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    • 4. 发明申请
    • BIOLOGICAL MOLECULES AND METHODS OF USE
    • 生物分子和使用方法
    • WO2014205491A1
    • 2014-12-31
    • PCT/AU2014/000662
    • 2014-06-25
    • MONASH UNIVERSITYTHE WALTER AND ELIZA HALL INSTITUTE OF MEDICAL RESEARCH
    • NETTER, HansCHEONG, Wan-ShooKINGSTON, NatalieCAMINSCHI, IrinaLAHOUD, Mireille
    • A61K39/44A61K35/76A61P31/20C07K19/00
    • C07K16/2851A61K2039/505A61K2039/5258C07K16/1063C07K2317/76C07K2319/00C07K2319/735
    • A recombinant VLP-antibody (VLP-Ab) comprising a modified hepadnaviral envelope fusion protein comprising an antibody variable domain. The variable domain may be selected from an antibody heavy chain variable domain and a light chain variable domain. In some embodiments, the variable domain (the first variable domain) is bound to a complementary antibody variable domain of a second variable domain protein and forms the antibody antigen binding site specific for a target molecule. One second variable domain protein may be a modified hepadnaviral envelope fusion protein comprising a complementary antibody variable domain. The VLP-Ab may comprises a modified hepadnaviral envelope fusion protein comprising a heterologous antigen. One modified hepadnaviral envelope fusion protein is a hepatitis B envelope protein or a VLP-forming part or variant thereof. A eukaryotic expression vector comprising a recombinant nucleic acid molecule encoding a hepadnavirus envelope-antibody fusion protein comprising an antibody variable domain wherein the variable domain is selected from an antibody heavy chain variable domain and a light chain variable domain. A eukaryotic expression vector comprising a recombinant nucleic acid molecule encoding a second variable domain protein comprising an antibody variable domain wherein the variable domain is selected from an antibody heavy chain variable domain and a light chain variable domain. The expression vector may encode a single variable domain selected from an antibody heavy chain variable domain and a light chain variable domain. The expression vector may encode an Sc Fv. A eukaryotic expression vector operably linked to a polynucleotide encoding a hepadnavirus envelope-antibody fusion protein comprising and an antibody variable domain (EAFP) wherein the antibody variable domain is selected from an antibody heavy chain variable domain and a light chain variable domain, and further capable of expressing a second polynucleotide encoding a second variable domain protein comprising a complementary antibody variable domain. An isolated eukaryotic cell comprising the expression vector. Pharmaceutical and diagnostic compositions are provided and a wide range of therapeutic or prophylactic uses and methods. The manufacture of a medicament for, the treatment or prevention of an infection of a subject by a pathogen, or an inflammatory, immunity or cancer-related condition in a subject. A method for preparing a VLP-Ab, the method comprising transfecting a eukaryotic cell with an expression vector as described, culturing the cell in vitro for a time and under conditions sufficient to permit VLP-Ab formation and optionally secretion, and at least partly purifying the VLP-Ab from the cell culture material.
    • 包含修饰的肝炎病毒包膜融合蛋白的重组VLP-抗体(VLP-Ab),其包含抗体可变结构域。 可变结构域可以选自抗体重链可变结构域和轻链可变结构域。 在一些实施方案中,可变结构域(第一可变结构域)与第二可变结构域蛋白质的互补抗体可变结构域结合,并形成针对靶分子特异性的抗体抗原结合位点。 一秒可变结构域蛋白可以是包含互补抗体可变结构域的修饰的肝炎病毒包膜融合蛋白。 VLP-Ab可以包含包含异源抗原的经修饰的肝炎病毒包膜融合蛋白。 一种修饰的肝炎病毒包膜融合蛋白是乙型肝炎包膜蛋白或其形成VLP的部分或变体。 包含编码包含抗体可变结构域的肝炎病毒包膜 - 抗体融合蛋白的重组核酸分子的真核表达载体,其中所述可变结构域选自抗体重链可变结构域和轻链可变结构域。 包含编码包含抗体可变结构域的第二可变结构域蛋白质的重组核酸分子的真核表达载体,其中所述可变结构域选自抗体重链可变结构域和轻链可变结构域。 表达载体可以编码选自抗体重链可变结构域和轻链可变结构域的单个可变结构域。 表达载体可以编码Sc Fv。 可编码与编码肝炎病毒包膜 - 抗体融合蛋白的多核苷酸可操作地连接的真核表达载体和抗体可变结构域(EAFP),其中抗体可变结构域选自抗体重链可变结构域和轻链可变结构域, 表达编码包含互补抗体可变结构域的第二可变结构域蛋白质的第二多核苷酸。 包含表达载体的分离的真核细胞。 提供药物和诊断组合物以及广泛的治疗或预防用途和方法。 在受试者中制造用于治疗或预防受试者受病原体的感染或炎症,免疫或癌症相关病症的药物。 一种制备VLP-Ab的方法,所述方法包括用如上所述的表达载体转染真核细胞,在体外培养细胞一段时间并在足以允许VLP-Ab形成和任选分泌的条件下,并至少部分纯化 来自细胞培养物质的VLP-Ab。