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    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS
    2.
    发明申请
    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS 审中-公开
    多种基因表达，包括SORF构建体和方法与聚丙烯酸，蛋白质和蛋白质和蛋白质

    公开(公告)号：WO2007014162A3

    公开(公告)日：2009-05-07

    申请号：PCT/US2006028691

    申请日：2006-07-21

    Applicant: ABBOTT LAB ,  CARSON GERALD R ,  GION WENDY ,  SALFELD JOCHEN G ,  GU JIJIE ,  REGIER DEAN A ,  KUNES YUNE

    Inventor： CARSON GERALD R ,  GION WENDY ,  SALFELD JOCHEN G ,  GU JIJIE ,  REGIER DEAN A ,  KUNES YUNE

    IPC: C12N15/63 ,  C12P21/06 ,  C07H21/02 ,  C07H21/04 ,  C07K16/18 ,  C12N1/18 ,  C12N5/07 ,  C12N5/071 ,  C12N15/00 ,  C12P21/04 ,  C12P21/08

    CPC classification number: C12N15/67 ,  C07K16/00 ,  C07K2319/50 ,  C07K2319/92 ,  C12N15/1055 ,  C12P21/02 ,  C12P21/06

    Abstract: Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and/or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

    Abstract translation: 公开了使用在重组宿主细胞内处理的主要翻译产物来表达蛋白质的有用构建体和方法。 描述了包含单个开放阅读框（sORF）的构建体用于蛋白质表达，包括多个多肽的表达。 初级翻译产品（前蛋白质或多蛋白质）含有多肽，例如内含肽或刺猬蛋白家族自身处理结构域或其变体，其插入到感兴趣的多个蛋白质亚基之间。 初级产物还可以含有裂解序列，如其他蛋白水解切割或蛋白酶识别位点，或含有信号肽酶识别序列的信号肽，分离多个蛋白质亚基中的至少两个。 可以操纵插入的自动处理多肽或切割位点的序列，以提高分离的多个蛋白质亚基的表达效率。 还公开了进行蛋白质例如免疫球蛋白的有效表达，分泌和/或多聚体装配的独立方面。 当多蛋白含有免疫球蛋白重链和轻链片段或能够进行抗原识别的片段时，在一个实施方案中，可选择的化学计量比为每个重链段的轻链片段的至少两个拷贝，结果是适当地折叠和组装 制备功能性抗体。 描述了包含免疫球蛋白轻链的修饰的信号肽。

    4.
    发明专利
    未知

    公开(公告)号：BRPI0613784A2

    公开(公告)日：2011-02-01

    申请号：BRPI0613784

    申请日：2006-07-21

    Applicant: ABBOTT LAB

    Inventor： CARSON GERALD R ,  GION WENDY ,  SALFELD JOCHEN G ,  GU JIJIE ,  REGIER DEAN A ,  KUNES YUNE

    IPC: C12N15/63 ,  C12P21/08 ,  C07H21/04 ,  C07K16/18 ,  C12N1/18 ,  C12N5/07 ,  C12N5/071

    Abstract: Disclosed is an expression vector for generating one or more recombinant protein products comprising a promoter regulatory element operably linked to a single open reading frame (sORF) insert; said sORF insert comprising: (a) a first nucleic acid sequence encoding a first polypeptide, (b) a first intervening nucleic acid sequence encoding a first protein cleavage site, and (c) a second nucleic acid sequence encoding a second polypeptide; wherein said intervening nucleic acid sequence encoding said first protein cleavage site is operably positioned between said first nucleic acid sequence and said second nucleic acid sequence; and wherein said expression vector is capable of expressing a sORF polypeptide cleavable at said first protein cleavage site; wherein said first protein cleavage site comprises a self-processing cleavage site comprising a hedgehog segment or modified hedgehog segment, wherein the hedgehog segment or modified hedgehog segment permits cleavage of said sORF polypeptide but not ligation of said first polypeptide and said second polypeptide.

