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Publication numberCN103691154 A
Publication typeApplication
Application numberCN 201310644051
Publication date2 Apr 2014
Filing date5 Dec 2013
Priority date5 Dec 2013
Also published asCN103691154B
Publication number201310644051.3, CN 103691154 A, CN 103691154A, CN 201310644051, CN-A-103691154, CN103691154 A, CN103691154A, CN201310644051, CN201310644051.3
Inventors朱辉, 陶永祥
Applicant成都雅途生物技术有限公司
Export CitationBiBTeX, EndNote, RefMan
External Links: SIPO, Espacenet
Chromatographic column for protein purification
CN 103691154 A
Abstract
The invention provides a chromatographic column for protein purification, belonging to the field of biological purification instruments. According to the chromatographic column provided by the invention, the column body is provided with scales, so that the operating personnel can conveniently control the filler height and feed amount; and the three silica gel hoses are arranged on the inner wall of the column body, so that a buffer solution, a sample solution, an eluate and other liquids can conveniently flow down along the wall of the column body, thereby avoiding splashing. The chromatographic column has the advantages of simple structure and favorable protein purification effect, and is convenient to operate.
Claims(2)  translated from Chinese
1.一种用于蛋白纯化的层析柱,包括柱头(I)和柱体(7),所述柱头(I)与柱体(7)为螺纹连接,其特征在于,所述柱头(I)的顶部设有供塑料管(2)通过的通孔;所述柱体(7)采用透明的塑料材质,所述柱体(7)上沿柱体轴向设置有均匀的刻度(4);所述塑料管(2)伸入柱体(7)内的底部连接一个四通管,所述四通管的另外三端分别连接一个硅胶软管(6),所述硅胶软管(6)的另一端与柱体(7)的内壁接触;所述柱体(7)的底部为锥形出口(3);所述锥形出口(3)的上部设置筛板(5),所述的筛板(5)的孔径为0.0.45 μ m0 1. A chromatography column for the purification of proteins, including the stigma (I) and the cylinder (7), the stigma (I) and the cylinder (7) for the threaded connection, characterized in that the stigma (I ) provided for the plastic top tube (2) through the through hole; said cylinder (7) using a transparent plastic material, said cylinder (7) along the cylinder, provided with a uniform scale (4) ; bottom (2) extending into the cylinder (7) inside the plastic tube connected to a four-way pipe, the other three of the four ends of the tube are connected to a silicone hose (6), the silicone hose (6 ) the other end of the inner wall of the cylinder (7) of the contact; bottom of said cylinder (7) is tapered outlet (3); the upper part of the conical outlet (3) is provided sieve (5), the the sieve (5) of a pore size of 0.0.45 μ m0
2.根据权利要求1所述的用于蛋白纯化的层析柱,其特征在于,所述柱头(I)和柱体(7)的连接处设有密封圈,所述的密封圈为硅胶材质。 The column used for protein purification according to claim 1, characterized in that the stigma (I) and the cylinder (7) is provided at the connection ring, said ring is silicone material .
Description  translated from Chinese

一种用于蛋白纯化的层析柱 A chromatography column for protein purification

[0001] [0001]

技术领域 FIELD

[0002] 本发明属于生物纯化器械,具体涉及一种用于蛋白纯化的层析柱。 [0002] The present invention belongs to the biological purification device, in particular to a chromatography column used for protein purification.

