CN107045066A - Immunoblotting reel - Google Patents

Immunoblotting reel Download PDF

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Publication number
CN107045066A
CN107045066A CN 201710190186 CN201710190186A CN107045066A CN 107045066 A CN107045066 A CN 107045066A CN 201710190186 CN201710190186 CN 201710190186 CN 201710190186 A CN201710190186 A CN 201710190186A CN 107045066 A CN107045066 A CN 107045066A
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immunoblotting
strip
region
reel
immunoblot
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CN 201710190186
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Chinese (zh)
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黄世海
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武汉优视科技有限公司
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by the preceding groups
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803General methods of protein analysis not limited to specific proteins or families of proteins

Abstract

The invention discloses an immunoblotting reel. The external surface of the periphery of the immunoblotting reel is provided with a plurality of parallel strip-shaped hydrophilic zones which are separated from each by hydrophobic zones; the strip-shaped hydrophilic zones are arranged to be vertical to a center axis of the immunoblotting reel; each of the strip-shaped hydrophilic zones is provided with one or a plurality of combination zones along the length direction; each of the strip-shaped hydrophilic zones is used for combination with different solutions to be detected, and is used for independent immunoblotting detection of target antibodies in the different solutions to be detected; the combination zones are provided with target antigens which are used for combination with the target antibodies in the solutions to be detected. The immunoblotting reel is provided with the plurality of parallel strip-shaped hydrophilic zones which are separated from each by the hydrophobic zones, combination with the target antibodies is realized under the absorption foce of the hydrophilic zones, so that reagent is saved, and the immunoblotting reel is suitable to be used for trace detection.

Description

一种免疫印迹卷轴 An immune blot reel

技术领域 FIELD

[0001] 本发明属于蛋白质分析领域,更具体地,涉及一种免疫印迹卷轴。 [0001] The present invention belongs to the field of protein analysis and, more particularly, to a reel immunoblotting.

背景技术 Background technique

[0002] 免疫印迹(Western Blot)技术是将蛋白质转移到膜上,然后利用抗体进行检测的技术。 [0002] Immunoblotting (Western Blot) techniques is transferred to a membrane protein, and then use technology for detecting antibodies. 免疫印迹常用于鉴定某种蛋白,并能对蛋白进行定性和半定量分析。 Immunoblotting used for the identification of a protein, and proteins capable of qualitative and semi-quantitative analysis. 结合化学发光检测,可以同时比较多个样品中同种蛋白的表达量差异。 Binding chemiluminescent detection, you can compare multiple samples isoform differences in protein expression. 由于免疫印迹具有SDS-PAGE的高分辨力和固相免疫测定的高特异性和敏感性,现己成为蛋白分析的一种常规技术。 Since SDS-PAGE immunoblotting with high resolution and a solid phase immunoassay with high specificity and sensitivity, it has now become a routine technique for protein analysis.

[0003] 为了实现高效率的自动化分析,各种各样的免疫印迹分析仪应需而生(专利文献CN103616526A)。 [0003] In order to achieve high efficiency of automatic analysis, a variety of immunoblot analysis instrument should be born (Patent Document CN103616526A). 然而,上述免疫印迹分析仪通常利用多个试剂瓶或反应槽进行多个待测样本的平行检测,同时利用摇床将试剂瓶进行振荡,以加快反应速度。 However, the above western blot analyzer reagent bottles typically utilize a plurality of reaction vessel or parallel detection of a plurality of samples to be tested, while the use of the reagent bottle shaker was shaken to accelerate the reaction rate. 因此其具有以下缺点: 一、由于集成了用于振荡试剂瓶的摇床,使得整个分析仪结构较为复杂,进而提高了制造成本;二、不同试剂瓶的反应条件略有差异,从而影响免疫印迹的准确性;三、薄膜需要从试剂瓶中取出后进行干燥才可进行检测,从而无法实现原位检测。 Thus it has the following disadvantages: First, since the integrated reagent for oscillating shaker bottle, so that the entire structure of the analyzer is complicated, thereby increasing manufacturing cost; second, different reaction conditions are slightly different reagent bottle, thus affecting immunoblotting accuracy; Third, the film needs to be removed from the vial and dried before detection, and thus can not achieve in situ detection.