    EXPRESION DE GEN MULTIPLE QUE INCLUYE CONSTRUCCIONES Y METODOS SORF (CUADRO DE LECTURA ABIERTA SIMPLE) CON POLIPROTEINAS, PRO-PROTEINAS Y PROTEOLISIS.
    5.
    发明专利
    EXPRESION DE GEN MULTIPLE QUE INCLUYE CONSTRUCCIONES Y METODOS SORF (CUADRO DE LECTURA ABIERTA SIMPLE) CON POLIPROTEINAS, PRO-PROTEINAS Y PROTEOLISIS. 未知

    公开(公告)号：MX2008000985A

    公开(公告)日：2008-04-07

    申请号：MX2008000985

    申请日：2006-07-21

    Applicant: ABBOTT LAB

    Inventor： CARSON GERALD R ,  GION WENDY ,  SALFELD JOCHEN G ,  GU JIJIE ,  REGIER DEAN A ,  KUNES YUNE

    IPC: C12N15/63 ,  C12P21/08 ,  C07H21/04 ,  C07K16/18 ,  C12N1/18 ,  C12N5/07 ,  C12N5/071

    Abstract: Se describen construcciones y metodos utiles para la expresion de proteinas utilizando productos de traduccion primaria que son procesados dentro de una celula huesped recombinante. Las construcciones que comprenden una sola estructura de lectura abierta (sORF) se describen para la expresion de proteinas incluyendo expresion de polipeptidos multiples. Un producto de traduccion primaria (una proteina o poliproteina) contiene polipeptidos tales como inteinas o dominios de auto-procesamiento de la familia de erizos, o variantes de los mismos, insertadas en la estructura entre multiples subunidades de proteina de interes. El producto primario tambien puede contener secuencias de disociacion, tales como otros sitios de reconocimiento de proteasa o disociacion proteolitica, o peptidos de senal que contienen secuencias de reconocimiento para peptidasas de senal, que separa al menos dos de las multiples subunidades de proteina. Las secuencias de los polipeptidos de auto-procesamiento o sitios de disociacion insertados, pueden manipularse para mejorar la eficiencia de expresion de las subunidades de proteina multiple separadas. Tambien se describen aspectos independientes para llevar a cabo una expresion, secrecion, y/o ensamble de proteinas multimerico eficiente, tal como inmunoglobulinas. Cuando la poliproteina contiene segmentos de cadena pesada y ligera de inmunoglobulina o fragmentos con la capacidad de reconocimiento de antigenos, en una modalidad una proporcion estequiometrica seleccionable es de al menos dos copias de un segmento de cadena ligera por segmento de cadena pesada, obteniendo como resultado que se lleve a cabo la produccion del anticuerpo funcional doblado y ensamblado. Se describen peptidos de senal modificados, incluyendo los que proceden de cadenas ligeras de inmunoglobulina.

    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS
    6.
    发明专利
    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS 未知

    公开(公告)号：CA2615983A1

    公开(公告)日：2007-02-01

    申请号：CA2615983

    申请日：2006-07-21

    Applicant: ABBOTT LAB

    Inventor： GU JIJIE ,  SALFELD JOCHEN G ,  REGIER DEAN A ,  KUNES YUNE ,  GION WENDY ,  CARSON GERALD R

    IPC: C12N15/63 ,  C12P21/08 ,  C07H21/04 ,  C07K16/18 ,  C12N1/18 ,  C12N5/07 ,  C12N5/071

    Abstract: Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and/or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS
    7.
    发明专利
    MULTIPLE GENE EXPRESSION INCLUDING SORF CONSTRUCTS AND METHODS WITH POLYPROTEINS, PRO-PROTEINS, AND PROTEOLYSIS 未知

    公开(公告)号：SG164383A1

    公开(公告)日：2010-09-29

    申请号：SG2010053353

    申请日：2006-07-21

    Applicant: ABBOTT LAB

    Inventor： CARSON GERALD R ,  GION WENDY ,  SALFELD JOCHEN G ,  GU JIJIE ,  REGIER DEAN A ,  KUNES YUNE

    IPC: C12N15/63 ,  C07H21/04 ,  C07K16/18 ,  C12N1/18 ,  C12N5/07 ,  C12N5/071 ,  C12P21/08

    Abstract: Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto- processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and/or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made, Modified signal peptides, including such from immunoglobulin light chains, are described.

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