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Chromatography column for the purification of proteins

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Publication number
CN103691154B
CN103691154B CN 201310644051 CN201310644051A CN103691154B CN 103691154 B CN103691154 B CN 103691154B CN 201310644051 CN201310644051 CN 201310644051 CN 201310644051 A CN201310644051 A CN 201310644051A CN 103691154 B CN103691154 B CN 103691154B
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CN 201310644051
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CN103691154A (en )
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朱辉
陶永祥
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成都雅途生物技术有限公司
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Abstract

本发明提供一种用于蛋白纯化的层析柱,属于生物纯化器械领域。 The present invention provides a chromatography column for purifying proteins, belonging to the field of biological purification device. 本发明提供的层析柱,柱体上设置刻度,方便操作人员对填料高度和进样量的掌握;与柱体内壁的三个硅胶软管的设置,便于缓冲液、样品液、洗脱液等液体从柱体的四壁流下,避免引起飞溅。 The present invention provides a chromatography column, a scale is provided on the column, to facilitate the operator to grasp the packing height and the sample volume; silica provided three cylinder inner wall of the tube, to facilitate a buffer, the sample liquid, the eluent other liquid flow from the cylinder walls, to avoid splashing. 本发明结构简单,操作方便,蛋白的纯化效果好。 The present invention is of simple structure, convenient operation, good purification effect of the protein.

Description

一种用于蛋白纯化的层析柱 Chromatography column for the purification of proteins

技术领域 FIELD

[0001]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。 [0001] The present invention pertains to a biological purification device, particularly relates to a column for purification of proteins.

背景技术 Background technique

[0002]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。 [0002] Currently, the compositions of the separation and purification of the compounds, natural products and medicine, mainly by column chromatography using a column, achieved by the chromatography column. 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。 Traditional column chromatography mixing the compound is placed on top of a glass column bed, the mixed solvent composition of the eluent (mobile phase) under gravity, through the dielectric material containing adsorbent (stationary phase) in a vertical glass column, to give each pure isolated product (pure), its principle is: the compound due to the mixing of the components in affinity for the column stationary phase and the mobile phase difference, so selecting a suitable stationary phase and the mobile phase mixing of the compounds according to the nature , can achieve the purpose of separation in which the components as much as possible, the separation of the product outlet in the column are collected in each collection bottle, then collected to identify each flask to collect liquid chromatography, according to the identification results combined and concentrated isotropic liquid chromatography, to obtain the respective pure product.

[0003]常见层析柱根据压力不同可分为常压柱、中压柱和高压柱。 [0003] Depending on a common column can be divided into atmospheric pressure column, medium pressure column and the higher pressure 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 used, their use is often different column materials according to the needs, 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 chromatography column and used in particular, often including a cylinder, a piston, and some further comprises barbs for fixation fluid like a ball at an upper end of the cylinder. 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迀移速度,然后根据迀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。 Current common glass chromatography column following defects: on the one hand, a glass column packed in packing and filling the sample substances to be separated, often using different fillers and different materials according to different fillers, the proportion of the sample, in order to facilitate to achieve the most while good separation effect savings filler, and the filler proportion of the sample is generally a volume ratio with reference to, but is not currently used scale glass columns, often require the operator to visually or measured empirically, other very troublesome metrology tool; the other hand, in the chromatographic process, often need to know the speed of the sample separation medium that is adjusted fillers Gan shift speed, and then the effluent shift speed according to Gan, to facilitate saving time while achieving optimal chromatographic separation effect, but at present no scale chromatography column, often at different time points to the location of a mark with a marker ribbon chromatography, such an approach is not accurate enough, while the marker token may be marked as eluent leakage when added It has been washed away, to be marked again, very convenient.

[0004]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。 [0004] In addition, the current column during injection, the sample solution flow directly into the cylinder from the middle of the top of the column, within the column packing is likely to cause splashing on the inner walls of the 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 fillers are often relatively expensive, splashing on the inner wall of the filler is not fully utilized in chromatography, chromatography impact effect.

发明内容 SUMMARY

[0005]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: [0005] In order to solve the above problems, the present invention provides a chromatography column for purifying the protein, the following technical solution:

[0006] —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0.1〜0.45μmD [0006] - species chromatography for purification of proteins, including stigma and the post, and the stud is threaded cylinder, wherein said top is provided with holes for stud plastic tube passes; the said cartridge transparent plastic material, provided with a uniform scale along the axial direction of the cylinder on the column; the column bottom of the plastic tube is put into a four-way pipe connector body, the end of the crossbar of the other three are connected to a silicone tube, the other end of the silicone tube contacts the inner wall of the cylinder; the cylinder bottom tapered outlet; an upper deck of the tapered outlet is provided, said frit pore size 0.1~0.45μmD

[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 connection of said cylinder stud and a sealing ring, the sealing ring is a silicone material.

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

[0009] 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迀移速率,使整个层析过程更方便,提高分离效率和整个工作效率。 [0009] 1. The present invention provides a chromatography column, the column transparent plastic material, and a uniform scale provided on the column, thus provided, is conducive to convenient filler loading in the sample and sample chromatography column accurately grasp the sample packing ratio, by observing the position of the scale of a ribbon located at different time points, so as to facilitate grasp Gan substances to be separated in a chromatography column shift rate, the whole chromatography process easier, and improve the separation efficiency overall efficiency.

