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1. 发明授权

US08058410B2 Affinity ligands 失效
标题翻译：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
申请人： Alois Jungbauer , Rainer Hahn , Waltraud Kaar , Michael Seifert , Bernhard Auer , Clemens Achmüller , Philipp Wechner
发明人： Alois Jungbauer , Rainer Hahn , Waltraud Kaar , Michael Seifert , Bernhard Auer , Clemens Achmüller , Philipp Wechner
IPC分类号： A23J1/00 , C07K1/00 , C07K14/00 , C07K16/00 , C07K17/00 , C07K5/00 , C07K7/00 , A61K38/04
CPC分类号： C12P21/02 , C07K1/22 , C07K17/06 , C12N9/506 , C12N15/62
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。

2. 发明授权

US07829319B2 Glutamic acid-modified classical swine fever virus autoproteases Npro 有权
标题翻译：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
申请人： Florian Werther , Clemens Achmüller , Philipp Wechner , Bernhard Auer , Silvio Podmirseg
发明人： Florian Werther , Clemens Achmüller , Philipp Wechner , Bernhard Auer , Silvio Podmirseg
IPC分类号： C12N9/50 , C12N15/70 , C12N15/74 , C12P21/06 , C12N15/57 , C12N15/62
CPC分类号： C12P21/02 , C07K1/22 , C07K17/06 , C12N9/506 , C12N15/62
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。

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