背景技术[0003]目前,多种化合物的组合物、天然产物及中药的分离纯化,主要是采用柱色谱,使用层析柱,通过柱层析过程来实现。 BACKGROUND OF THE INVENTION [0003] Currently, the composition of a variety of compounds, separation and purification of natural products and medicine, mainly by column chromatography using column by column chromatography procedures to implement. 传统的柱色谱是将混合化合物放置在玻璃柱柱床的顶端,使混合溶剂组成的洗脱剂(流动相)在重力作用下,通过装有吸附性介质材料(固定相)的垂直玻璃柱,分离得到各个较纯的产品(纯品),其原理是:由于混合化合物中各组分对色谱柱中固定相和流动相的亲和力不同,故根据混合化合物的性质选择合适的固定相和流动相,可以达到将其中各组分尽可能分离的目的,分离的产品在柱子出口分别被收集到各个收集瓶中,然后,鉴定各个收集瓶中的收集到的层析液,根据鉴定结果合并和浓缩各相同层析液,得到各个纯品。 Conventional column chromatography is to mix the compound placed on top of a glass column packed bed, the mixed solvent composition of the eluent (mobile phase) under gravity, through the dielectric material containing adsorbent (stationary phase) of a vertical glass column, each isolated pure product (pure), the principle is: Because each component of the mixed compound affinity column stationary and mobile phases are different, so choose the appropriate fixed and mobile phases depending on the nature of the mixed compound , which can achieve the purpose of the separation of the components as much as possible, the separation of the product were collected at the column outlet to the respective collection bottle, and then, the identification of each collecting vial collected liquid chromatography, based on the identification results combined and concentrated isotropic liquid chromatography to give each pure.

[0004] 常见层析柱根据压力不同可分为常压柱、中压柱和高压柱。 [0004] According to common chromatography atmospheric pressure can be divided into columns, medium pressure and high pressure column column. 常压柱在实际中用得最多,常常是自己根据需要采用不同材料的层析柱、装填不同的填料来实现目标物质的分离纯化。 In the actual atmospheric column most frequently used, often their own column using different materials needed, different filler loading to achieve separation and purification of the target substance. 玻璃层析柱又尤其常用,常包括柱体、活塞,有的还包括位于柱体上端的用于固定加液球的倒钩等。 Glass column also particularly popular, often including cylinders, pistons, and some also include the upper cylinder located barbs and other fixed dosing for the ball. 目前常见的玻璃层析柱存在如下不足:一方面,在玻璃层析柱的填料装填和被分离物质样品装填中,常常根据不同填料和不同物质采用不同的填料,样品比例,以利于在达到最佳分离效果的同时节约填料,而填料.样品的比例通常参照其体积比,但是目前常用玻璃柱没有刻度,常常需要操作人员凭经验目测或用其他度量工具进行测量,非常麻烦;另一方面,在层析过程中,常常需要掌握样品在分离介质即填料中的迁移速度,然后根据迁移速度进行流出液的速度调节,以利于在达到最佳分离效果的同时节约层析时间,但是目前的层析柱没有刻度,常常是在不同时间点对某个层析色带的位置用记号笔标记,这样的做法不够准确,同时标记笔所标记的记号可能会因为添加洗脱剂时的泄漏而被冲刷掉,需要再次标记,很不方便。 Existing common glass column following disadvantages: on the one hand, a glass column packed in packing and loading of the material sample are separated, often using different packing according to different fillers and different substances, the proportion of the sample in order to facilitate reaching the most Good separation while saving filler, and filler are usually referring to the proportion of the sample volume ratio, but there is no common scale glass columns, often require the operator to visually or measured empirically, very troublesome to use other measurement tools; on the other hand, In the chromatographic process, it is often necessary to grasp the sample separating medium i.e. filler migration speed, then the speed of the effluent in accordance with the migration speed adjustment, in order to facilitate optimal chromatographic separation while saving time, but the current layer No scale analytical column, often at different time points for a chromatography ribbon position marked with a marker, such an approach is not accurate enough, while marking pen marks of identification may be added as the eluent when the leak is washed away, you need to re-mark, which was very convenient.

[0005] 此外,目前的层析柱在进样时,样品溶液从层析柱顶端的中部直接流进柱体,容易造成柱体内填料飞溅至层析柱的内壁上。 [0005] In addition, the current column during injection, the sample solution flows into the cylinder from the middle of the top of the column directly, easily lead to the body of the column packing to the inner wall of the splash chromatography column. 由于层析用的填料常常是比较昂贵的,飞溅至内壁上的填料没有在层析中得到充分的利用,影响层析效果。 Since chromatography filler is often relatively expensive, splash onto the inner wall of the filler material has not been fully utilized in chromatography, chromatography impact effect.