发明内容 SUMMARY

[0004] 针对现有技术的以上缺陷或改进需求,本发明提供了一种免疫印迹卷轴,其目的在于简化免疫印迹自动检测系统的结构、同时实现原位检测。 [0004] To solve the above drawbacks of the prior art or the need for improvement, the present invention provides an immunoblot reel, and its object is to simplify the configuration immunoblotting automatic detection system, while achieving in situ detection.

[0005] 为实现上述目的,按照本发明的一个方面,提供了一种免疫印迹的自动检测系统, 包括免疫印迹卷轴、加液装置以及废液收集装置;所述免疫印迹卷轴的周向的外表面具有以疏水区域间隔的多个平行的条状亲水区域,所述条状亲水区域垂直于所述免疫印迹卷轴的中心轴,所述条状亲水区域沿其长度方向具有一个或多个结合区,其中,每个所述条状亲水区域分别用于结合不同的待测溶液,从而独立地进行不同的待测溶液中的目标抗体的免疫印迹检测,所述结合区上具有目标抗原,用于与待测溶液中的目标抗体结合;所述加液装置用于向条状亲水区域添加反应液,所述废液收集装置用于收集多余的反应液;所述反应液包括待测溶液、缓冲液、二抗溶液或显色溶液。 [0005] To achieve the above object, according to one aspect of the present invention, there is provided a system for automatic detection of the Western blot, immunoblot comprises a reel, the waste liquid collecting device and a dosing device; immunoblot said spool outer circumferential surface having a plurality of parallel strip-shaped hydrophilic areas at spaced hydrophobic region, a hydrophilic region of the strip to the central axis perpendicular immunoblot reel, the strip having one or more hydrophilic regions along its length binding region, wherein each of said strip-shaped hydrophilic areas are different solutions to be tested for binding to independently perform immunoblotting antibody of different solutions to be tested in the detection of a target binding region of the antigens, used in combination with the antibody in the test solution; the dosing means for adding the reaction solution to a hydrophilic region of the strip, the waste liquid collecting means for collecting excess reaction liquid; and the reaction solution comprising test solution, a buffer, a secondary antibody solution or a developing solution.

[0006] 按照本发明的另一个方面,还提供了一种用于上述自动检测系统的免疫印迹卷轴,所述免疫印迹卷轴的周向的外表面具有以疏水区域间隔的多个平行的条状亲水区域, 所述条状亲水区域垂直于所述免疫印迹卷轴的中心轴,所述条状亲水区域沿其长度方向具有一个或多个结合区,其中,每个所述条状亲水区域分别用于结合不同的待测溶液,从而独立地进行不同的待测溶液中的目标抗体的免疫印迹检测,所述结合区上具有目标抗原,用于与待测溶液中的目标抗体结合。 [0006] According to another aspect of the present invention there is also provided one of the above western blot detection system for automatic reels, the spool immunoblot circumferential outer surface having a plurality of parallel spaced strips at the hydrophobic region hydrophilic regions, hydrophilic regions of the strip perpendicular to the central axis of the spool immunoblotting, the strip-shaped region having one or more hydrophilic binding regions along its length, wherein each of said strip-like pro water areas are different solutions to be tested for binding to independently perform immunoblotting antibody of different solutions to be tested in the detection of the antigen having a binding region for binding to the antibody in the test solution .

[0007] 优选地,所述免疫印迹卷轴包括转轮以及覆盖于所述转轮周向的外表面的免疫印迹薄膜;所述转轮用于固定免疫印迹薄膜,所述免疫印迹薄膜具有以疏水区域间隔的多个平行的条状亲水区域。 [0007] Preferably, said spool comprises a wheel and immunoblotting covering the outer surface of the wheel circumferential direction of the immunoblot films; immunoblotting for fixing the reels immunoblot film, said film having a hydrophobic a plurality of parallel strip-like hydrophilic regions spaced areas.

[0008] 作为进一步优选地,所述免疫印迹薄膜粘贴于所述转轮周向的外表面。 [0008] As a further preferably, the film is bonded to immunoblotting outer surface of the wheel circumferential direction.

