Glutamic acid-modified classical swine fever virus autoproteases Npro
    1.
    发明授权
    Glutamic acid-modified classical swine fever virus autoproteases Npro 有权
    谷氨酸改性古典猪瘟病毒自体蛋白酶Npro

    公开(公告)号:US07829319B2

    公开(公告)日:2010-11-09

    申请号:US11919346

    申请日:2006-04-25

    摘要: The invention relates to a process for the recombinant production of a heterologous polypeptide of interest by cultivating a bacterial host cell transformed with an expression vector comprising a nucleic acid molecule encoding a fusion polypeptide wherein (a) the amino-proximal fusion partner is an autoprotease Npro comprising the replacement(s) by glutamic acid of one or more cysteines at positions corresponding to the positions 112, 134, and 138 of the autoprotease Npro of classical swine fever virus and (b) the carboxyl-proximal fusion partner is an heterologous polypeptide of interest fused to the autoprotease Npro so that it is capable of being cleaved from the fusion polypeptide by autoprotease Npro autoproteolytic activity, said process comprising (i) cultivating the transformed host cell under conditions permitting the expression of the fusion polypeptide and the formation of corresponding cytoplasmic inclusion bodies, (ii) isolating the inclusion bodies from the host cell, (iii) solubilizing the isolated inclusion bodies, (iv) inducing autoproteolytic cleavage of the heterologous polypeptide of interest from the fusion polypeptide, and (v) isolating the cleaved heterologous polypeptide of interest.

    摘要翻译: 本发明涉及通过培养用包含编码融合多肽的核酸分子的表达载体转化的细菌宿主细胞重组产生感兴趣的异源多肽的方法,其中(a)所述近端融合配偶体是自身蛋白酶Npro 包括在对应于经典猪瘟病毒的自身蛋白酶Npro的位置112,134和138的位置处的谷氨酸替换一个或多个半胱氨酸,和(b)羧基近端融合伴侣是异源多肽 所述方法包括(i)在允许融合多肽表达和形成相应细胞质的条件下培养转化的宿主细胞,所述方法包括:(i)在转化的宿主细胞中培养转化的宿主细胞,其中所述融合多肽通过自身蛋白酶Npro自身蛋白水解活性被切割, 包涵体,(ii)从宿主细胞中分离包涵体,(ii i)增溶分离的包涵体,(iv)从融合多肽诱导感兴趣的异源多肽的自体蛋白水解切割,和(v)分离所述切割的异源多肽。

    Organic Compounds
    2.
    发明申请
    Organic Compounds 有权
    有机化合物

    公开(公告)号:US20100062490A1

    公开(公告)日:2010-03-11

    申请号:US11919346

    申请日:2006-04-25

    IPC分类号: C12P21/04 C12N9/50

    摘要: The invention relates to a process for the recombinant production of a heterologous polypeptide of interest, comprising, (i) cultivation of a bacterial host cell which is transformed with an expression vector which comprises a nucleic acid molecule which codes for a fusion polypeptide, the fusion polypeptide comprising a derivative of an autoprotease Npro of Pestivirus, wherein at least one cysteine residue of the naturally occuring autoprotease Npro of Pestivirus is replaced by another amino acid residue, and a second polypeptide which is connected to the first polypeptide at the C-terminus of the first polypeptide in a manner such, that the second polypeptide is capable of being cleaved from the fusion polypeptide by the autoproteolytic activity of the first polypeptide, said second polypeptide being a heterologous polypeptide, wherein cultivation occurs under conditions which cause expression of the fusion polypeptide and formation of corresponding cytoplasmic inclusion bodies, (ii) isolation of the inclusion bodies from the host cell, (iii) solubilization of the isolated inclusion bodies, (iv) induction of autoproteolytic cleavage of the heterologous polypeptide of interest from the fusion polypeptide, and (v) isolation of the cleaved heterologous polypeptide of interest.

    摘要翻译: 本发明涉及重组产生感兴趣的异源多肽的方法,其包括(i)培养用表达载体转化的细菌宿主细胞,所述表达载体包含编码融合多肽的核酸分子,所述融合 多肽,其包含蚜虫病毒的自身蛋白酶Npro的衍生物,其中所述天然存在的自发蛋白酶Npro的至少一个半胱氨酸残基被另一个氨基酸残基替代,并且第二个多肽与第一个多肽在C末端连接 第一多肽以这样的方式使得第二多肽能够通过第一多肽的自身蛋白水解活性从融合多肽切割,所述第二多肽是异源多肽,其中培养在导致融合多肽表达的条件下发生 和相应的细胞质包涵体的形成,(ii) 从宿主细胞分离包涵体,(iii)分离的包涵体的溶解,(iv)从融合多肽诱导感兴趣的异源多肽的自体蛋白水解切割,和(v)分离感兴趣的异源多肽 。

