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    Synthesis of protein with an identification peptide, and hybrid polypeptide incorporating same
    2.
    发明授权
    Synthesis of protein with an identification peptide, and hybrid polypeptide incorporating same 失效
    用鉴定肽合成蛋白质，以及掺入其的杂交多肽

    公开(公告)号：US4782137A

    公开(公告)日：1988-11-01

    申请号：US76811

    申请日：1987-07-23

    申请人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    发明人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    IPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  A61K37/02 ,  C12P21/00

    CPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  C07K2319/00 ,  C07K2319/40 ,  C07K2319/50

    摘要： A hybrid polypeptide composed of an identification peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the identification peptide and the desired functional protein. The identification peptide consists of a highly antigenic N-terminal portion and a C-terminal linking portion that connects the identification peptide to the N-terminal of the functional protein. The linking portion of the identification peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purfied by affinity chromatography techniques by use of an immobilized ligand specific to the antigenic portion of the identification peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

    摘要翻译： 通过重组DNA技术产生由鉴定肽和所需功能性蛋白质组成的杂合多肽。 构建DNA表达载体，其包括编码鉴定肽和所需功能性蛋白质的DNA片段。 识别肽由高度抗原的N末端部分和连接识别肽与功能性蛋白质的N末端的C-末端连接部分组成。 识别肽的连接部分可通过使用序列特异性蛋白水解酶或化学蛋白水解剂在与功能蛋白相邻的特定氨基酸残基处可切割。 由克隆载体转化的宿主细胞表达的杂合多肽从其中除去，并通过使用对鉴定肽的抗原部分特异性的固定化配体的亲和层析技术进行纯化。 然后用合适的蛋白酶或化学试剂从分离的杂交多肽切割蛋白质，从而释放高度纯化，高活性状态的成熟功能性蛋白质。

    Fatty acid carriers for synthetic peptides
    3.
    发明授权
    Fatty acid carriers for synthetic peptides 失效
    合成肽的脂肪酸载体

    公开(公告)号：US5019383A

    公开(公告)日：1991-05-28

    申请号：US405102

    申请日：1989-09-08

    申请人： Thomas P. Hopp

    发明人： Thomas P. Hopp

    IPC分类号： A61K39/385 ,  A61K39/39 ,  C07K14/02

    CPC分类号： C07K14/005 ,  A61K39/385 ,  A61K39/39 ,  A61K2039/6031 ,  A61K2039/6093 ,  C12N2730/10122

    摘要： A synthetic vaccine is contemplated comprising a peptide residue coupled to one or more alkyl or alkenyl groups of at least 12 carbon atoms or other lipophilic substance wherein said peptide residue contains a sequence of 6 amino acids corresponding to the sequence of such amino acids in a protein antigen or allergen where the greatest local average hydrophilicity of the antigen or allergen is found.

    摘要翻译： 考虑包含与一个或多个至少12个碳原子的烷基或链烯基偶联的肽残基或其它亲脂性物质的合成疫苗，其中所述肽残基含有对应于蛋白质中该氨基酸序列的6个氨基酸的序列 抗原或变应原，其中发现抗原或变应原的最大局部平均亲水性。

    Gene encoding biologically active human interleukin 1
    6.
    发明授权
    Gene encoding biologically active human interleukin 1 失效
    基因编码生物活性人白细胞介素1

    公开(公告)号：US5122459A

    公开(公告)日：1992-06-16

    申请号：US443399

    申请日：1989-11-30

    申请人： Paul J. Conlon, III ,  David j. Cosman ,  Kenneth H. Grabstein ,  Thomas P. Hopp ,  Shirley R. Kronheim ,  Alf D. Larsen ,  Carl J. March ,  Virginia L. Price ,  Douglas P. Cerretti

    发明人： Paul J. Conlon, III ,  David j. Cosman ,  Kenneth H. Grabstein ,  Thomas P. Hopp ,  Shirley R. Kronheim ,  Alf D. Larsen ,  Carl J. March ,  Virginia L. Price ,  Douglas P. Cerretti

