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    • 5. 发明申请
    • Rapid sterilization and vaccine preparation
    • 快速灭菌和疫苗制备
    • US20020182107A1
    • 2002-12-05
    • US09924266
    • 2001-08-07
    • James A. Laugharn JR.David W. BradleyRobert A. Hess
    • A61L002/02C12N007/04
    • A61L2/02A23L3/0155A61L2/0011
    • The invention is based on the discovery that biological and non-biological materials can be sterilized, decontaminated, or disinfected by repeatedly cycling between relatively high and low pressures. Pressure cycling can be carried out at low, ambient, or elevated temperatures (e.g., from about null40null C. to about 95null C., or intermediate ranges). New methods based on this discovery can have applications in, for example, the preparation of vaccines, the sterilization of blood plasma or serum, plant, animal, and human tissue, sputum, urine, feces, water, and ascites, the decontamination of military devices, food and beverage production, and the disinfection of medical equipment. The new methods can also be incorporated into production processes or research procedures.
    • 本发明基于以下发现:生物和非生物材料可以通过在相对高和低的压力之间重复循环进行灭菌,去污或消毒。 压力循环可以在低,环境或升高的温度(例如,约-40℃至约95℃或中等范围)下进行。 基于该发现的新方法可以应用于例如疫苗的制备,血浆或血清,植物,动物和人体组织,痰液,尿液,粪便,水和腹水的灭菌,军事的去污 设备,食品和饮料生产以及医疗设备的消毒。 新的方法也可以纳入生产过程或研究程序。
    • 6. 发明申请
    • Efficient generation of adenovirus-based libraries by positive selection of adenoviral recombinants through ectopic expression of the adenovirus protease
    • 通过腺病毒蛋白酶的异位表达阳性选择腺病毒重组体来有效地产生基于腺病毒的文库
    • US20020051966A1
    • 2002-05-02
    • US09843949
    • 2001-04-30
    • Bernard MassieSeyyed Mehdy ElahiWahiba Qualikene
    • C12N007/00C12Q001/70G01N033/53C12N015/861C12N007/04C12N015/09C12N015/70C07H021/02C07H021/04C12N015/74
    • C12N7/00A61K39/00A61K48/00C12N15/86C12N2710/10343C12N2710/10352C12N2810/50C12N2830/003C12N2840/20C12N2840/203
    • Disclosed is a new system for generating recombinant adenovirus vectors and adenovirus-based expression libraries, by positive selection of recombinants deleted for the endogenous protease gene, which gene is expressibly cloned into another region of the adenoviral genome. In a preferred embodiment, the invention allows positive selection of E1-deleted, protease-deleted recombinant adenovirus vectors comprising an exogenous gene or an expressible piece of exogenous DNA, by providing an expression cassette comprising the protease gene and the exogenous DNA inserted in place of E1 region in a shuttle vector. In vivo recombination of the shuttle vector with a protease-deleted adenoviral genome in suitable non-complementing cells generates viable recombinants only when rescuing the protease cloned in E1 region. Non-recombinant viral genomes are not able to grow due to the deletion of the protease gene, ensuring that only recombinant viral plaques are generated. This positive selection can be used for the generation of a large number of high purity recombinant adenovirus vectors and allows generation of adenovirus-based libraries with diversity exceeding 106 clones.
    • 公开了一种用于产生重组腺病毒载体和基于腺病毒的表达文库的新系统,通过阳性选择为内源性蛋白酶基因缺失的重组体,该基因可表达克隆到腺病毒基因组的另一区域。 在一个优选的实施方案中,本发明允许通过提供包含蛋白酶基因的表达盒和插入的外源DNA来代替包含外源基因或可表达外源DNA片段的E1缺失的,蛋白酶缺失的重组腺病毒载体,代替 E1区域在穿梭载体中。 穿梭载体与合适的非补体细胞中的蛋白酶缺失型腺病毒基因组的体内重组仅在拯救克隆在E1区域中的蛋白酶时产生可行的重组体。 非重组病毒基因组由于蛋白酶基因的缺失而不能生长,确保仅产生重组病毒噬菌斑。 这种阳性选择可用于产生大量高纯度重组腺病毒载体,并且允许产生具有超过106个克隆的多样性的基于腺病毒的文库。
    • 10. 发明申请
    • Attenuated recombinant rabies virus mutants and live vaccines thereof
    • 减毒重组狂犬病毒突变体及其活疫苗
    • US20020164356A1
    • 2002-11-07
    • US10128628
    • 2002-04-23
    • Teshome Mebatsion
    • A61K048/00A01N063/00A61K039/12C12N007/04
    • C12N7/00A61K39/12A61K39/205A61K2039/525A61K2039/5254A61K2039/542C07K14/005C12N2760/20122C12N2760/20134C12N2760/20161
    • The present invention describes recombinant RV mutants comprising a combined mutation in two different parts of the viral genome, involving the P and the G genes. The mutations in the P gene preferably encompass residues 139 to 170, more preferably residues 139 to 149, most preferably residues 143-149. The mutation can be a substitution or deletion of one or more amino acids in the above region, as well as combinations of deletion and substitution. Preferred mutants according to the invention may be obtained by deleting residues 143 to 149 or 139 to 149 of the phosphoprotein (P) of rabies virus and simultaneously replacing the Arg at position 333 of the glycoprotein into another residue, preferably Asp instead of Arg. Surprisingly, when these mutations were introduced into rabies viruses lacking Arg at position 333 of their G protein, a dramatic reduction in pathogenicity for suckling mice was observed. This unexpected finding has a profound advantage in developing more safe live attenuated rabies vaccines. The mutation in the G gene may comprise a mutation of the Arg333 codon into a codon that differs by one, two or three nucleotides from said Arg333 codon. Preferably the mutants are mutants of a RV strain in which all three nucleotides of the Arg333 codon are substituted.
    • 本发明描述了重组RV突变体,其包含病毒基因组的两个不同部分中的组合突变,涉及P和G基因。 P基因中的突变优选包括残基139至170,更优选残基139至149,最优选残基143-149。 突变可以是上述区域中一个或多个氨基酸的取代或缺失,以及缺失和取代的组合。 根据本发明的优选突变体可以通过删除狂犬病病毒的磷蛋白(P)的143至149或139至149位,同时将糖蛋白333位的Arg同时替换为另一残基,优选Asp而不是Arg来获得。 令人吃惊的是,当这些突变引入到其G蛋白333位缺乏Arg的狂犬病病毒时,观察到乳鼠的致病性显着降低。 这种意想不到的发现在开发更安全的减毒狂犬病疫苗方面具有深远的优势。 G基因中的突变可以包括Arg333密码子的突变成从所述Arg333密码子不同一个,两个或三个核苷酸的密码子。 优选地,突变体是其中Arg333密码子的所有三个核苷酸都被取代的RV菌株的突变体。