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    Outer membrane protein P1 and peptides of Haemophilus influenzae type B
    1.
    发明授权
    Outer membrane protein P1 and peptides of Haemophilus influenzae type B 失效
    外膜蛋白P1和B型流感嗜血杆菌肽

    公开(公告)号：US5985288A

    公开(公告)日：1999-11-16

    申请号：US472172

    申请日：1995-06-07

    申请人： Robert S. Munson, Jr. ,  Susan Grass ,  Pele Chong ,  Yan-Ping Yang ,  Raafat Fahim ,  Dwo Yan Charles Sia ,  Patrick McVerry ,  Michel Klein

    发明人： Robert S. Munson, Jr. ,  Susan Grass ,  Pele Chong ,  Yan-Ping Yang ,  Raafat Fahim ,  Dwo Yan Charles Sia ,  Patrick McVerry ,  Michel Klein

    IPC分类号： A61K39/00 ,  A61K39/102 ,  A61K39/385 ,  A61P31/04 ,  A61P31/12 ,  A61P31/16 ,  C07K14/005 ,  C07K14/195 ,  C07K14/285 ,  C07K14/41 ,  C07K14/705 ,  C07K19/00 ,  C12N15/09 ,  C12N15/31 ,  C12P21/02 ,  C12R1/19 ,  C12R1/21 ,  G01N33/569

    CPC分类号： C07K14/285 ,  A61K39/00

    摘要： The gene for outer membrane protein P1 of Haemophilus influenzae b is expressed in E. coli. Methods for expression and demonstration of the immunogenicity of recombinant P1 and portions thereof are disclosed, along with an improved method for the purification of P1. The nucleotide sequence of the P1 gene and the derived amino acid sequence of the P1 protein of Haemophilus influenzae type b are disclosed and the methods used to determine the same. Also disclosed are the methods used to clone and express the P1 gene as well as the purification protocol for the P1 gene products (recombinant P1 and P1 fusion proteins). Fourteen peptides are synthesized corresponding to specific sequences of the mature P1 protein. The use of the P1 protein as n immunogens for immunization against the disease caused by Haemophilus influenzae type b and the use of the protein as a carrier for conjugation with an oligosaccharide derived from Haemophilus to generate a potentially efficacious vaccine against the disease, are described. Also disclosed is the use of P1 peptide-conjugates as immunizing agents to elicit anti-Haemophilus influenzae type b antibodies.

    摘要翻译： 流感嗜血杆菌b的外膜蛋白P1的基因在大肠杆菌中表达。 公开了用于表达和证明重组P1及其部分的免疫原性的方法以及用于纯化P1的改进方法。 公开了P1型基因的核苷酸序列和B型流感嗜血杆菌的P1蛋白的衍生氨基酸序列，并且用于确定它们的方法。 还公开了用于克隆和表达P1基因的方法以及P1基因产物（重组P1和P1融合蛋白）的纯化方案。 对应于成熟P1蛋白的特异性序列合成14个肽。 描述了使用P1蛋白作为免疫原免疫针对由流感嗜血杆菌b型引起的疾病的免疫，以及使用蛋白质作为与源自嗜血杆菌的寡糖缀合的载体以产生针对该疾病的潜在有效疫苗。 还公开了使用P1肽缀合物作为免疫剂以引发抗流感嗜血杆菌b型抗体。

    Membrane proteins and peptides of haemophilus influenzae type B
    2.
    发明授权
    Membrane proteins and peptides of haemophilus influenzae type B 失效
    乙型流感嗜血杆菌膜蛋白和肽

    公开(公告)号：US5916562A

    公开(公告)日：1999-06-29

    申请号：US123245

    申请日：1993-09-20

    申请人： Robert S. Munson, Jr. ,  Robert Tolan ,  Pele Chong ,  Raafat Fahim ,  Patrick McVerry ,  Michel Klein

    发明人： Robert S. Munson, Jr. ,  Robert Tolan ,  Pele Chong ,  Raafat Fahim ,  Patrick McVerry ,  Michel Klein

    IPC分类号： A61K39/00 ,  A61P31/04 ,  C07K14/285 ,  A61K39/02

    CPC分类号： C07K14/285 ,  A61K39/00

    摘要： The nucleotide sequence of the P2 gene and the derived amino acid sequence of the P2 protein of Haemophilus influenzae type b are disclosed and the methods used to determine the same. Also disclosed are the methods used to clone and express the P2 gene as well as the purification protocol for the gene and protein. Also disclosed is the synthesis of peptides corresponding to the N-terminal and C-terminal ends of the P2 protein. Also disclosed is the use of the P2 protein as a protective agent for the disease caused by Haemophilus influenzae type b and the use of the protein as a carrier for conjugation with an oligosaccharide derived from Haemophilus to generate a potentially efficacious vaccine against the disease. Also disclosed is the use of P2 peptide-conjugates as immunising agents.

