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    • 9. 发明申请
    • PRODUCTION OF ANTIBODIES OR (FUNCTIONALIZED) FRAGMENTS THEREOF DERIVED FROM HEAVY CHAIN IMMUNOGLOBULINS OF CAMELIDAE
    • 生产抗原或其功能性片段,其衍生自CAMELIDAE重链免疫球蛋白
    • WO1994025591A1
    • 1994-11-10
    • PCT/EP1994001442
    • 1994-04-28
    • UNILEVER PLCUNILEVER N.V.HAMERS, RaymondHAMERS-CASTERMAN, CécileMUYLDERMANS, Serge, Victor, M.FRENKEN, Leon, Gerardus, J.VERRIPS, Cornelis, Theodorus
    • UNILEVER PLCUNILEVER N.V.
    • C12N15/13
    • C07K16/00C07K2319/00C07K2319/02
    • A process is provided for the production of an antibody or a fragment or functionalized fragment thereof using a transformed lower eukaryotic host containing an expressible DNA sequence encoding the antibody or (functionalized) fragment thereof, wherein the antibody or (functionalized) fragment thereof is derived from a heavy chain immunoglobulin of Camelidae and is devoid of light chains, and wherein the lower eukaryotic host is a mould, preferably belonging to the genera Aspergillus or Trichoderma, or a yeast, preferably belonging to the yeast genera Saccharomyces, Kluyveromyces, Hansenula, or Pichia. The heavy chain fragment can contain at least the whole variable domain. A complementary determining region (CDR) different from the CDR belonging to the natural antibody ex Camelidae can be grafted on the framework of the variable domain of the heavy chain immunoglobulin. The catalytic antibodies can be raised in Camelidae against transition state molecules. The functionalized antibody or fragment thereof can comprise a fusion protein of both a heavy chain immunoglobulin from Camelidae or a fragment thereof and another polypeptide, e.g., an enzyme, preferably an oxido-reductase. Also provided are new products obtainable by a process as described, and compositions containing a product produced by a process as described, which composition may contain a new product as provided.
    • 提供了使用含有编码抗体或其(功能化的)片段的可表达DNA序列的转化的低等真核宿主制备抗体或其片段或功能化片段的方法,其中抗体或其(官能化)片段衍生自 骆驼科的重链免疫球蛋白,没有轻链,其中低等真核宿主是模具,优选属于曲霉属或木霉属,或酵母,优选属于酵母属酵母属,酵母属,汉逊酵母属或毕赤酵母属 。 重链片段至少可以包含整个可变结构域。 可以将不同于属于骆驼科目天然抗体的CDR的互补决定区(CDR)移植到重链免疫球蛋白的可变区的框架上。 催化抗体可以在Camelidae中针对过渡态分子产生。 功能化抗体或其片段可以包含来自骆驼科的重链免疫球蛋白或其片段和另一多肽(例如酶,优选氧化还原酶)的融合蛋白。 还提供了可通过所述方法获得的新产品,以及含有通过所述方法生产的产品的组合物,该组合物可含有所提供的新产品。
    • 10. 发明申请
    • IMPROVEMENTS IN THE EXPRESSION OF NEWLY INTRODUCED GENES IN YEAST CELLS
    • 在YEAST细胞中新引入基因表达的改进
    • WO1984004538A1
    • 1984-11-22
    • PCT/EP1984000153
    • 1984-05-21
    • UNILEVER N.V.EDENS, LuppoRUSSELL, Stephen, WilliamVISSER, ChristiaanVERRIPS, Cornelis, Theodorus
    • UNILEVER N.V.
    • C12N15/00
    • C12N9/6481C07K14/43C07K2319/00C07K2319/02C12N15/625C12N15/81C12N15/815
    • The invention relates to an improved expression system for newly introduced genes in yeast and comprises a yeast regulon, and preferably a transcription terminator, derived from one of the GAPDH genes of S. cerevisiae. The about 850 nucleotides long GAPDH regulon described proved to be almost ten times as effective as smaller regulons described previously. Said regulon and/or terminator can be introduced into yeasts either as part of plasmide molecules or by incorporating into the yeast genome. Vectors containing the expression system preferably comprises an autonomously replicating sequence derived from K. lactis (KARS) or an origin of replication originating from the S. cerevisiae 2 micron yeast plasmid. After transformation of yeasts, in particular of the genera Kluyveromyces and Saccharomyces, with said vectors, the yeasts transformed can produce foreign proteins more effectively. In the case of thaumatin the presence of a signal polypeptide (the pre-part) appears to be essential for expression in yeast. DNA sequences encoding other signal polypeptides are also described. The use of codons preferred by yeast in both structural genes, preferably those encoding thaumatin-like proteins and chymosin-like proteins, and DNA sequences encoding signal polypeptides is also described.
    • 本发明涉及一种用于酵母中新引入的基因的改进的表达系统,其包含衍生自酿酒酵母的GAPDH基因之一的酵母调节子,并且优选是转录终止子。 所描述的大约850个核苷酸长的GAPDH调节子证明比以前描述的较小的调节子的几十倍有效。 所述调节子和/或终止子可以作为浆细胞分子的一部分引入酵母中,或通过掺入酵母基因组中。 含有表达系统的载体优选包含衍生自乳酸克鲁维酵母(KARS)的自主复制序列或源自酿酒酵母2微米酵母质粒的复制起点。 在酵母转化后,尤其是克鲁维酵母属和酵母属,用所述载体转化后,转化的酵母可以更有效地产生外源蛋白。 在奇异果蛋白的情况下,信号多肽(前体部分)的存在似乎对酵母中表达是必需的。 还描述了编码其他信号多肽的DNA序列。 还描述了在两个结构基因中优选使用密码子的密码子,优选编码类似蛋白质的蛋白质和凝乳酶样蛋白质的那些,以及编码信号多肽的DNA序列。