[0009] 作为进一步优选地,所述免疫印迹薄膜的底面以及所述转轮周向的外表面具有磁性涂层,所述免疫印迹薄膜通过磁力吸附于所述转轮周向的外表面。 [0009] As a further preferably, the film immunoblotting bottom surface and an outer surface of said wheel having a circumferential magnetic coating, the outer surface of the immunoblot films by magnetically attracting to the wheel circumferential direction.

[0010] 作为进一步优选地,所述转轮的外周设置有卡扣,所述卡扣将所述免疫印迹薄膜固定于所述转轮周向的外表面。 [0010] As a further preferably, the outer periphery of the wheel is provided with a buckle, the buckle immunoblotting the outer surface of the film is fixed to the wheel circumferential direction.

[0011] 优选地,所述免疫印迹卷轴的直径为2cm〜6cm,宽度为2cm〜6cm。 [0011] Preferably, the diameter of the reel immunoblotting 2cm~6cm, width 2cm~6cm.

[0012] 优选地,所述条状亲水区域的材料为硝酸纤维素、尼龙或聚偏氟乙烯。 [0012] Preferably, the hydrophilic region of the strip material is nitrocellulose, nylon or polyvinylidene fluoride.

[0013] 优选地,所述疏水区域具有硅烷基。 [0013] Preferably, the hydrophobic region having a silane group.

[0014] 作为进一步优选地,所述疏水区域的材料为聚二甲基娃氧烷或其衍生物。 [0014] As further preferred that the material of the hydrophobic region of the baby polydimethyl siloxane or derivative thereof.

[0015] 优选地,所述条状亲水区域以及疏水区域的宽度为1mm〜5mm。 Width [0015] Preferably, the strip-shaped hydrophilic region and a hydrophobic region is 1mm~5mm.

[0016] 优选地,所述条状亲水区域具有多个结合区,所述多个结合区包括实验区以及对照区。 [0016] Preferably, the hydrophilic region having a plurality of strip-shaped binding region, the binding region comprises a plurality of experimental area and control area.

[0017] 作为进一步优选地,所述对照区包括阳性对照区、阴性对照区或结合物对照区。 [0017] As a further preferably, the control zone comprises a positive control area, a negative control region or a control region in combination thereof.

[0018] 总体而言,通过本发明所构思的以上技术方案与现有技术相比,具有下列有益效果: [0018] In general, compared with the prior art by the above technical solution of the inventive concept, it has the following advantages:

[0019] 1、免疫印迹卷轴具有以疏水区域间隔的多个平行的条状亲水区域,通过亲水区域的吸附力以及水的表面张力与待测溶液中的目标抗体结合,通过疏水区域的隔离防止反应液漏出,仅需要毫升级以下的反应液即可完成免疫印迹检测,从而更加节省试剂,适用于痕量检测; [0019] 1, the reel having a plurality of strip immunoblot hydrophilic regions parallel spaced hydrophobic regions, hydrophilic regions joined by the suction force and the surface tension of the aqueous solution to be measured in the antibody, hydrophobic regions through isolation prevents leakage of the reaction liquid, need only milliliter or less to complete the reaction was detected by immunoblotting, thus saving more reagents for the detection of trace;

[0020] 2、多种待测溶液中的不同目标抗体可结合于同一免疫印迹卷轴的不同条状亲水区域的结合区上,从而更适于在完全相同的条件下进行反应,易于进行平行对照,提高了检测的准确率;同时简化了自动检测系统的结构,降低了生产成本; The binding regions of different stripe hydrophilic regions [0020] 2, a variety of different target test solution may bind to the same antibodies immunoblot reel, and thus more suitable for reaction under exactly the same conditions, is susceptible to parallel control, improve the accuracy of detection; while simplifying the structure of an automatic detection system, reduces the production cost;

[0021] 3、免疫印迹卷轴通过条状亲水区域的吸附力以及水的表面张力与反应液结合,从而使得所需的反应液的体积较少,简化了检测步骤,更适用于原位检测; [0021] 3, the suction force by immunoblotting reel strip and the surface tension of the hydrophilic region with water binding reaction liquid, so that the volume of the reaction solution is less required, simplifies the detection step, more suitable for in situ detection ;

[0022] 4、免疫印迹卷轴可由免疫印迹薄膜绕制而成,每次免疫印迹反应完成后,无需更换免疫印迹卷轴,只需撕去表层的免疫印迹薄膜即可再次进行免疫印迹反应,从而节省了反应时间,简化了操作步骤;同时该免疫印迹卷轴能减少储存空间。 [0022] 4, immunoblotting film reel can be wound from immunoblotting, western blot after each completion of the reaction, without changing the reel immunoblot, only tear film surface to immunoblot immunoblotting reaction again, thereby saving reaction time, simplified the process; a reel at the same time to reduce the immunoblot storage.