    Affinity ligands
    3.
    发明授权
    Affinity ligands 失效
    亲和配体

    公开(公告)号:US08058410B2

    公开(公告)日:2011-11-15

    申请号:US11919258

    申请日:2006-04-25

    摘要: Disclosed is an affinity matrix comprising a solid phase and an affinity ligand comprising peptide bonds coupled to this solid phase, wherein the affinity ligand comprising peptide bond is selected from the following group of ligands: a) peptides comprising the formula X1X2X3X4, wherein X1 to X4 are amino acid residues and at least two of X1 to X4 is W, Y or F; b) peptides comprising the formula X5X6X7X8, wherein X5 to X8 are amino acid residues, at least one of X5 to X8 is W, and at least one of X5 to X8 is E or D; and c) poly-amino acids consisting of an amino acid monomer of the group consisting of R, K, E and D and an amino acid monomer of the group consisting of Y, F and W, preferably poly-KY, poly-KF, poly-KW, poly-RY, poly-RF, poly-RW, poly-EY, poly-DY, poly-EF, poly-EW, poly-DF and poly-DW, with the proviso that the peptides according to a) and b) have a maximum length of 35 amino acid residues and that the poly-amino acids according to c) have a minimum length of 20 amino acid residues.

    摘要翻译: 公开了包含固相和包含与该固相偶联的肽键的亲和配体的亲和基质,其中包含肽键的亲和配体选自以下配体组:a)包含式X1X2X3X4的肽,其中X1至X4 是氨基酸残基,X1至X4中的至少两个为W,Y或F; b)包含式X5X6X7X8的肽,其中X5至X8为氨基酸残基,X5至X8中的至少一个为W,X5至X8中的至少一个为E或D; 和c)由R,K,E和D组成的组的氨基酸单体和由Y,F和W组成的组的氨基酸单体组成的聚氨基酸,优选聚KY,聚-KF, poly-KW,poly-RY,poly-RF,poly-RW,poly-EY,poly-DY,poly-EF,poly-EW,poly-DF和poly-DW,条件是根据a) 和b)具有35个氨基酸残基的最大长度,并且根据c)的多氨基酸具有20个氨基酸残基的最小长度。

    Affinity Ligands
    4.
    发明申请
    Affinity Ligands 失效
    亲和力配体

    公开(公告)号:US20090306343A1

    公开(公告)日:2009-12-10

    申请号:US11919258

    申请日:2006-04-25

    IPC分类号: C07K7/06 C07K1/22

    摘要: Disclosed is an affinity matrix comprising a solid phase and an affinity ligand comprising peptide bonds coupled to this solid phase, wherein the affinity ligand comprising peptide bond is selected from the following group of ligands: a) peptides comprising the formula X1X2X3X4, wherein X1 to X4 are amino acid residues and at least two of X1 to X4 is W, Y or F; b) peptides comprising the formula X5X6X7X8, wherein X5 to X8 are amino acid residues, at least one of X5 to X8 is W, and at least one of X5 to X8 is E or D; and c) poly-amino acids consisting of an amino acid monomer of the group consisting of R, K, E and D and an amino acid monomer of the group consisting of Y, F and W, preferably poly-KY, poly-KF, poly-KW, poly-RY, poly-RF, poly-RW, poly-EY, poly-DY, poly-EF, poly-EW, poly-DF and poly-DW, with the proviso that the peptides according to a) and b) have a maximum length of 35 amino acid residues and that the poly-amino acids according to c) have a minimum length of 20 amino acid residues.

    摘要翻译: 公开了包含固相和包含与该固相偶联的肽键的亲和配体的亲和基质,其中包含肽键的亲和配体选自以下配体组:a)包含式X1X2X3X4的肽,其中X1至X4 是氨基酸残基,X1至X4中的至少两个为W,Y或F; b)包含式X5X6X7X8的肽,其中X5至X8为氨基酸残基,X5至X8中的至少一个为W,X5至X8中的至少一个为E或D; 和c)由R,K,E和D组成的组的氨基酸单体和由Y,F和W组成的组的氨基酸单体组成的聚氨基酸,优选聚KY,聚-KF, poly-KW,poly-RY,poly-RF,poly-RW,poly-EY,poly-DY,poly-EF,poly-EW,poly-DF和poly-DW,条件是根据a) 和b)具有35个氨基酸残基的最大长度,并且根据c)的多氨基酸具有20个氨基酸残基的最小长度。

    Antibody molecules and nucleic acids
    5.
    发明授权
    Antibody molecules and nucleic acids 失效
    抗体分子和核酸

    公开(公告)号:US07722869B2

    公开(公告)日:2010-05-25

    申请号:US12150076

    申请日:2008-04-24

    摘要: An scFv peptide comprising a VH domain and a VL domain linked by an amino acid spacer is disclosed. The VH domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 64. The VL domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 66. The scFv peptide also comprises the substitution or deletion of an amino acid in the VH domain at the position corresponding to C28.