    IPC分类号： C07K14/545 ,  C12N15/25 ,  C12N15/81

    CPC分类号： C07K14/545 ,  C12N15/81 ,  Y10S435/942

    摘要： Double-stranded cDNA is prepared from polyadenylated RNA extracted from activated human peripheral blood adherent mononuclear cells. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Transformed hosts are identified and grouped into pools. Plasmid DNA prepared from these pools is hybridized with a labeled, synthetic oligonucleotide probe corresponding to a portion of the amino acid sequence of the interleukin 1 protein. Pools of host cells that provide a positive signal to the probe are identified, plated out and then employed in direct bacterial colony hybridization with the same probe, thereby to isolate the particular positive colony. Plasmid DNA is prepared from this colony and characterized by restriction enzyme mapping and sequencing by chain-termination method. The coding region for the IL-1 gene is inserted into a shuttle vector for amplification of the vector followed by expression of functional IL-1.

    摘要翻译： 双链cDNA由从活化的人外周血粘附单核细胞提取的聚腺苷酸化RNA制备。 将cDNA插入质粒载体中，然后将重组质粒用于转化合适的宿主。 已识别转化的主机并将其分组到池中。 从这些池制备的质粒DNA与对应于白介素1蛋白的氨基酸序列的一部分的标记的合成的寡核苷酸探针杂交。 确定提供探针的正信号的宿主细胞池，电镀，然后用于与相同探针的直接细菌菌落杂交，从而分离出特定的阳性菌落。 从该菌落制备质粒DNA，并通过限制酶作图和通过链终止法进行测序。 将用于IL-1基因的编码区插入穿梭载体中用于扩增载体，随后表达功能性IL-1。

    Synthesis of protein with an identification peptide
    8.
    发明授权
    Synthesis of protein with an identification peptide 失效
    用鉴定肽合成蛋白质

    公开(公告)号：US4703004A

    公开(公告)日：1987-10-27

    申请号：US573825

    申请日：1984-01-24

    申请人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    发明人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    IPC分类号： G01N33/531 ,  A61K35/74 ,  A61K36/06 ,  A61K38/00 ,  A61K39/00 ,  C07C15/04 ,  C07H21/04 ,  C07K14/005 ,  C07K14/195 ,  C07K14/55 ,  C07K16/00 ,  C07K16/44 ,  C07K19/00 ,  C12N5/00 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/62 ,  C12P21/00 ,  C12P21/02 ,  C12P21/08 ,  C12R1/19 ,  C12R1/865 ,  C12R1/91 ,  G01N33/577 ,  C12N1/00

    CPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  C07K2319/00 ,  C07K2319/40 ,  C07K2319/50 ,  Y10S930/28

    摘要： A hybrid polypeptide composed of an identification peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the identification peptide and the desired functional protein. The identification peptide consists of a highly antigenic N-terminal portion and a C-terminal linking portion that connects the identification peptide to the N-terminal of the functional protein. The linking portion of the identification peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purified by affinity chromatography techniques by use of an immobilized ligand specific to the antigenic portion of the identification peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

    摘要翻译： 通过重组DNA技术产生由鉴定肽和所需功能性蛋白质组成的杂合多肽。 构建DNA表达载体，其包括编码鉴定肽和所需功能性蛋白质的DNA片段。 识别肽由高度抗原的N末端部分和连接识别肽与功能性蛋白质的N末端的C-末端连接部分组成。 识别肽的连接部分可通过使用序列特异性蛋白水解酶或化学蛋白水解剂在与功能蛋白相邻的特定氨基酸残基处可切割。 由克隆载体转化的宿主细胞表达的杂合多肽从其中除去，并通过亲和层析技术通过使用对鉴定肽的抗原部分特异性的固定化配体进行纯化。 然后用合适的蛋白酶或化学试剂从分离的杂交多肽切割蛋白质，从而释放高度纯化，高活性状态的成熟功能性蛋白质。