公开(公告)号：US5728553A

公开(公告)日：1998-03-17

申请号：US378859

申请日：1995-05-25

申请人： Andrew R. Goodey ,  Darrell Sleep ,  Hendrik van Urk ,  Stephen Berezenko ,  John R. Woodrow ,  Richard A. Johnson ,  Patricia C. Wood ,  Steven J. Burton ,  Alan V. Quirk ,  David St. J. Coghlan ,  Mark J. Wilson

发明人： Andrew R. Goodey ,  Darrell Sleep ,  Hendrik van Urk ,  Stephen Berezenko ,  John R. Woodrow ,  Richard A. Johnson ,  Patricia C. Wood ,  Steven J. Burton ,  Alan V. Quirk ,  David St. J. Coghlan ,  Mark J. Wilson

IPC分类号： C12P21/02 ,  A61K9/08 ,  A61K47/42 ,  A61P7/08 ,  A61P17/02 ,  C07K14/76 ,  C07K14/765 ,  C12N1/19 ,  C12N15/14 ,  B01J39/06 ,  B01J41/06

CPC分类号： C07K14/70571 ,  C07K14/765 ,  C07K2319/10

摘要： A process for the preparation of albumin which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, fragments of albumin, polymers or aggregates of albumin, and viruses, and which is relatively non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises passing albumin (preferably expressed and secreted by transformed yeast) through two chromatography purifications, ultrafiltering the product, passing through two further chromatography steps and again ultrafiltering the product.

摘要翻译： 一种制备白蛋白的方法，其具有极低的水平或基本上不含着色剂，金属离子，人类蛋白质，白蛋白片段，白蛋白聚合物或聚集体和病毒，并且相对非糖化，相对较高 游离硫醇和完整的C-末端。 该方法包括使白蛋白（优选由转化酵母表达和分泌）通过两次色谱纯化，超滤产物，通过另外两个色谱步骤，并再次超滤产物。

公开(公告)号：US5667986A

公开(公告)日：1997-09-16

申请号：US270076

申请日：1994-07-01

申请人： Andrew R. Goodey ,  Darrell Sleep ,  Dina Vakeria

发明人： Andrew R. Goodey ,  Darrell Sleep ,  Dina Vakeria

IPC分类号： C07K14/765 ,  C12N9/34 ,  C12N15/81 ,  C12P21/06 ,  C12N1/19 ,  C12N15/11 ,  C12N15/63

CPC分类号： C12N9/2428 ,  C07K14/765 ,  C12N15/81

摘要： A yeast promoter comprising the promoter of the (cytoplasmically located glycerol-3-phosphate dehydrogenase gene (GPD 1) possesses advantageous capacity to regulate the expression of heterologous polypeptides in yeasts transformed therewith. Expression of such polypeptides utilizing the GPD1 promoter can be regulated by the presence(repressed) or absence(derepressed) of high levels of sucrose or glucose in the fermentation medium. Alternatively, a non-repressing carbon source, such as glycerol or ethanol, can be added to the fermentation medium.

摘要翻译： 包含（细胞质定位的甘油-3-磷酸脱氢酶基因（GPD 1））的启动子的酵母启动子具有调节异源多肽在转化的酵母中的表达的有利能力，利用GPD1启动子的这种多肽的表达可以通过 在发酵培养基中存在（抑制）或不存在（去抑制）高水平的蔗糖或葡萄糖。或者，可以向发酵培养基中加入非抑制性碳源，例如甘油或乙醇。

公开(公告)号：US5302697A

公开(公告)日：1994-04-12

申请号：US67243

申请日：1993-05-21

申请人： Andrew R. Goodey ,  Graham P. Belfield ,  Darrell Sleep

发明人： Andrew R. Goodey ,  Graham P. Belfield ,  Darrell Sleep

IPC分类号： C12N15/09 ,  A61K38/00 ,  C07K14/00 ,  C07K14/39 ,  C07K14/76 ,  C07K14/765 ,  C07K19/00 ,  C12N1/19 ,  C12N15/62 ,  C12N15/81 ,  C12P21/02 ,  C12R1/645 ,  C12R1/865 ,  A61K37/00 ,  A61K37/02 ,  C07K7/00 ,  C12P21/06