附图说明 BRIEF DESCRIPTION

[0023]图1是本发明自动检测系统正视图; [0023] FIG. 1 is a front view of an automatic detection system of the present invention, FIG;

[0024]图2是本发明免疫印迹卷轴的周向的外表面的展开图; [0024] FIG. 2 is a developed view of an outer circumferential surface of the spool of the present invention immunoblotting;

[0025]图3a为本发明免疫印迹卷轴所用的衬底膜的示意图; [0025] Figure 3a is a schematic diagram of the substrate film used in the immunoblot spool invention;

[0026]图3b为本发明免疫印迹卷轴制备过程中形成结合带示意图; [0026] FIG. 3b immunoblotting binding band diagram reel formed during the preparation of the present invention;

[0027]图3c为本发明免疫印迹卷轴制备过程中形成疏水区域示意图; [0027] FIG. 3c immunoblot schematic reel formed during the preparation of the present invention, the hydrophobic region;

[0028]图4为本发明将条状亲水薄膜平行贴制于疏水薄膜过程示意图; [0028] FIG. 4 of the present invention will be hydrophilic membrane strip affixed parallel to the hydrophobic membrane made of a schematic process;

[0029] [0029]

[0030] [0030]

[0031]图5为具有凹槽的疏水薄膜表面固定条状亲水薄膜示意图; [0031] FIG. 5 is a schematic view of a strip fixed hydrophobic film hydrophilic membrane having a surface of the recess;

[0032]在所有附图中,相同的附图标记用来表示相同的元件或结构,其中:11-疏水区域, 12-条状亲水区域,13-结合区,15-免疫印迹薄膜,17-转轮,2-加液装置,4-信号获取装置, 41 -图像传感器,42-光源,5-废液收集装置。 [0032] Throughout the drawings, like reference numerals are used to designate the same elements or structures, wherein: 11- hydrophobic region, a hydrophilic region strip 12-, 13- binding region, immunoblot films 15, 17 - wheel, 2 dosing device, 4 signal acquisition device, 41 - an image sensor, a light source 42-, 5-waste collection device.

具体实施方式 detailed description

[0033]为了使本发明的目的、技术方案及优点更加清楚明白,以下结合附图及实施例,对本发明进行进一步详细说明。 [0033] To make the objectives, technical solutions and advantages of the present invention will become more apparent hereinafter in conjunction with the accompanying drawings and embodiments of the present invention will be further described in detail. 应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。 It should be understood that the specific embodiments described herein are only intended to illustrate the present invention and are not intended to limit the present invention. 此外,下面所描述的本发明各个实施方式中所涉及到的技术特征只要彼此之间未构成冲突就可以相互组合。 Moreover, various embodiments of the invention described below involved the technical features as long as no conflict with one another can be configured in combination with each other.

[0034]如图1所示,本发明公开了一种免疫印迹的自动检测系统,包括免疫印迹卷轴、力口液装置2、信号获取装置4以及废液收集装置5等; [0034] As shown in FIG. 1, the present invention discloses a system for automatic detection of the Western blot, immunoblot comprises a reel, oral fluid power means 2, the signal acquisition device 4 and the waste liquid collection apparatus 5 and the like;

