公开(公告)号：US07541164B2

公开(公告)日：2009-06-02

申请号：US10742564

申请日：2003-12-18

Applicant: Bernhard M. Schilling ,  Linda Matlock ,  Stephen G. Zegarelli ,  William V. Burnett ,  Christoph E. Joosten ,  Jonathan D. Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

Inventor： Bernhard M. Schilling ,  Linda Matlock ,  Stephen G. Zegarelli ,  William V. Burnett ,  Christoph E. Joosten ,  Jonathan D. Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

IPC: C12N5/06 ,  A61K39/395 ,  A61K38/00

CPC classification number: C12P21/005 ,  C07K14/70521 ,  C12N5/0018 ,  C12N2500/34 ,  C12N2500/74 ,  C12N2501/33 ,  C12N2501/90 ,  C12N2501/905 ,  C12N2501/91 ,  C12N2510/02

Abstract: The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, preferably, but not limited to, fed-batch cell cultures. In one aspect, the methods comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. Throughout their duration, the culturing processes of the invention involving two or more downward shifts in temperature sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product, as determined, e.g., by sialic acid content of the produced protein. In another aspect, the methods comprise the delayed addition of polyanionic compound during the culturing period. The delayed addition of polyanionic compound sustains a high viability of the cultured cells, and can extend the growth phase, delay the onset of the death phase, and arrest the death phase.

Abstract translation: 本发明描述了通过动物细胞或哺乳动物细胞培养物生产蛋白质，特别是糖蛋白的方法和方法，优选但不限于补料分批细胞培养物。 在一个方面，所述方法包括在培养期间进行的至少两个温度漂移，其中在培养期结束时温度比初始细胞培养时的温度更低。 在整个持续时间内，涉及两个或更多个温度向下移位的本发明的培养方法维持培养细胞的高活力，并且可以产生蛋白质产物的最终滴定度和蛋白质产物的高质量，如所确定的，例如 ，通过产生蛋白质的唾液酸含量。 另一方面，所述方法包括在培养期间延迟加入聚阴离子化合物。 聚阴离子化合物的延迟添加维持培养细胞的高活力，并且可以延长生长期，延缓死亡阶段的发作并停止死亡期。

公开(公告)号：US07332303B2

公开(公告)日：2008-02-19

申请号：US10740645

申请日：2003-12-18

Applicant: Bernhard M. Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen G. Zegarelli ,  Christoph E. Joosten ,  Jonathan D. Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

Inventor： Bernhard M. Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen G. Zegarelli ,  Christoph E. Joosten ,  Jonathan D. Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

IPC: C12N15/09 ,  C12N15/06 ,  C12N15/56 ,  A61K38/00

CPC classification number: C07K14/70521 ,  C12N2500/34 ,  C12P21/005

Abstract: The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, illustratively, but not limited to, fed-batch cell cultures. The methods comprise feeding the cells with D-galactose, preferably with feed medium containing D-galactose, preferably daily, to sustain a sialylation effective level of D-galactose in the culture for its duration, thus increasing sialylation of the produced proteins. The methods can also comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. The cell culture processes of the invention involving two or more temperature shifts sustain a high cell viability, and can allow for an extended protein production phase. The methods can also comprise the delayed addition of polyanionic compound at a time after innoculation. Supplementation of the cultures with D-galactose, preferably in a feed medium, to sustain galactose at sialylation effective levels in the cultures until the end of a culture run reverses a decline in sialylation that accompanies culture scale up, and is advantageous for large scale culturing processes.

Abstract translation: 本发明描述了通过动物细胞或哺乳动物细胞培养生产蛋白质，特别是糖蛋白的方法和方法，例如但不限于补料分批细胞培养物。 所述方法包括用D-半乳糖喂养细胞，优选用含有D-半乳糖的饲料培养基，优选每天喂养，以维持培养基中唾液酸化有效水平的D-半乳糖的持续时间，从而增加产生的蛋白质的唾液酸化。 所述方法还可以包括在培养期间进行的至少两个温度漂移，其中在培养期结束时温度比初始细胞培养时温度更低。 涉及两个或多个温度变化的本发明的细胞培养方法维持高的细胞活力，并且可以允许延长的蛋白质生产阶段。 所述方法还可以包括在接种后延迟加入聚阴离子化合物。 用D-半乳糖，优选在饲料培养基中补充培养物，以维持培养物中唾液酸化有效水平的半乳糖，直至培养结束时反转伴随培养物扩大的唾液酸化的下降，并且有利于大规模培养 过程。

公开(公告)号：US20050084933A1

公开(公告)日：2005-04-21

申请号：US10740645

申请日：2003-12-18

Applicant: Bernhard Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen Zegarelli ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

Inventor： Bernhard Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen Zegarelli ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