[0010] 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果O [0010] 2. The present invention provides a chromatography column, the column extending into the bottom of the plastic tube connected to the body of a crossbar, said crossbar end is connected to each of the other three a silicone tube, the silicone hose the other end contacts the inner wall of the cylinder, so arranged conducive buffer, the sample liquid, when an eluent from the top into the column, a stream along the sidewall of the pillar, the body without causing splashing of the liquid column, fillers cause splashing on the sidewalls of the pillar, the influence packing height, thus affecting protein purification effect O

[0011] 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0.1〜 [0011] 3. The present invention provides a chromatography column, the outlet is provided at an upper conical sieve, and the sieve aperture defining 0.1~

0.45μπι,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。 0.45μπι, when the filler so this will not be achieved with the purified protein, the loss from the tapered outlet with buffer, sample liquid eluant, but without affecting the buffer, the eluent from the sample liquid and the cone shaped outlet outflow effect.

[0012] 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。 [0012] 4. The chromatography column of the present invention provides, at the junction with the sealing ring and the post stigma, stigma and facilitate hermetic cylinder, to avoid buffer, sample liquid from the eluent gap at both outflow, resulting in waste.

[0013] 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。 [0013] 5. The chromatography column of the present invention provides a simple structure, easy operation, good purification effect of the protein.

附图说明 BRIEF DESCRIPTION

[0014]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表示刻度;5表示筛板;6表示硅胶软管;7表示柱体。 [0014] FIG. 1 is a schematic structural diagram of the present invention; wherein, I represents stigma; 2 represents a plastic tube; 3 denotes a conical outlet; 4 represents a scale; sieve represents 5; 6 denotes a silicone tube; 7 represents a column.

具体实施方式 detailed description

[0015]下面结合附图和实施例对本发明作进一步说明。 Drawings and embodiments of the present invention will be further described [0015] below in conjunction. 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 embodiments, the present invention is capable of other embodiments, where the use of equivalent replacement or equivalent transformation formed technical solutions all fall within the scope of the invention claimed.

[0016]如图1所示,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0.1〜0.45μηι。 [0016] As shown, a process for purifying a protein chromatography column, including the column 7 and stigma I, column I and the stud 7 is screwed, characterized in that the stigma of top I 2 is provided with through holes for plastic tubes; and the cylinder 7 made of transparent plastic material, provided along the axial direction of the cylinder 4 on the scale of uniform the cartridge 7; 2 of the plastic tube 7 extends into the cartridge connecting a bottom of the crossbar, the crossbar of the other three terminals are connected to a silicone tube 6, the other end of the hose silica gel column contact the inner wall 6 of 7; and a bottom of the column 7 3 is a tapered outlet; upper portion of the tapered outlet 3 provided 5 sieve, the sieve aperture 5 is 0.1~0.45μηι.

[0017]作为优选,在柱头I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。 [0017] Advantageously, a sealing ring is not marked out in the figure, for the sealing action of the sealing ring is a silicone material, better sealing effect in a cylinder and connected at the stigma I 7.

Claims (1)

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0.1-0.45ym; 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。 A chromatography column for the purification of proteins, including stigma (I) and the post (7), said stud (I) and the cartridge (7) is screwed, characterized in that said stud (I ) is provided for the top of the plastic tube (2) through the through-holes; said cartridge (7) made of transparent plastic material, said body (7) axially along the cylinder is provided with a uniform scale (4) ; bottom (2) extends into the cartridge (7) within the plastic tube connected to a crossbar, the other three ends of the crossbar are connected to a silicone tube (6), the silicone tube (6 ) the other end of the inner wall of the cylinder (7) of the contact; said cartridge (7) is tapered at the bottom an outlet (3); the upper portion of the tapered outlet (3) disposed deck (5), the sieve (5) having a pore size 0.1-0.45ym; the stigma (I) and the post (7) is connected at a sealing ring, the sealing ring is a silicone material.
CN 201310644051 2013-12-05 2013-12-05 Chromatography column for the purification of proteins CN103691154B (en)

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Citations (6)

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Publication number Priority date Publication date Assignee Title
US6280616B1 (en) * 1995-02-21 2001-08-28 Pharmacia Biotech Ab Column for chromatography
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CN201379989Y (en) * 2009-01-05 2010-01-13 栗云天 Chromatographic column assembly
CN202237384U (en) * 2011-09-19 2012-05-30 何慧 Chromatographic column for laboratory
CN202410236U (en) * 2011-12-28 2012-09-05 上海师范大学 Detachable chromatographic column with scales
CN203678008U (en) * 2013-12-05 2014-07-02 成都雅途生物技术有限公司 Chromatographic column used for protein purification

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6280616B1 (en) * 1995-02-21 2001-08-28 Pharmacia Biotech Ab Column for chromatography
CN201223741Y (en) * 2008-07-07 2009-04-22 江苏汉邦科技有限公司 Small-sized simulated moving bed chromatography device
CN201379989Y (en) * 2009-01-05 2010-01-13 栗云天 Chromatographic column assembly
CN202237384U (en) * 2011-09-19 2012-05-30 何慧 Chromatographic column for laboratory
CN202410236U (en) * 2011-12-28 2012-09-05 上海师范大学 Detachable chromatographic column with scales
CN203678008U (en) * 2013-12-05 2014-07-02 成都雅途生物技术有限公司 Chromatographic column used for protein purification

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