[0035]如图2所示,所述免疫印迹卷轴的周向的外表面具有以疏水区域11间隔的多个平行的条状亲水区域12,疏水区域11以及条状亲水区域12的宽度为1mm〜5mm,在便于做免疫印迹测试的同时,可保证各条状亲水区域12完全间隔,同时有效利用免疫印迹卷轴的空间, 所述条状亲水区域12垂直于所述卷轴的中心轴,其具有一个或多个结合区13,其周向的外表面的展开图如图2所示;当所述条状亲水区域12包括多个结合区12时,结合区12可分为实验区以及对照区,实验区用于与目标蛋白结合,而对照区根据其功能,可分为阳性对照区、 阴性对照区或结合物对照区;其中,阳性对照区通常含有能与待测溶液中除目标抗体以外的其它成分结合的抗原,例如,当以血液为待测溶液时,阳性对照区的目标抗原则可采用血清抗体,阳性对照区显色证明免疫印迹薄膜15未变质,且待 Width of the outer surface of the [0035] 2, the circumferential immunoblot spool having a plurality of parallel spaced hydrophobic region 11 of the strip 12 hydrophilic regions, hydrophobic regions 11 and hydrophilic regions 12 of the stripe is 1mm~5mm, while facilitating the immunoblot test, the hydrophilic region can ensure that each stripe 12 is completely spaced, while effectively by immunoblotting reel space 12 perpendicular to the strip-shaped hydrophilic region of the central spool a shaft having one or more binding regions 13, which expand the outer surface of a circumferential direction shown in Figure 2; when the strip-shaped region 12 includes a plurality of hydrophilic binding regions 12, 12 can be divided into binding region experimental area and control area, experimental area for binding to the target protein, according to their function and the control zone, the control zone can be divided into positive, negative control region or a control region in combination thereof; wherein the positive control area and typically contains a solution can be measured in addition to the other components of the antibody bound antigen, e.g., when the blood into the test solution to the target region of the positive control serum antibody can be an anti-principle, the positive control area color proof film 15 is not deteriorated immunoblotting, Judai 测溶液种类无误;而阴性对照区可涂覆有未标记的二抗,使得在标准条件下阴性对照区无法显色,若其显色,证实可能有免疫印迹薄膜15变质等异常情况,应该更换免疫印迹薄膜15而重新测定;而结合物对照区的目标抗原为IgG结合物、IgM结合物或IgA结合物,由于IgG、IgM和IgA为常见的免疫球蛋白, 结合物对照区的存在可指示目标抗体是否已经成功结合。 Correct type of Solution; and negative control zone may be coated with an unlabeled secondary antibody, negative control region such that under standard conditions, the color can not, if its color, there may be confirmed by immunoblotting film deterioration and other anomalies 15, should be replaced immunoblotting film 15 and the remeasurement; the antigen was incorporated in the control area as IgG conjugate, IgM or IgA bound conjugate was due to the IgG, IgM and IgA immunoglobulin is common, the presence of binding may indicate that the control area thereof whether the target antibody has been successfully combined.

[0036] 所述加液装置2具有与条状亲水区域12—一对应的多个加液通道,所述加液通道的出水口与对应的条状亲水区域12相对设置,距离为0.1mm以上(当加液通道设置于条状亲水区域12的侧面时,为了保证反应液不泄露,距离应为0.1mm〜2mm,而当加液通道设置于条状亲水区域12的上方时,距离可以更远),所述免疫印迹卷轴底部的条状亲水区域12连接废液收集装置5的进水口;同时,还可设置与条状亲水区域12相对的超声探头,以发出超声信号,以加快免疫印迹卷轴表面的反应速度。 [0036] The dosing device 2 having a plurality of dosing channels and the strip corresponding to a hydrophilic region 12-, the dosing passage outlet and the strip-shaped hydrophilic areas 12 corresponding to opposite from 0.1 mm or more (when the dosing passage on the side surface 12 of the strip-shaped hydrophilic zone, in order to ensure that the reaction liquid does not leak, the distance should be 0.1mm~2mm, when the dosing passage is provided above the strip 12 of hydrophilic region , the distance can be longer), the bottom of the strip immunoblot spool 12 connected to the water inlet hydrophilic region waste collection device 5; the same time, the hydrophilic region can also be provided with a strip 12 opposite ultrasound probe to emit ultrasound signal to accelerate the reaction rate of the surface of the reel immunoblotting.

