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    • 5. 发明申请
    • MAJOR OUTER MEMBRANE PROTEIN CD OF MORAXELLA
    • MORAXELLA主要外膜蛋白CD
    • WO1996012733A1
    • 1996-05-02
    • PCT/CA1995000604
    • 1995-10-23
    • CONNAUGHT LABORATORIES LIMITEDYANG, Yan-PingHARKNESS, Robin, E.MYERS, Lisa, E.McGUINNESS, UrsulaKLEIN, Michel, H.
    • CONNAUGHT LABORATORIES LIMITED
    • C07K14/22
    • C07K14/212A61K39/00
    • An isolated and purified non-denatured outer membrane protein CD which is that of or corresponds to that isolatable from a Moraxella strain, particularly M. catarrhalis, is isolated from a bacterial strain by fractionating a cell lysate formed by disrupting a cell mass of the bacterial strain by centrifugation to provide a pellet and a discard supernatant containing a large proportion of soluble bacterial proteins. The pellet is selectively extracted to remove the remaining soluble proteins, the membrane proteins other than CD and other contaminants such as lipopolysaccharide and phospholipids. The remaining CD-containing pellet is dispersed and solubilized and then fractionated by centrifugation to remove the remaining cell debris. The CD protein is useful in diagnostic applications and immunogenic compositions, particularly for in vivo administration to a host to confer protection against disease caused by a bacterial pathogen that produces CD protein or produces a protein capable of inducing antibodies in a host specifically reactive with CD protein.
    • 分离纯化的非变性外膜蛋白CD,其是与莫拉氏菌属菌株特别是卡他莫拉菌可分离的非变性外膜蛋白质CD,通过分离通过破坏细菌的细胞块而形成的细胞裂解物从细菌菌株中分离 通过离心以提供含有大部分可溶性细菌蛋白质的沉淀物和丢弃上清液。 选择性提取沉淀物以除去剩余的可溶性蛋白质,除CD以外的膜蛋白质和其它污染物如脂多糖和磷脂。 剩余的含有CD的颗粒被分散和溶解,然后通过离心分离分离以除去剩余的细胞碎片。 CD蛋白可用于诊断应用和免疫原性组合物,特别是用于体内施用宿主,以产生由产生CD蛋白的细菌病原体引起的疾病的保护,或产生能够在与CD蛋白特异性反应的宿主中诱导抗体的蛋白质 。
    • 10. 发明申请
    • OUTER MEMBRANE PROTEIN P1 AND PEPTIDES OF HAEMOPHILUS INFLUENZAE TYPE B
    • 外膜蛋白P1和HAEMOPHILUS流感病毒B型胶原蛋白
    • WO1991006652A1
    • 1991-05-16
    • PCT/CA1990000374
    • 1990-10-31
    • CONNAUGHT LABORATORIES LIMITEDWASHINGTON UNIVERSITY, ST. LOUISMUNSON, Robert, S., Jr.GRASS, SusanCHONG, PeleYANG, Yan-PingFAHIM, RaafatSTA, Dwo, Yan, CharlesMcVERRY, PatrickKLEIN, Michel
    • CONNAUGHT LABORATORIES LIMITEDWASHINGTON UNIVERSITY, ST. LOUIS
    • C12N15/31
    • C07K14/285A61K39/00
    • 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.
    • 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。