Method for purification of a protein complex and identification of its components
    7.
    发明授权
    Method for purification of a protein complex and identification of its components 失效
    蛋白质复合物的纯化方法及其成分的鉴定

    公开(公告)号:US07825227B2

    公开(公告)日:2010-11-02

    申请号:US10984958

    申请日:2004-11-09

    IPC分类号: A23J1/00 C12P21/04

    CPC分类号: C07K5/1021 C07K1/22 C07K5/101

    摘要: The present invention provides a method for purifying a protein complex and the components comprising the complex from a cell, a cell or tissue lysate or a whole organism by employing a combined set of affinity tags of high affinity, specificity and ease of elution. The method involves using a protein or a peptide modified to contain one or more affinity tags separated by one or more specific protease cleavage sites to isolate any interacting proteins or fragments thereof. Specifically exemplified is a method employing a modified bait containing AviTag or GST and a removal tag of His6 hexapeptide positioned proximal to the bait such that any excess bait can be efficiently removed from the purified complex yielding enriched interacting proteins prior to subjecting the complex for further identification of individual components. The identification of the proteins or fragments thereof contained in the protein complex will provide new targets for the identification of new pharmaceuticals and diagnostic tools.

    摘要翻译: 本发明提供一种通过使用高亲和力,特异性和容易洗脱的组合的一组亲和标签来纯化蛋白质复合物和包含来自细胞,细胞或组织裂解物或整个生物体的复合物的组分的方法。 所述方法包括使用经修饰的蛋白质或肽含有由一个或多个特异性蛋白酶切割位点分离的一个或多个亲和标签,以分离任何相互作用的蛋白质或其片段。 具体举例说明了使用含有AviTag或GST的修饰诱饵和位于诱饵附近的His6六肽的去除标签的方法,使得任何过量的诱饵可以在将复合物进一步鉴定之前从纯化的复合物中有效地除去产生富集的相互作用蛋白质 的单个组件。 包含在蛋白质复合物中的蛋白质或其片段的鉴定将为鉴定新的药物和诊断工具提供新的目标。

    Multi-well array with adjustable plenum
    8.
    发明授权
    Multi-well array with adjustable plenum 失效
    具有可调节气室的多孔阵列

    公开(公告)号:US06436350B1

    公开(公告)日:2002-08-20

    申请号:US09499068

    申请日:2000-02-04

    IPC分类号: B01L100

    摘要: An apparatus for multiple simultaneous synthesis (110), having a number of vials (28), each vial (28) having an upper end (31), a lower end (30), and a barrel (29), the upper end (31) opening into a mouth (32). It also includes a lower frame assembly (180) having a number of holes (101, 103, 105) for receiving the vials (28), and an upper frame assembly (180), which includes inner side-walls (195) and a ceiling portion (190). A gas-tight seal (197) surrounds a plenum (85), the plenum (85) being formed between the ceiling portion (190) of the upper frame assembly (180) and the lower frame assembly (181). The plenum (85) is adjustable in volume by the positioning of the upper frame assembly (180) in variable relation to the lower frame assembly (181), while maintaining the gas-tight seal (197). The volume of the plenum (85) is adjustable to a minimum whereby the mouths (32) of the vials (28) are sealed by the ceiling (190) of the gas-tight seal (197), which is optionally an upper gasket (90). Also disclosed is a plenum enclosure (182) for multiple simultaneous synthesis used in conjunction with vials (28), which includes the upper frame assembly (180) and lower frame assembly (181) and the gastight seal (197), as well as a gasket vial holder (185) to be used in the plenum enclosure (182).

    摘要翻译: 一种用于多个同时合成(110)的装置,具有多个小瓶(28),每个小瓶(28)具有上端(31),下端(30)和筒(29),所述上端 31)开口(32)。 它还包括具有用于接纳小瓶(28)的多个孔(101,103,105)的下框架组件(180),以及包括内侧壁(195)和 顶棚部分(190)。 气密封(197)围绕增压室(85),所述增压室(85)形成在上框架组件(180)的顶部(190)和下框架组件(181)之间。 在保持气密密封(197)的同时,通过上框架组件(180)与下框架组件(181)的可变关系的定位,气室(85)可以体积调节。 增压室(85)的容积可调节至最小,由此小瓶(28)的口部(32)由气密密封件(197)的顶部(190)密封,气密密封件(197)的顶部可选地是上垫圈 90)。 还公开了一种用于与包括上框架组件(180)和下框架组件(181)和气密密封件(197))的小瓶(28)结合使用的多个同时合成的增压室(182),以及一 密封小瓶架(185),用于增压室外壳(182)。

