公开(公告)号：US5093241A

公开(公告)日：1992-03-03

申请号：US250962

申请日：1988-09-23

申请人： Alan D. Bennett ,  Stephen K. Rhind ,  Peter A. Lowe ,  Christopher C. G. Hentschel

发明人： Alan D. Bennett ,  Stephen K. Rhind ,  Peter A. Lowe ,  Christopher C. G. Hentschel

IPC分类号： C12N15/00 ,  A61K39/00 ,  A61K39/385 ,  A61K39/395 ,  C07H1/00 ,  C07H21/04 ,  C07K1/22 ,  C07K14/585 ,  C07K14/695 ,  C07K17/02 ,  C07K19/00 ,  C12N1/20 ,  C12N9/10 ,  C12N15/09 ,  C12P21/02 ,  C12R1/19

CPC分类号： C12N9/1033 ,  A61K39/00 ,  C07K14/585 ,  C07K14/695 ,  C07K2319/00 ,  C07K2319/40 ,  C07K2319/50 ,  C07K2319/61 ,  C07K2319/75

摘要： A process for the production of a fusion protein comprising an active portion of a chloramphenicol acetyltransferase (CAT) protein and a polypeptide. The fusion protein may be purified using CAT substrate affinity chromatography. The eucaryotic polypeptide may be calcitonin or a dervative thereof such as calcitonin-glycine. Other polypeptides described include enzymes such as chymosin, prochymosin and preprochymosin, hormones such as ACTH, insulins, and growth hormones and antigenic polypetides such as foot and mouth disease virus antigenic polypetide. The fusion protein may be cleaved at a site susceptible to selective enzymic or chemical cleavage to produce free polypeptide. The fusion protein may be used as an immunogen.

摘要翻译： 一种生产包含氯霉素乙酰转移酶（CAT）蛋白的活性部分和多肽的融合蛋白的方法。 可以使用CAT底物亲和层析纯化融合蛋白。 真核多肽可以是降钙素或其降糖素，例如降钙素 - 甘氨酸。 所述的其它多肽包括酶，例如凝乳酶，凝乳酶原和前凝乳酶原，激素如ACTH，胰岛素和生长激素以及抗原性多肽，例如口蹄疫病毒抗原性多肽。 融合蛋白可能在易于选择性酶切或化学切割的位点处被切割以产生游离多肽。 融合蛋白可以用作免疫原。

公开(公告)号：US5340926A

公开(公告)日：1994-08-23

申请号：US102697

申请日：1993-08-05

申请人： Peter A. Lowe ,  Fiona A. O. Marston ,  Sarojani Angal ,  Joyce A. Schoemaker

发明人： Peter A. Lowe ,  Fiona A. O. Marston ,  Sarojani Angal ,  Joyce A. Schoemaker

IPC分类号： C07K1/113 ,  C12N9/64 ,  C07K3/12 ,  C07K15/06

CPC分类号： C12N9/6481 ,  C07K1/1136

摘要： In a process for the production of a soluble native protein, such as immunoglobulin or methionine-prochymosin, in which an insoluble form of the protein is produced by a host organism transformed with a vector including a gene coding for the protein, the insoluble form of the protein is reversibly denatured in an alkaline aqueous solution at a pH selected to promote dissociation of a group or groups of the protein involved in maintaining the conformation of the protein, and the protein is subsequently allowed to renature by reducing the pH of the solution below a pH effective to denature the protein to produce the soluble native form of the protein. The pH of the alkaline aqueous is suitably in the range 9.0 to 11.5.

摘要翻译： 在生产可变天然蛋白质例如免疫球蛋白或甲硫氨酸凝乳酶原的方法中，其中蛋白质的不溶形式由宿主生物体转化，宿主生物体用包含编码蛋白质的基因的载体转化， 该蛋白质在选择用于促进参与维持蛋白质构象的一组或多组蛋白质的解离的pH下在碱性水溶液中可逆地变性，并且随后通过降低下述溶液的pH来使蛋白质复性 有效地使蛋白质变性以产生可溶性天然形式的蛋白质的pH。 碱性水溶液的pH适宜为9.0〜11.5。

公开(公告)号：US5332805A

公开(公告)日：1994-07-26

申请号：US677415

申请日：1991-03-29

申请人： Norman H. Carey ,  Michael T. Doel ,  Timothy J. R. Harris ,  Peter A. Lowe

发明人： Norman H. Carey ,  Michael T. Doel ,  Timothy J. R. Harris ,  Peter A. Lowe

IPC分类号： C12N9/64 ,  C07K3/12 ,  C07K15/06

CPC分类号： C12N9/6481

摘要： The production of recombinant chymosin is disclosed in which an insoluble form of chymosin precursor is produced by a bacterial host cell transformed by a vector including a coding sequence for said precursor. Solubilization of said insoluble form of chymosin precursor is accomplished using urea at a concentration of at least 7M or guanidine hydrochloride at a concentration of at least 6M prior to cleaving said precursor to form chymosin. Said solubilization preferably additionally involves the denaturation of said precursor in an alkaline aqueous solution, e.g., at a pH between a 9 and 11.5

摘要翻译： 公开了重组凝乳酶的生产，其中由包含所述前体的编码序列的载体转化的细菌宿主细胞产生不溶性形式的凝乳酶前体。 所述不溶形式的凝乳酶前体的溶解是在切割所述前体以形成凝乳酶之前，使用至少7M浓度的尿素或至少6M浓度的盐酸胍。 所述溶解优选另外包括在碱性水溶液中，例如在9和11之间的pH下变性所述前体。