发明内容 SUMMARY

[0006] 为解决上述问题,本发明提供一种用于蛋白纯化的层析柱,采用如下技术方案: 一种用于蛋白纯化的层析柱,包括柱头和柱体,所述柱头与柱体为螺纹连接,其特征在 [0006] To solve the above problems, the present invention provides a method for protein purification by column chromatography, using the following technical solution: A method for protein purification by column chromatography, including stigma and cylinder, said cylinder stigma and for threaded connection, characterized in

于,所述柱头的顶部设有供塑料管通过的通孔;所述柱体采用透明的塑料材质,所述柱体上沿柱体轴向设置有均匀的刻度;所述塑料管伸入柱体内的底部连接一个四通管,所述四通管的另外三端分别连接一个硅胶软管,所述硅胶软管的另一端与柱体的内壁接触;所述柱体的底部为锥形出口;所述锥形出口的上部设置筛板,所述的筛板的孔径为0.1-0.45 μ m。 At the top of the stud is provided with a through hole through the plastic tube for; the cylinder using transparent plastic material, disposed on the cylinder along the cylinder, a uniform scale; the plastic tube is put into the column at the bottom of the body of a four-way pipe connecting the four-way pipe ends are connected to the other three a silicone hose, silicone hose and the other end of the contact with the inner wall of the cylinder; the cylinder bottom conical outlet ; upper part of the conical outlet is set sieve, sieve apertures for the 0.1-0.45 μ m. [0007] 优选地,所述柱头和柱体的连接处设有密封圈,所述的密封圈为硅胶材质。 [0007] Preferably, the junction of the stigma and the cylinder has a seal, the seal of silicone material.

[0008] 本发明具有如下有益效果: [0008] The present invention has the following advantages:

1.本发明提供的层析柱,其柱体采用透明的塑料材质,并且在柱体上设置均匀的刻度,如此设置,有利于在层析柱的填料装填和样品上样时方便准确地把握填料样品比例,通过观察某个色带在不同时间点所在的刻度位置,从而方便掌握被分离物质在层析柱中的迁移速率,使整个层析过程更方便,提高分离效率和整个工作效率。 Column 1. The present invention provides, its cylinder using transparent plastic material, and is provided on the cylinder in a uniform scale, thus provided, is conducive to easily and accurately grasp when the sample and the sample loaded onto the packing of the column Filling sample ratio, by observing a ribbon at different points in time where the scale positions, thus facilitating control the rate of migration in the substances to be separated in the column, so that the whole chromatography process is more convenient, to improve the overall separation efficiency and work efficiency.

[0009] 2.本发明提供的层析柱,在塑料管伸入柱体内的底部连接一个四通管,所述四通管的另外三端分别连接一个硅胶软管,所述硅胶软管的另一端与柱体的内壁接触,如此设置,有利于缓冲液、样品液、洗脱液从柱体顶部进入时,是沿着柱体的侧壁流下的,不会造成柱体内液体的飞溅,引起填料飞溅在柱体的侧壁上,影响填料高度,从而影响蛋白的纯化效 [0009] 2. The present invention provides a chromatography column, the column at the bottom of the plastic tube is put into the body of a four-way pipe connection, the crossbar of the other three terminals are connected to a silicone hose, a silicone hose and the other end in contact with the inner wall of the cylinder, so provided, a buffer conducive to sample liquid, at the top of the column into the eluate from the column is flowing down along the side wall, without causing splashing of the liquid column body, cause packing splash on the sidewall of the cylinder, the impact of packing height, thus affecting the efficiency of protein purification

果O If O

[0010] 3.本发明提供的层析柱,在锥形出口的上部设置筛板,并限定筛板的孔径为 [0010] 3. The present invention provides a chromatography column, is provided in the upper conical outlet sieve, and the sieve aperture is defined

0.1~0.45 μ m,如此这样便可以达到蛋白纯化时的填料不会随着缓冲液、样品液、洗脱液一起,从锥形出口流失,但又不会影响缓冲液、样品液和洗脱液从锥形出口的流出效果。 0.1 ~ 0.45 μ m, so this will be able to reach a filler protein purification are not together, from the tapered outlet drain with buffer, sample solution, eluent, but will not affect the buffer solution, the sample solution and elution outflow of fluid from the outlet cone effect.