CPC分类号： C07K14/765 ,  C07K14/39 ,  C12N15/625 ,  C12N15/81 ,  C07K2319/02 ,  C07K2319/036 ,  C07K2319/50 ,  C07K2319/90

摘要： Secretory leader sequences, for use in secreting heterologous polypeptides in yeast, are formed by fusing part of the human serum albumin pre-sequence or part of the Kluyveromyces lactis killer toxin pre-sequence to the Saccharomyces cerevisiae mating factor alpha-1 KEX2 cleavage recognition site. The resulting fusion leader sequences are:(a) H.sub.2 N-Met-Lys-Trp-Val-Ser-Phe-Ile-Ser-Leu-Leu-Phe-Leu-Phe-Ser-Ser-Ala-Tyr-Ser-Arg-Ser-Leu-Asp-Lys-Arg-COOH or(b) H.sub.2 N-Met-Asn-Ile-Phe-Tyr-Ile-Phe-Leu-Phe-Leu-Leu-Ser-Phe-Val-Gln-Gly-Ser-Leu-Asp-Lys-Arg-COOHConservative variations are also encompassed.

摘要翻译： 用于分泌酵母中的异源多肽的分泌型前导序列通过将部分人血清白蛋白前序列或部分乳酸克鲁维酵母杀伤毒素前序序融合到酿酒酵母交配因子α-1 KEX2切割识别位点而形成 。 所得的融合前导序列是：（a）H 2 N-Met-Lys-Trp-Val-Ser-Phe-Ile-Ser-Leu-Leu-Phe-Leu-Phe-Ser-Ser-Ala-Tyr-Ser-Arg- Ser-Leu-Asp-Lys-Arg-COOH或（b）H 2 N-Met-Asn-Ile-Phe-Tyr-Ile-Phe-Leu-Phe-Leu-Leu-Ser-Phe-Val-Gln-Gly-Ser -Leu-Asp-Lys-Arg-COOH也包括保守的变体。

公开(公告)号：US5766883A

公开(公告)日：1998-06-16

申请号：US153799

申请日：1993-11-17

申请人： David J. Ballance ,  Andrew R. Goodey

发明人： David J. Ballance ,  Andrew R. Goodey

IPC分类号： C07K14/765 ,  C12N15/62 ,  C07K19/00 ,  C12N5/10 ,  C12P21/00

CPC分类号： C12N15/62 ,  C07K14/765 ,  C07K2319/02 ,  C07K2319/31 ,  C07K2319/50

摘要： A fusion polypeptide comprising, as at least part of the N-terminal portion thereof, an N-terminal portion of HSA or a variant thereof and, as at least part of the C-terminal portion thereof, another polypeptide except that, when the said N-terminal portion of HSA is the 1-n portion where n is 369 to 419 or a variant thereof then the said polypeptide is one of various specified entities. The HSA-like portion may have additional N-terminal residues, such as secretion leader sequences (signal sequences). The C-terminal portion is preferably the amino terminal fragment of human urokinase-type plasminogen activator. The N-terminal and C-terminal portions may be cleavable to yield the isolated C-terminal portion, with the N-terminal portion having served to facilitate secretion from the host. Such cleavage can be achieved in yeast using a sequence cleavable by the KEX2 protease of S. cerevisiae.

摘要翻译： 一种融合多肽，其至少部分包含HSA的N-末端部分或其变体，并且至少部分其C-末端部分包含另外的多肽，除了当所述 HSA的N-末端部分是n-369〜419的1-n部分或其变体，则所述多肽是各种指定实体之一。 HSA样部分可以具有另外的N-末端残基，例如分泌前导序列（信号序列）。 C-末端部分优选为人尿激酶型纤溶酶原激活物的氨基末端片段。 N-末端和C-末端部分可以是可切割的以产生分离的C-末端部分，其中N末端部分有助于促进从宿主分泌。 可以使用可由酿酒酵母的KEX2蛋白酶切割的序列在酵母中实现这种切割。