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Publication numberCN103691154 B
Publication typeGrant
Application numberCN 201310644051
Publication date11 May 2016
Filing date5 Dec 2013
Priority date5 Dec 2013
Also published asCN103691154A
Publication number201310644051.3, CN 103691154 B, CN 103691154B, CN 201310644051, CN-B-103691154, CN103691154 B, CN103691154B, 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
一种用于蛋白纯化的层析柱 Column for the purification of proteins translated from Chinese
CN 103691154 B
Abstract  translated from Chinese
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 method for protein purification column, which belongs to the field of bio-purification equipment. 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, the column is provided on the scale to facilitate the operator of the packing height and the injection volume control; and the cylinder wall three silicone hose set for easy buffer, sample solution, the eluent and 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 simple in structure, easy operation, good purification effect protein.
Claims(1)  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0.1-0.45ym; 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 column for the purification of proteins, including the stigma (I) and the cylinder (7), the stigma (I) and the cylinder (7) is screwed, characterized in that the stigma (I ) is provided for the top plastic tube (2) through the through hole; said cylinder (7) with a transparent plastic material, said cylinder (7) along the axial direction of the cylinder is provided with a uniform scale (4) ; bottom (2) extending into the cylinder (7) within the plastic tube connected to a crossbar, the crossbar of the other three terminals are connected to a silicone hose (6), the silicone hoses (6 ) and the inner wall of the other end of the cylinder (7) is in contact; the bottom of the 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 0.1-0.45ym; 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 Column for the purification of proteins

技术领域 TECHNICAL 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 is a biological purification device, particularly relates to a column for the 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 separation and purification of compositions of the compounds, natural products and traditional Chinese medicine, primarily by column chromatography using a column by column chromatography process to achieve. 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 the action of gravity, through the dielectric material containing adsorbent (stationary phase) vertical glass column, isolated individual purer product (pure), the principle is: As the mixing of the components of compound affinity column fixed and mobile phases are different, so choose the appropriate fixed and mobile phases depending on the nature of the mixed compound can achieve the purpose of which will be separated as much as possible of the components, the isolated products were collected at the column outlet to each collection bottle, and then identify each collection vial collected liquid chromatography, based on the identification results combined and concentrated isotropic liquid chromatography to give each 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] According to common column pressure can be divided into atmospheric column, medium pressure column and 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 used most often is his use of different materials column as needed, filling different fillers to achieve the 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 was especially popular, often including cylinder, piston, and some also includes a column for the upper end of barbs and other fixed dosing 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。 Existing common glass column less than the following: on the one hand, a glass column packed in packing and loading a sample substances to be separated, it is often different depending on packing and packing of different substances, the proportion of the sample, in order to facilitate reaching the most At the same time good separation effect savings filler, and filler proportion of the sample usually by reference to volume ratio, but there is no common column glass scale, often require the operator to visually or empirically measured, very troublesome to use other metrics tool; the other hand, in the chromatographic process, often need to know the sample separation medium, the filler Gan motion speed, then the effluent according to Gan traverse speed regulation, in order to facilitate optimum chromatographic separation efficiency while saving time, but now the column does not scale, often at different time points to a location with a marker chromatography ribbon marker, such an approach is not accurate enough, while marking pen mark of identification may be added as the eluent leakage when It was 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 flows into the cylinder from the top of the middle column directly, likely to cause the inner wall of the column packing body splash to 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 fillers are often more expensive, splashing onto the inner wall of the filler material have not been fully utilized in chromatography, chromatography affect results.

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] To solve the above problems, the present invention provides a method for protein purification by column chromatography, using 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] - the kind used for protein purification column, including the column and stigma, the stigma and the cylinder is screwed, characterized in that the top of the stigma of a through-hole for a plastic tube through; the said cylinder with a transparent plastic material, provided with a uniform scale along the axial direction on the cylinder barrel; the bottom of the plastic tube is put into the body of the column connecting a crossbar, the crossbar of the other three end a silicone hoses are connected, the other end of the silicone tube in contact with the inner wall of the cylinder; the bottom of the cylinder is tapered outlet; the upper part of the conical outlet is provided sieve, said sieve 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 the stigma and the cylinder is provided with seals, the seals of silicone material.

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

[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 set a uniform scale on the cylinder, so arranged beneficial when spotted on a sample column filler loading and convenience filling sample accurately grasp proportion, by observing a ribbon at different points in time where the scale position, making it easy to grasp Gan substances to be separated in a chromatography column shift rate, so that the whole chromatography process easier, and to 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, a plastic tube is put into the body of the bottom of the column connecting a crossbar, the crossbar of the other three terminals are connected to a silicone hose, the Silicone hose and the other end in contact with the inner wall of the cylinder, so setting conducive buffer, sample solution, eluent when entering from the top of the column, is shed along the sidewall of the cylinder, the cylinder body without causing splashing liquid, cause packing splash on the side wall of the cylinder, the impact of packing height, thus affecting the 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, set at the upper tapered outlet sieve, and the sieve aperture is defined 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, so that we can achieve such a filler protein purification will not end with buffer, sample solution, together with the eluent from the tapered outlet drain, but will not affect the buffer, the sample solution and the eluent from the cone outflow shaped outlet 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 present invention provides a chromatography column provided at the junction of the stigma and cylinder seals, cylinder facilitates stigma and sealed to avoid buffer, sample solution, the eluent from the crevice between the two 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 present invention provides a chromatography column, simple structure, easy operation, good purification effect 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] Figure 1 a schematic view of the structure of the present invention; wherein, I represents stigma; 2 represents a plastic pipe; 3 showing the tapered outlet; 4 shows scale; sieve represents 5; 6 represents silicone hose; 7 represents 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。 [0015] below in conjunction with the accompanying drawings and the embodiment of the present invention will be further described. 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 equivalent replacement or equivalent transformed form aspect, 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] Figure 1, for protein purification by column chromatography, including column and stigma I 7, I and the stigma is threaded cylinder 7, characterized in that the top of stigma I It has 2 holes for plastic pipe through; the cylinder 7 made of transparent plastic material, the cylinder 7 along the cylinder, is provided with a uniform scale 4; the plastic tube inserted into the cylinder 2 7 the bottom of the inner tube connected to a four-way, four-way tube of the other three terminals are connected to a silicone hose 6, the other end of the silicone hose with an inner wall of the cylinder 6 in contact 7; the bottom of the column 7 tapered outlet 3; upper part of the conical outlet 3 of 5 sieve set, the aperture sieve 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, the stigma I and column junction ring 7 is provided in the figure is not marked out for the seal, the seal is silicone material, seal better.

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Classifications
International ClassificationB01D15/22, C07K1/16
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