[0037] 信号分析装置包括光源42以及图像传感器41,用于获取免疫印迹的检测信号;根据二抗溶液中二抗的标记,所述光源42可选用激光光源42或可见光光源42 (例如常用标记异硫氰酸荧光素的最大吸收光波长为490nm〜495nm,四乙基罗丹明的最大吸收光波长为570nm),用于向免疫印迹薄膜15方向发出可见光或激光,在免疫印迹薄膜15表面形成免疫印迹的检测信号,无论是可见光或激光,光源42需要光线均匀;光源42的形状优选为长形, 宽度与免疫印迹卷轴的高度相当,其与条状亲水区域12的方向垂直,但与免疫印迹卷轴的中心轴平行,可以保证投射到每个条状亲水区域12同一位置的光线均匀,以免出现对比误差;所述图像传感器41用于获取免疫印迹的检测信号。 [0037] The signal analyzing apparatus includes a light source 42 and an image sensor 41, for obtaining a detection signal immunoblot; The secondary antibody labeled secondary antibody solution, the light source 42 is a laser light source 42 can be selected or visible light source 42 (e.g., common numerals the maximum absorption wavelength of light for fluorescein isothiocyanate 490nm~495nm, tetraethyl rhodamine maximum absorption wavelength of light is 570nm), a thin film direction 15 to emit visible light or laser immunoblotting, western blot film 15 is formed on the surface immunoblot detection signal, whether visible or laser light source 42 requires a uniform light; a light source 42 is preferably of elongate shape, a width and a considerable height immunoblotting spool which is perpendicular to the direction of the hydrophilic stripe region 12, but immunoblotting spool parallel to the axis, each strip can be projected to ensure uniform light hydrophilic region 12 in the same position, in order to avoid comparison error; the image sensor 41 for acquiring immunoblot detection signal.

[0038] 所述免疫印迹卷轴的直径为2cm〜6cm,宽度为2cm〜6cm;其中,所述条状亲水区域12的材料为硝酸纤维素、尼龙或聚偏氟乙烯,以便结合目标抗原,所述疏水区域11通常为具有硅烷基的有机疏水聚合物,同时还可能携带有硅氧基或硅羟基,如聚二甲基硅氧烷或其衍生物。 [0038] The diameter of spool immunoblot 2cm~6cm, width 2cm~6cm; wherein the hydrophilic material of the strip-shaped region 12 of nitrocellulose, nylon or polyvinylidene fluoride in order to bind the target antigen, the hydrophobic region 11 is generally a hydrophobic organic polymer having a silane group and may also carry siloxy or silanol groups, such as polydimethylsiloxane or a derivative thereof.

[0039]如图1所示,免疫印迹卷轴可包括转轮17以及覆盖于所述转轮17周向的外表面的免疫印迹薄膜15,免疫印迹薄膜15具有以疏水区域11间隔的多个平行的条状亲水区域12, 其通过真空吸附、粘贴或卡扣等方式固定于所述转轮17上,所述转轮17用于带动所述免疫印迹薄膜15绕免疫印迹卷轴的中心轴旋转。 [0039] 1, immunoblotting spool 17 may include a wheel covering the outer surface of the periphery 17 of the wheel 15 film immunoblotting, western blot plurality of film 15 having a hydrophobic region 11 in parallel spaced hydrophilic region of the strip 12, and the like by vacuum suction, adhesive, or snap-fixed to the wheel 17, the center of the rotor shaft 17 for driving the film 15 around the western blot immunoblot rotation of the spool .

[0040]可应用于本发明的免疫印迹卷轴的免疫印迹薄膜15的制备方法大致分为两种: [0041]第一种:在衬底膜上结合目标抗原,以形成与目标抗原的数量相同的结合带,所述衬底膜的材料为硝酸纤维素、尼龙或聚偏氟乙烯,如图3所示;用疏水涂料在衬底膜上形成疏水区域11,以将衬底膜分隔为多个条状亲水区域12,同时将结合带分隔为与结合带数量相同的结合区13,使得每个条状亲水区域12都具有与目标抗原的数量相同的结合区13;该疏水涂料需要无毒,且在37度以下即可干燥,以免影响目标抗原的活性,如专利文件CN201380057250中的疏水涂层。 [0040] The present invention can be applied to immunoblotting Immunoblot spool 15 of the film production method roughly divided into two types: [0041] The first: the antigen binding substrate film, formed in the same amount of antigen combined with the substrate material is a nitrocellulose membrane, nylon or polyvinylidene fluoride, shown in Figure 3; hydrophobic region 11 formed in the substrate with a hydrophobic coating film to the base film is a multi-partition a hydrophilic region strip 12, while the partition is combined with the same amount of the binding band connecting region 13 so that each strip-like hydrophilic region 12 has the same number of the target antigen binding region 13; the hydrophobic coatings require non-toxic, and can be dried at 37 degrees or less, so as not to affect the activity of the target antigen, such as a hydrophobic coating in Patent document CN201380057250.

