV antibody. The invention also includes a method for identifying compounds, and the compounds, that can inhibit the binding between a Norwalk-Like Virus (NLV) and a histo-blood group antigen. The method includes the steps of contacting the NLV target with a compound; subsequently contacting the NLV with a standard compound that is known to be bound at a determinant binding site of the NLV; and determining whether the binding of the standard compound is decreased in the presence of the test compound, the decrease in binding being an indication that the test compound inhibits the binding activity of the NLV with the standard compound. In preferred embodiments, the standard compound is a histo-blood group antigen.

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