    Compartmentalized multi-well container
    9.
    发明授权
    Compartmentalized multi-well container 失效
    隔间多孔容器

    公开(公告)号:US5916526A

    公开(公告)日:1999-06-29

    申请号:US945870

    申请日:1997-11-03

    申请人: Paul B. Robbins

    发明人: Paul B. Robbins

    IPC分类号: B01L3/00 B01L3/14

    摘要: A multi-well container (10) is provided in which a greater number of wells may exist than heretofore possible while still maintaining a standard multi-well plate tube array format and footprint. The multi-well container (10) is comprised of a rectangular array of tubes (12) in standard tube format held together by an integrally fashioned plate portion (16). The tubes (12) are subdivided by partitions or septa (14) which extend the height of the tubes (12). Each septum (14) may constitute a single wall or may be comprised of any number of fins (32) which are integral with internal tube surfaces (28). In a symmetrical design with four such fins (32), the fins (32) radiate at angular intervals of 90 degrees from a common central axis (34) with which all four fins (32) are integral. Thus, the septa (14) serve to compartmentalize each tube (12) into symmetrical quadrants of four smaller wells or sub-tubes (30). The preferred embodiment is directed toward usage in conjunction with PCR thermal cyclers, but the multi-well container (10) and the elements as are embodied therein are generally applicable to any laboratory procedure where multiple samples must be treated, evaluated, or stored.

    摘要翻译: PCT No.PCT / US96 / 12985 Sec。 371日期:1997年11月3日 102(e)日期1997年11月3日PCT PCT 1996年8月9日PCT公布。 出版物WO97 / 06890 日期1997年2月27日提供了一种多孔容器(10),其中可能存在比以前更多的孔,同时仍然保持标准的多孔板管阵列格式和占地面积。 多孔容器(10)由标准管形式的由整体形成的板部分(16)保持在一起的矩形管阵列(12)组成。 管(12)通过延伸管(12)的高度的隔板(14)细分。 每个隔膜(14)可以构成单个壁,或者可以由与内部管表面(28)成一体的任何数量的翅片(32)构成。 在具有四个这样的翅片(32)的对称设计中,翅片(32)以与所有四个翼片(32)为一体的公共中心轴线(34)成90度的角度间隔辐射。 因此,隔片(14)用于将每个管(12)分隔成四个较小孔或子管(30)的对称象限。 优选实施方案涉及与PCR热循环仪结合使用,但是多孔容器(10)和其中体现的元件通常可应用于必须处理,评估或储存多个样品的任何实验室程序。

    Locking cap for replaceable needle assembly
    10.
    发明授权
    Locking cap for replaceable needle assembly 有权
    可更换针组件的锁盖

    公开(公告)号:US06364859B1

    公开(公告)日:2002-04-02

    申请号:US09586246

    申请日:2000-05-31

    IPC分类号: A61M5315

    CPC分类号: B01L3/0275

    摘要: A syringe with replaceable needle assembly (10), having a syringe barrel (18) with a proximal end (14), a distal end (16), and an outer surface (87). The syringe barrel (18) surrounds an internal bore (20). A plunger assembly (60) is inserted into the syringe barrel internal bore (20). A needle assembly (80) has a sealing portion (72), which is configured to be removably insertable into the internal bore (20). The sealing portion (72) forms a fluid-tight seal with the internal bore (20). The needle assembly (80) further includes a releasable locking mechanism (79), which includes a collar (76) having a locking flange (86). Also disclosed is a replaceable needle assembly (80), which releasably locks onto the outer surface (87) of the proximal end (14) of a syringe barrel (18). The replaceable needle assembly (80) includes a collar (76), having a locking flange (86), which engages the outer surface (87) of the syringe barrel (18), and releasably locks the replaceable needle assembly (80) onto the syringe barrel (18).

    摘要翻译: 一种具有可更换针组件(10)的注射器,其具有具有近端(14),远端(16)和外表面(87)的注射器筒(18)。 注射器筒(18)围绕内孔(20)。 柱塞组件(60)插入到注射器筒内孔(20)中。 针组件(80)具有密封部分(72),其被构造成可移除地插入内孔(20)中。 密封部分(72)与内孔(20)形成流体密封。 针组件(80)还包括可释放的锁定机构(79),其包括具有锁定凸缘(86)的凸缘(76)。 还公开了一种可更换的针组件(80),其可释放地锁定在注射器筒(18)的近端(14)的外表面(87)上。 可替换针组件(80)包括具有与注射器筒(18)的外表面(87)接合的锁定凸缘(86)的套环(76),并且可释放地将可更换针组件(80) 注射器筒(18)。