公开(公告)号：US07858361B2

公开(公告)日：2010-12-28

申请号：US10587155

申请日：2005-01-26

申请人： Hyung-Kwon Lim ,  Jung Hwan Park ,  Sung-Geun Kim

发明人： Hyung-Kwon Lim ,  Jung Hwan Park ,  Sung-Geun Kim

IPC分类号： C12N1/00

CPC分类号： C12N15/81 ,  C07K14/755

摘要： The present invention relates to a method for preparing LK8 protein, more precisely, a method for mass-production of LK8 protein using Saccharomyces cerevisiae transformed by a gene coding LK8 protein having angiogenesis inhibiting activity. The transformed strain of the present invention and production processes of LK8 protein are expected to contribute greatly to the commercialization of LK8 protein as a novel angiogenesis inhibitor.

摘要翻译： 本发明涉及一种用于制备LK8蛋白的方法，更确切地说，涉及使用由编码具有血管发生抑制活性的LK8蛋白的基因转化的酿酒酵母来批量生产LK8蛋白的方法。 预期本发明的转化菌株和LK8蛋白的生产过程对于作为新型血管发生抑制剂的LK8蛋白质的商业化有很大贡献。

公开(公告)号：US20130045871A1

公开(公告)日：2013-02-21

申请号：US13634359

申请日：2011-03-18

申请人： Matthew DeLisa ,  Hyung-Kwon Lim ,  Amy Karlsson

发明人： Matthew DeLisa ,  Hyung-Kwon Lim ,  Amy Karlsson

IPC分类号： C40B10/00 ,  C40B40/02 ,  C40B30/04

CPC分类号： C12N15/1037 ,  C07K2319/02 ,  C07K2319/033 ,  C12N15/1058 ,  G01N33/6854

摘要： The present invention provides systems, vectors and methods for isolation of enhanced ligand-binding proteins from combinatorial libraries displayed on the inner membrane of a host cell.

摘要翻译： 本发明提供用于从显示在宿主细胞内膜上的组合文库中分离增强的配体结合蛋白的系统，载体和方法。

公开(公告)号：US08298787B2

公开(公告)日：2012-10-30

申请号：US12305768

申请日：2006-06-20

申请人： Hyung-Kwon Lim ,  Sung-Geun Kim

发明人： Hyung-Kwon Lim ,  Sung-Geun Kim

IPC分类号： C12P21/06 ,  C12P21/04

CPC分类号： C12N15/81

摘要： Provided is a method for improving secretion efficiency of a recombinant foreign protein in a yeast expression system. The method includes transforming a yeast host with a recombinant foreign gene construct containing a galactose-inducible promoter, a secretion signal sequence and a gene encoding the foreign protein to construct a transformed yeast strain; and culturing the transformed yeast strain under the condition that the activity of the galactose-inducible promoter is controlled. Improved secretion efficiency of the foreign protein can be achieved by decreasing overexpression-induced insoluble precipitation of the recombinant foreign protein suffered by a conventional galactose-inducible promoter-based yeast expression system, via appropriate control of a level of galactose functioning as an inducer of the galactose-inducible promoter in cells. Due to improved secretion efficiency of the recombinant foreign protein, present invention makes a contribution to improvement in productivity of recombinant foreign proteins in the yeast expression system and reduction in production costs.

摘要翻译： 提供了一种用于提高酵母表达系统中重组外源蛋白的分泌效率的方法。 该方法包括用包含半乳糖诱导型启动子，分泌信号序列和编码外源蛋白的基因的重组外源基因构建体来转化酵母宿主以构建转化的酵母菌株; 并在控制半乳糖诱导型启动子的活性的条件下培养转化的酵母菌株。 外源蛋白的分泌效率的提高可以通过减少由常规的基于半乳糖诱导型启动子的酵母表达系统所承受的重组外源蛋白的过度表达诱导的不溶性沉淀，通过适当控制作为诱导剂的诱导剂的半乳糖的水平来实现 半乳糖诱导型启动子。 由于重组外源蛋白的分泌效率提高，本发明对于提高酵母表达系统中重组外源蛋白质的生产力和生产成本的降低有所贡献。

公开(公告)号：US10078085B2

公开(公告)日：2018-09-18

申请号：US14422912

申请日：2012-08-22

申请人： Hyung-Kwon Lim ,  Sung Geun Kim ,  Young Seoub Park ,  Hyo Jung Nam ,  Dong-Sik Kim ,  Jae Chan Park ,  Yeup Yoon

发明人： Hyung-Kwon Lim ,  Sung Geun Kim ,  Young Seoub Park ,  Hyo Jung Nam ,  Dong-Sik Kim ,  Jae Chan Park ,  Yeup Yoon

IPC分类号： C40B40/10 ,  G01N33/68 ,  C07K16/00 ,  C07K16/18 ,  C07K16/32

CPC分类号： G01N33/6857 ,  C07K16/00 ,  C07K16/18 ,  C07K16/32 ,  C07K2317/21 ,  C07K2317/56 ,  C07K2317/565 ,  C07K2317/569 ,  C07K2317/94 ,  C07K2319/00 ,  C07K2319/10 ,  C40B40/10 ,  G01N2500/00

摘要： There are provided a method named Tat-associated protein engineering (TAPE), of screening a target protein having higher solubility and excellent thermostability, in particular, an immunoglobulin variable domain (VH or VL) derived from human germ cells, by preparing a gene construct where the target protein and an antibiotic-resistant protein are linked to a Tat signal sequence, and then expressing this within E. coli, and human or engineered VH and VL domain antibodies and human or engineered VH and VL domain antibody scaffolds having solubility and excellent thermostability, which are screened by the TAPE method. There are also provided a library including random CDR sequences in the human or engineered VH or VL domain antibody scaffold screened by the TAPE method, and a preparing method thereof. There are also provided a VH or VL domain antibody having binding ability to the target protein screened by using the library, and a pharmaceutical composition including the domain antibody.