    摘要翻译： 公开了P2型基因的核苷酸序列和流感嗜血杆菌b型的P2蛋白的衍生氨基酸序列，并且用于确定其的方法。 还公开了用于克隆和表达P2基因的方法以及用于基因和蛋白质的纯化方案。 还公开了对应于P2蛋白的N末端和C末端的肽的合成。 还公开了P2蛋白作为由流感嗜血杆菌b型引起的疾病的保护剂的用途，以及使用该蛋白质作为与衍生自嗜血杆菌的寡糖缀合的载体以产生针对该疾病的潜在有效的疫苗。 还公开了使用P2肽缀合物作为免疫剂。

    Purification of a pertussis outer membrane protein
    3.
    发明授权
    Purification of a pertussis outer membrane protein 有权

    公开(公告)号：US06444211B1

    公开(公告)日：2002-09-03

    申请号：US09327527

    申请日：1999-06-08

    申请人： Gail Jackson ,  Raafat Fahim ,  Larry Tan ,  Pele Chong ,  John Vose ,  Michel Klein

    发明人： Gail Jackson ,  Raafat Fahim ,  Larry Tan ,  Pele Chong ,  John Vose ,  Michel Klein

    IPC分类号： A61K3910

    摘要： Pertactin (formerly 69 kDa protein) is recovered in stable biologically pure form having no detectable adenylate cyclase activity from fermentation broth from the fermentation of Bordetella pertussis as well as from the cells. The broth is processed to selectively remove pertussis toxin (PT) and filamentous haemagglutinin (FHA), the pertactin is precipitated by ammonium sulphate and the precipitate is dissolved in buffer at pH 6.0 to 8.5, the solution then is passed through hydroxyapatite and ion-exchange chromatograph columns before final ultrafiltration. Cells are extracted with urea and the extract ultrafiltered and diafiltered. The pertactin is precipitated from the extract and the precipitate processed as above. In a variation, the broth is contacted with ammonium sulphate to precipitate pertactin, PT and FHA, the precipitate is dissolved and the PT and FHA selectively removed, before the solution is passed to the chromatograph columns.

    Purification of a pertussis outer membrane protein
    4.
    发明授权
    Purification of a pertussis outer membrane protein 失效
    百日咳外膜蛋白的纯化

    公开(公告)号：US5667787A

    公开(公告)日：1997-09-16

    申请号：US433644

    申请日：1995-05-04

    申请人： Gail Jackson ,  Raafat Fahim ,  Larry Tan ,  Pele Chong ,  John Vose ,  Michel Klein

    发明人： Gail Jackson ,  Raafat Fahim ,  Larry Tan ,  Pele Chong ,  John Vose ,  Michel Klein

    IPC分类号： A61K39/00 ,  A61K39/10 ,  A61P31/04 ,  C07K1/18 ,  C07K1/34 ,  C07K14/005 ,  C07K14/195 ,  C07K14/235 ,  C07K14/41 ,  C12P21/00 ,  C12R1/01 ,  A23J1/00 ,  C07K1/00 ,  C08H1/00

    CPC分类号： C07K14/235 ,  A61K39/00 ,  Y10S530/825

    摘要： Pertactin (formerly 69 kDa protein) is recovered in stable biologically pure form having no detectable adenylate cyclase activity from fermentation broth from the fermentation of Bordetella pertussis as well as from the cells. The broth is processed to selectively remove pertussis toxin (PT) and filamentous haemagglutinin (FHA), the pertactin is precipitated by ammonium sulphate and the precipitate is dissolved in buffer at pH 6.0 to 8.5, the solution then is passed through hydroxyapatite and ion-exchange chromatograph columns before final ultrafiltration. Cells are extracted with urea and the extract ultrafiltered and diafiltered. The pertactin is precipitated from the extract and the precipitate processed as above. In a variation, the broth is contacted with ammonium sulphate to precipitate pertactin, PT and FHA, the precipitate is dissolved and the PT and FHA selectively removed, before the solution is passed to the chromatograph columns.

    摘要翻译： 从稳定的生物纯的形式中回收Pertactin（以前的69kDa的蛋白质），其发酵培养液中不含可检测的腺苷酸环化酶活性，来自百日咳博德特氏菌以及细胞的发酵。 处理肉汤以选择性地除去百日咳毒素（PT）和丝状血凝素（FHA），通过硫酸铵沉淀百日咳素，将沉淀物溶于pH 6.0-8.5的缓冲液中，然后将溶液通过羟基磷灰石和离子交换 最后超滤前的色谱柱。 用尿素萃取细胞，超滤和渗滤提取物。 从提取物中沉淀出促黄体生成素，并如上所述处理沉淀物。 在一个变化中，将肉汤与硫酸铵接触以沉淀pertactin，PT和FHA，沉淀物溶解，并且在溶液通过色谱柱之前选择性地除去PT和FHA。