IPC: C07K14/705 ,  C12P21/00 ,  C12P21/02 ,  C12N5/06

CPC classification number: C07K14/70521 ,  C12N2500/34 ,  C12P21/005

Abstract: The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, illustratively, but not limited to, fed-batch cell cultures. The methods comprise feeding the cells with D-galactose, preferably with feed medium containing D-galactose, preferably daily, to sustain a sialylation effective level of D-galactose in the culture for its duration, thus increasing sialylation of the produced proteins. The methods can also comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. The cell culture processes of the invention involving two or more temperature shifts sustain a high cell viability, and can allow for an extended protein production phase. The methods can also comprise the delayed addition of polyanionic compound at a time after innoculation. Supplementation of the cultures with D-galactose, preferably in a feed medium, to sustain galactose at sialylation effective levels in the cultures until the end of a culture run reverses a decline in sialylation that accompanies culture scale up, and is advantageous for large scale culturing processes.

Abstract translation: 本发明描述了通过动物细胞或哺乳动物细胞培养物生产蛋白质，特别是糖蛋白的方法和方法，例如但不限于补料分批细胞培养物。 所述方法包括用D-半乳糖喂养细胞，优选用含有D-半乳糖的饲料培养基，优选每天喂养，以维持培养基中唾液酸化有效水平的D-半乳糖的持续时间，从而增加产生的蛋白质的唾液酸化。 所述方法还可以包括在培养期间进行的至少两个温度漂移，其中在培养期结束时温度比初始细胞培养时温度更低。 涉及两个或多个温度变化的本发明的细胞培养方法维持高的细胞活力，并且可以允许延长的蛋白质生产阶段。 所述方法还可以包括在接种后延迟加入聚阴离子化合物。 用D-半乳糖，优选在饲料培养基中补充培养物，以维持培养物中唾液酸化有效水平的半乳糖，直至培养结束时反转伴随培养物扩大的唾液酸化的下降，并且有利于大规模培养 过程。

公开(公告)号：US06593178B1

公开(公告)日：2003-07-15

申请号：US08866968

申请日：1997-06-02

Applicant: Steven S. Lee

Inventor： Steven S. Lee

IPC: H01L218238

CPC classification number: H01L21/8249

Abstract: The invention concerns a BI-CMOS process, in which Field-Effect Transistors (FETs) and Bipolar Junction Transistors (BJTs) are manufactured on a common substrate. In several processing steps, FET structures are formed simultaneously with BJT structures. For example, in one step, polysilicon gate electrodes for the FETs and polysilicon emitters for the BJTs are formed simultaneously. In another aspect of the invention, a polysilicon layer is used to reduce channeling which would otherwise occur during an implant step.

公开(公告)号：US5734119A

公开(公告)日：1998-03-31

申请号：US769400

申请日：1996-12-19

Applicant: Gordon Scott France ,  Steven S. Lee

Inventor： Gordon Scott France ,  Steven S. Lee

IPC: G10H1/00 ,  G10H1/06 ,  G10H7/00

CPC classification number: G10H1/0066 ,  G10H1/0058 ,  G10H1/0075 ,  G10H2240/305

Abstract: An Internet high fidelity audio transmission and compression protocol including a system for representing synthesized music in a relatively small file as compared to digital recording. The protocol includes a method for streaming the transmission of a music data file from a Server-Composer computer such that the music can begin being played back as soon as the file begins to arrive at a Client-Player computer. The system includes a graduated resolution improvement feature which allows the music to be recreated exactly as originally composed as the necessary wavetable data is downloading in the background and the music continues to play in the foreground.

Abstract translation: 一种互联网高保真音频传输和压缩协议，包括与数字记录相比在相对小的文件中表示合成音乐的系统。 该协议包括用于从服务器 - 作曲家计算机流式传输音乐数据文件的方法，使得一旦文件开始到达客户端 - 播放器计算机就可开始播放音乐。 该系统包括分级分辨率改进功能，其允许按照最初的组合重新创建音乐，因为必要的波表数据正在背景中下载并且音乐在前景中继续播放。

公开(公告)号：US5543361A

公开(公告)日：1996-08-06

申请号：US351843

申请日：1994-12-08

Applicant: Steven S. Lee ,  Kenneth P. Fuchs ,  Gayle W. Miller

Inventor： Steven S. Lee ,  Kenneth P. Fuchs ,  Gayle W. Miller

IPC: H01L21/3205 ,  H01L21/28 ,  H01L21/336 ,  H01L21/768 ,  H01L23/52 ,  H01L29/78 ,  H01L21/44

CPC classification number: H01L21/76895 ,  H01L2924/0002 ,  Y10S148/015 ,  Y10S148/019 ,  Y10S148/147

Abstract: A process for forming a titanium silicide local interconnect between electrodes separated by a dielectric insulator on an integrated circuit. A first layer of titanium is formed on the insulator, and a layer of silicon is formed on the titanium. The silicon layer is masked and etched to form a silicon strip connecting the electrodes, and an overlying second layer of titanium is formed over the silicon strip. The titanium and silicon are heated to form nonsilicidized titanium over a strip of titanium silicide, and the nonsilicidized titanium is removed.