公开(公告)号：US20110129872A1

公开(公告)日：2011-06-02

申请号：US12628767

申请日：2009-12-01

申请人： Hyung Kwon LIM ,  Jin-Ho SEO ,  Yong-Cheol PARK

发明人： Hyung Kwon LIM ,  Jin-Ho SEO ,  Yong-Cheol PARK

IPC分类号： C12P21/06

CPC分类号： C12N9/90 ,  C07K2319/02 ,  C12N9/60 ,  C12N15/81

摘要： The present invention relates to a method for mass production of a recombinant protein comprising the step of culturing a yeast transformed with: a recombinant gene construct comprising a yeast promoter, a gene coding a signal sequence and a gene coding a target protein; and also with one or more genes coding folding accessory protein selected from the group consisting of PDI1 (protein disulfide isomerase 1), SEC23 (secretory 23), TRX2 (thioredoxin 2) AHA1 (activator of heat shock protein 90 ATPase), and SCJ1 (S. cerevisiae DnaJ) followed by culturing the transformed yeast.

摘要翻译： 本发明涉及一种大规模生产重组蛋白的方法，包括以下步骤：培养转化酵母的步骤：重组基因构建体，其包含酵母启动子，编码信号序列的基因和编码靶蛋白的基因; 以及选自PDI1（蛋白质二硫键异构酶1），SEC23（分泌型23），TRX2（硫氧还蛋白2））AHA1（热休克蛋白90 ATP酶的活化剂）和SCJ1（ 酿酒酵母DnaJ），然后培养转化的酵母。

公开(公告)号：US20100184136A1

公开(公告)日：2010-07-22

申请号：US12305768

申请日：2006-06-20

申请人： Hyung-Kwon Lim ,  Sung-Geun Kim

发明人： Hyung-Kwon Lim ,  Sung-Geun Kim

IPC分类号： C12P21/00

CPC分类号： C12N15/81

摘要： Provided is a method for improving secretion efficiency of a recombinant foreign protein in a yeast expression system. The method comprises transforming a yeast host with a recombinant foreign gene construct comprising a galactose-inducible promoter, a secretion signal sequence and a gene encoding the foreign protein to construct a transformed yeast strain; and culturing the transformed yeast strain under the condition that the activity of the galactose-inducible promoter is controlled. Improved secretion efficiency of the foreign protein can be achieved by decreasing over-expression-induced insoluble precipitation of the recombinant foreign protein suffered by a conventional galactose-inducible promoter-based yeast expression system, via appropriate control of a level of galactose functioning as an inducer of the galactose-inducible promoter in cells. Due to improved secretion efficiency of the recombinant foreign protein, present invention makes a contribution to improvement in productivity of recombinant foreign proteins in the yeast expression system and reduction in production costs.

摘要翻译： 提供了一种用于提高酵母表达系统中重组外源蛋白的分泌效率的方法。 该方法包括用包含半乳糖诱导型启动子，分泌信号序列和编码外源蛋白的基因的重组外源基因构建体来转化酵母宿主以构建转化的酵母菌株; 并在控制半乳糖诱导型启动子的活性的条件下培养转化的酵母菌株。 通过适当控制用作诱导剂的半乳糖水平，通过减少由常规的基于半乳糖诱导型启动子的酵母表达系统所承受的重组外源蛋白质的过表达诱导的不溶性沉淀，可以实现外源蛋白质的分泌效率的提高。 的半乳糖诱导型启动子。 由于重组外源蛋白的分泌效率提高，本发明对于提高酵母表达系统中重组外源蛋白质的生产力和生产成本的降低有所贡献。

公开(公告)号：US20080187940A1

公开(公告)日：2008-08-07

申请号：US10587155

申请日：2005-01-26

申请人： Hyung-Kwon Lim ,  Jung Hwan Park ,  Sung-Geun Kim

发明人： Hyung-Kwon Lim ,  Jung Hwan Park ,  Sung-Geun Kim

IPC分类号： G01N33/569 ,  C12N15/63 ,  C12P21/02 ,  C12N1/19

CPC分类号： C12N15/81 ,  C07K14/755

摘要： The present invention relates to a method for preparing LK8 protein, more precisely, a method for mass-production of LK8 protein using Saccharomyces cerevisiae transformed by a gene coding LK8 protein having angiogenesis inhibiting activity. The transformed strain of the present invention and production processes of LK8 protein are expected to contribute greatly to the commercialization of LK8 protein as a novel angiogenesis inhibitor.

摘要翻译： 本发明涉及一种用于制备LK8蛋白的方法，更确切地说，涉及使用由编码具有血管发生抑制活性的LK8蛋白的基因转化的酿酒酵母来批量生产LK8蛋白的方法。 预期本发明的转化菌株和LK8蛋白的生产过程对于作为新型血管发生抑制剂的LK8蛋白质的商业化有很大贡献。