[0011] 4.本发明提供的层析柱,在柱头和柱体的连接处设有密封圈,便于柱头和柱体的密闭,避免缓冲液、样品液、洗脱液从两者的缝隙处流出,造成浪费。 [0011] The present invention provides a chromatography column, is provided at the junction of the stigma and cylinder seals, easy stigma and cylinder closed, to avoid buffer, sample solution, the eluate from the crevice between the two outflow, resulting in waste.

[0012] 5.本发明提供的层析柱,结构简单,操作方便,蛋白的纯化效果好。 [0012] 5. The present invention provides a chromatography column, simple structure, easy operation, good protein purification effect.

附图说明 Brief Description

[0013] 图1为本发明的结构示意图;其中,I表示柱头;2表示塑料管;3表示锥形出口;4表不刻度;5表不筛板;6表不硅胶软管;7表不柱体。 [0013] Figure 1 a schematic view of the structure of the present invention; wherein, I represents stigma; 2 represents a plastic tube; 3 showing the tapered outlet; no scale Table 4; Table 5 no sieve; no silicone hose Table 6; Table 7 does not cylinder.

具体实施方式 DETAILED DESCRIPTION

[0014] 下面结合附图和实施例对本发明作进一步说明。 [0014] accompanying drawings and the following embodiment of the present invention is further illustrated. 除下述实施方式外,本发明还可以有其他实施方式,凡采用等同替换或等效变换形成的技术方案,均落在本发明要求的保护范围内。 In addition to the following embodiment, the present invention is capable of other embodiments, where the use of equivalents or equivalent transformation formed aspect, fall within the scope of the invention claimed.

[0015] 如图1所示,一种用于蛋白纯化的层析柱,包括柱头1和柱体7,所述柱头1与柱体7为螺纹连接,其特征在于,所述柱头1的顶部设有供塑料管2通过的通孔;所述柱体7采用透明的塑料材质,所述柱体7上沿柱体轴向设置有均匀的刻度4 ;所述塑料管2伸入柱体7内的底部连接一个四通管,所述四通管的另外三端分别连接一个硅胶软管6,所述硅胶软管6的另一端与柱体7的内壁接触;所述柱体7的底部为锥形出口3 ;所述锥形出口3的上部设置筛板5,所述的筛板5的孔径为0.1 "0.45 μ m。 [0015] As shown, a method for protein purification column 1, column 1 and 7 comprises a stigma, a stigma with the cylinder 7 is screwed, characterized in that the top of stigma 1 2 is provided for the through hole through the plastic tube; the cylinder 7 using transparent plastic material, the cylinder 7 along the axial direction of the cylinder 4 is provided with a uniform scale; the plastic pipe 2 extends into the cylinder 7 at the bottom of a four-way pipe connections within the four-pipe ends are connected to the other three a silicone hose 6, the other end of the silicone hose and the inner wall of the cylinder 6, 7 contact; the bottom of the column 7 tapered outlet 3; upper portion of said conical outlet 3 is provided sieve 5, wherein the sieve aperture 5 is 0.1 "0.45 μ m.

[0016] 作为优选,在柱头1和柱体7的连接处设有密封圈在图中并没有标注出来,用于密封作用,所述的密封圈为硅胶材质,密封效果更佳。 [0016] Advantageously, the connection is provided at the stigma and column 7 of seal 1 in the figure and not marked out for the seal, the seal is silicone material, the sealing better.

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Classifications
International ClassificationB01D15/22, C07K1/16
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