公开(公告)号：US5516718A

公开(公告)日：1996-05-14

申请号：US331235

申请日：1994-10-25

Applicant: Steven S. Lee

Inventor： Steven S. Lee

IPC: H01L21/8249

CPC classification number: H01L21/8249 ,  Y10S148/009 ,  Y10S148/124

Abstract: The invention concerns a BI-CMOS process, in which Field-Effect Transistors (FETs) and Bipolar Junction Transistors (BJTs) are manufactured on a common substrate. In several processing steps, FET structures are formed simultaneously with BJT structures. For example, in one step, polysilicon gate electrodes for the FETs and polysilicon emitters for the BJTs are formed simultaneously. In another aspect of the invention, a polysilicon layer is used to reduce channeling which would otherwise occur during an implant step.

Abstract translation: 本发明涉及一种BI-CMOS工艺，其中在公共衬底上制造场效应晶体管（FET）和双极结晶体管（BJT）。 在几个处理步骤中，与BJT结构同时形成FET结构。 例如，在一个步骤中，同时形成用于BJT的用于FET的多晶硅栅电极和多晶硅发射极。 在本发明的另一方面，多晶硅层用于减少否则在植入步骤期间会发生的沟道化。

公开(公告)号：US5248350A

公开(公告)日：1993-09-28

申请号：US622107

申请日：1990-11-30

Applicant: Steven S. Lee

Inventor： Steven S. Lee

IPC: H01L21/76 ,  H01L21/316 ,  H01L21/32 ,  H01L21/762

CPC classification number: H01L21/32 ,  H01L21/76221

Abstract: A process for forming field oxide regions between active regions in a semiconductor substrate. Pad oxide, polysilicon and first silicon nitride layers are successively formed over substrate active regions. The first nitride layer, polysilicon layer, pad oxide layer and a portion of the substrate are then selectively etched to define field oxide regions with substantially vertical sidewalls. A second silicon nitride is provided on the substantially vertical sidewalls, and field oxide is grown in the field oxide regions. The first silicon nitride, polysilicon and pad oxide layers are then removed. The presence of the polysilicon layer prevents the formation of a sharp corner between the field oxide and active regions if an overetch occurs during the removal of the pad oxide layer.

公开(公告)号：US5200686A

公开(公告)日：1993-04-06

申请号：US774435

申请日：1991-10-10

Applicant: Steven S. Lee

Inventor： Steven S. Lee

IPC: H02J7/00

CPC classification number: H02J7/0091 ,  H02J7/0006

Abstract: Battery type is determined by measuring effective resistance of a thermistor/resistor network (205, 206, and 207) and one or more of a plurality of current sources (303-305) is enabled to provide the appropriate charging current. Measurement of necessary charging parameters and provision of appropriate charging current are accomplished through an interface to the battery pack undergoing charge that comprises only three connections (313-315).

Abstract translation: 电池类型通过测量热敏电阻/电阻网络（205,206和207）的有效电阻来确定，并且多个电流源（303-305）中的一个或多个能够提供适当的充电电流。 必要的充电参数的测量和适当的充电电流的提供是通过一个接口连接电池组，该电池组正在进行电荷，仅包含三个连接（313-315）。

公开(公告)号：US4855258A

公开(公告)日：1989-08-08

申请号：US111377

申请日：1987-10-22

Applicant: Derryl D. J. Allman ,  Steven S. Lee

Inventor： Derryl D. J. Allman ,  Steven S. Lee

IPC: H01L21/302 ,  H01L21/306 ,  H01L21/318 ,  H01L21/32

CPC classification number: H01L21/02046 ,  H01L21/3185 ,  H01L21/32 ,  Y10S148/081 ,  Y10S148/114 ,  Y10S148/118 ,  Y10S438/906

Abstract: A process for forming a thin sealing layer of silicon nitride directly upon a silicon substrate to minimize bird's beak encroachment. The process employs in situ fabrication whereby the native oxide is removed from the silicon substrate by etching the hydrogen or hydrogen chloride and followed in direct succession, and in the absence of exposure to an oxidizing environment, with the deposition of a silicon nitride layer by LPCVD. Bird's beak encroachment is incrementally reduced by the absence of the native oxide layer as a path for oxygen species movement during the field oxide growth.

Abstract translation: 一种用于在硅衬底上直接形成氮化硅的薄密封层的过程，以最小化鸟的喙侵入。 该方法采用原位制备，其中通过蚀刻氢或氯化氢并直接连续地从硅衬底去除天然氧化物，并且在没有暴露于氧化环境的情况下，通过LPCVD沉积氮化硅层 。 在野外氧化物生长过程中，由于不存在氧化物层作为氧物质运动的路径，鸟的喙侵入逐渐减少。