[0042]第二种:将具有结合区13的条状亲水薄膜平行贴制于疏水薄膜上,使得所述疏水薄膜上具有与多个条状亲水薄膜形成的条状亲水区域12,所述条状亲水区域12以疏水区域11相间隔,如图4所示;所述疏水薄膜表面可以设置有与条状亲水薄膜相配合的凹槽,以于与条状亲水薄膜结合并固定,使得条状亲水区域12所在的表面低于疏水所在的表面,这样可进一步保证条状亲水区域12之间的反应液不会互相影响混合,如图5所示。 [0042] The second: a strip having a binding region attached hydrophilic membrane 13 made parallel to the hydrophobic membrane, such that the hydrophobic film has a hydrophilic stripe region 12 is formed with a plurality of strip-shaped hydrophilic membrane, the hydrophilic region 12 to strip 11 spaced hydrophobic region, shown in Figure 4; said hydrophobic film surface may be provided with grooves cooperating with the hydrophilic membrane strip to the strip in combination with the hydrophilic film and fixed, so that the hydrophilic area of ​​the surface strip 12 is located below the surface where the hydrophobic, so that the reaction solution can be further ensured between the strip-shaped region 12 does not affect the hydrophilic mixed with each other, as shown in FIG.

[0043]上述自动检测系统可应用于免疫印迹检测,以间接法进行化学发光检测为例,具体包括以下步骤: [0043] The automatic detection system may be applied Western blot, indirect chemiluminescence detection method as an example, includes the following steps:

[0044] S1 •将多种待测溶液通过加液管以添加于不同的条状亲水区域12表面,使待测溶液中的目标抗体与结合带上的目标抗原完全结合,同时,可以利用与条状亲水区域12相对设置的超声探头6,以加快免疫印迹卷轴表面的反应速度;旋转电机在此时处于工作状态, 使得免疫印迹卷轴以Ocm/s〜15cm/s的线速度旋转30min〜60min;同时仍可以将待测溶液通过加液管缓慢添加于条状亲水区域12表面,以防液体蒸发,以保持条状亲水区域12表面上的液面高度为0.1mm〜1mm;在此情况下,经计算,即使将加液管中残留的待测溶液计算在内,所需的待测溶液的体积也仅需数十微升至毫升级; [0044] S1 • the plurality of the test solution was added by a dosing tube 12 to a different stripe hydrophilic surface region, the target antibody in the test solution fully integrated with the antigen binding belt, while, can be utilized the ultrasound probe 12 and the strip disposed opposite the hydrophilic region 6, in order to accelerate the reaction speed of the reel immunoblotting surface; rotating electrical machine is in operation at this time, so that the line speed of the reel immunoblotting Ocm / s~15cm / s rotation 30min ~60min; while still can be tested by adding the solution slowly added to the liquid pipe surface hydrophilic stripe region 12, to prevent the evaporation of a liquid to keep the liquid level on the surface of the strip-like hydrophilic region 12 is 0.1mm~1mm; in this case, by calculation, even if the test solution is added the calculated remaining in the inner tubing, the required volume of solution to be measured is also only several tens of microliters to milliliter;