    摘要翻译: 公开了包含由VH结构域和通过氨基酸间隔物连接的VL结构域的scFv肽。 VH结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 VL结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 scFv肽还包括在对应于C28的位置上的VH结构域中的氨基酸的取代或缺失。

    NOVEL ANTIBODY MOLECULES AND NUCLEIC ACIDS
    6.
    发明申请
    NOVEL ANTIBODY MOLECULES AND NUCLEIC ACIDS 审中-公开
    新型抗体分子和核酸

    公开(公告)号:US20100166758A1

    公开(公告)日:2010-07-01

    申请号:US12719049

    申请日:2010-03-08

    摘要: An scFv peptide comprising a VH domain and a VL domain linked by an amino acid spacer is disclosed. The VH domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 64. The VL domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 66. The scFv peptide also comprises the substitution or deletion of an amino acid in the VH domain at the position corresponding to C28.

    摘要翻译: 公开了包含由VH结构域和通过氨基酸间隔物连接的VL结构域的scFv肽。 VH结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 VL结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 scFv肽还包括在对应于C28的位置上的VH结构域中的氨基酸的取代或缺失。

    NOVEL ANTIBODY MOLECULES AND NUCLEIC ACIDS BINDING TO FUNGAL STRESS PROTEIN HSP90
    7.
    发明申请
    NOVEL ANTIBODY MOLECULES AND NUCLEIC ACIDS BINDING TO FUNGAL STRESS PROTEIN HSP90 审中-公开
    新型抗体分子和核酸结合真菌应激蛋白HSP90

    公开(公告)号:US20100113355A1

    公开(公告)日:2010-05-06

    申请号:US12597229

    申请日:2006-04-24

    摘要: An scFv peptide comprising a VH domain and a VL domain linked by an amino acid spacer is disclosed. The VH domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 64. The VL domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 66. The scFv peptide comprises an additional feature selected from (a) to (e) and combinations thereof. (a) A substitution or deletion of an amino acid in the VH domain at a position corresponding to that selected from the group consisting of: I29, H68, N85, C97 and combinations thereof. (b) A substitution or deletion of an amino acid in the VL domain at a position corresponding to that selected from the group consisting of: V2, V3, F10, F14, A39, N76 and combinations thereof. (c) The amino acid spacer comprises the sequence (GGGGS)n wherein n is between 4 and 6. (d) The VH domain further comprises an N-terminal pelB signal sequence comprising the sequence of SEQ ID NO. 68 or a sequence having at least 80% sequence identity thereto. (e) The VL domain is located at the N-terminal end of the VH domain. (f) The substitution of the cysteine residue in the VH domain corresponding to position 28 with a serine residue.

    摘要翻译: 公开了包含由VH结构域和通过氨基酸间隔物连接的VL结构域的scFv肽。 VH结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 VL结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 scFv肽包含选自(a)至(e)的附加特征及其组合。 (a)在与选自:I29,H68,N85,C97及其组合的对应位置的VH结构域中的氨基酸的取代或缺失。 (b)在对应于选自V2,V3,F10,F14,A39,N76及其组合的对应位置的VL结构域中的氨基酸的取代或缺失。 (c)氨基酸间隔物包含其中n在4和6之间的序列(GGGGS)n。(d)VH结构域还包含N末端pelB信号序列,其包含SEQ ID NO: 68或与其具有至少80%序列同一性的序列。 (e)VL结构域位于VH结构域的N末端。 (f)用丝氨酸残基取代对应于位置28的VH结构域中的半胱氨酸残基。

    Novel antibody molecules and nucleic acids
    8.
    发明申请
    Novel antibody molecules and nucleic acids 失效
    新型抗体分子和核酸

    公开(公告)号:US20090136484A1

    公开(公告)日:2009-05-28

    申请号:US12150076

    申请日:2008-04-24

    摘要: An scFv peptide comprising a VH domain and a VL domain linked by an amino acid spacer is disclosed. The VH domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 64. The VL domain comprises a sequence with at least 80% sequence identity to the sequence of SEQ ID NO. 66. The scFv peptide also comprises the substitution or deletion of an amino acid in the VH domain at the position corresponding to C28.

    摘要翻译: 公开了包含由VH结构域和通过氨基酸间隔物连接的VL结构域的scFv肽。 VH结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 VL结构域包含与SEQ ID NO:1的序列具有至少80%的序列同一性的序列。 scFv肽还包括在对应于C28的位置上的VH结构域中的氨基酸的取代或缺失。