[0045] S2.在免疫印迹卷轴的底部可设置集液管,集液管的一端距条状亲水区域的距离为0 • 1mm〜2mm,另一端连接真空栗,在反应完成后,真空栗通过集液管抽吸条状亲水区域12 表面多余的待测溶液,然后用真空泵抽吸使条状亲水区域12的表面干燥,或通过干燥装置辅助使之干燥; [0045] S2. Immunoblot spool may be provided at the bottom of the collecting channel, the distance from one end of the strip-shaped hydrophilic areas of the header pipes is 0 • 1mm~2mm, and the other end connected to a vacuum Li, after completion of the reaction, vacuum Li a suction tube strip through the header area of ​​the hydrophilic surface 12 to be measured excess solution, and then with a vacuum pump so that the surface 12 of the strip-shaped hydrophilic areas drying, or by a secondary drying apparatus and dried;

[0046] S3•以S1-S2.中同样的方式,以缓冲液替代原加液管中的待测溶液,将免疫印迹薄膜15表面的待测溶液冲洗约5min〜lOmin;由于缓冲液通常为PBS等成本较低的溶液,相对于待测溶液,可以设置为超过10ml/min以上的流量,而添加较多体积; [0046] S3 • S1-S2 in the same manner, the buffer instead of the solution to the original test tube was added, and the solution was tested by immunoblotting film surface 15 approximately flush 5min~lOmin;. Since the buffer solution is usually lower cost solution, such as PBS, with respect to the test solution, may be provided or more than 10ml / min flow rate, the more volume is added;

[0047] S4•以S1-S2.中同样的方式,以二抗溶液替代原加液管中的缓冲液,使得二抗完全与目标抗体结合; [0047] S4 • to S1-S2 in the same manner, the secondary antibody solution to replace the original buffer was added to the tube, so that the secondary antibody binding to the target antibody completely.;

[0048] S5•以S3.中同样的方式,以缓冲液替代原加液管中的二抗溶液,将免疫印迹薄膜15表面的二抗溶液冲洗干净; . [0048] S5 • S3 in the same manner, in order to replace the original buffer tube Dosing secondary antibody solution, the surface 15 of the film immunoblotting secondary antibody solution rinse;

[0049] S6•以S1-S2.中同样的方式,以显色溶液替代原加液管中的缓冲液,使得标记过的二抗发光; . [0049] S6 • to S1-S2 in the same manner, instead of the solution for color development buffer was added to the original tube, so that the labeled secondary antibody emission;

[0050] S7.以S3 •中同样的方式,以缓冲液替代原加液管中的显色溶液,将免疫印迹薄膜15表面的显色溶液冲洗干净; . [0050] S7 in the same manner to S3 •, to replace the buffer solution color original dosing tube, the surface 15 of the film immunoblotting color rinse solution;

[0051] S8•获得免疫印迹检测结果。 [0051] S8 • obtain immunoblot test results.

[0052] 本领域的技术人员容易理解,以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。 [0052] Those skilled in the art will readily appreciate, the above-described preferred embodiment of the present invention only but are not intended to limit the present invention, any modifications within the spirit and principle of the present invention, equivalent substitutions, and improvements, etc., should be included within the scope of the present invention.

Claims (9)

  1. 1. 一种免疫印迹卷轴,其特征在于,所述免疫印迹卷轴的周向的外表面具有以疏水区域(11)间隔的多个平行的条状亲水区域(12),所述条状亲水区域(12)垂直于所述免疫印迹卷轴的中心轴,所述条状亲水区域(12)沿其长度方向具有一个或多个结合区(13); 其中,每个所述条状亲水区域(12)分别用于结合不同的待测溶液,从而独立地进行不同的待测溶液中的目标抗体的免疫印迹检测,所述结合区(13)上具有目标抗原,用于与待测溶液中的目标抗体结合。 An immunoblot spool, wherein the spool immunoblotting of outer circumferential surface having a plurality of parallel strip-shaped hydrophilic areas (12) with a hydrophobic region (11) spaced apart, said strip pro water region (12) perpendicular to the central axis of the reel immunoblotting, the hydrophilic region strip (12) having one or more binding regions (13) along its length; wherein each of said strip-like pro water region (12) are different solutions to be tested for binding to independently perform immunoblotting antibody of different solutions to be tested in the detection of the antigen-binding region having a target (13), with a test for binding antibody solution.
  2. 2. 如权利要求1所述的免疫印迹卷轴,其特征在于,包括转轮(17)以及覆盖于所述转轮(17)周向的外表面的免疫印迹薄膜(15);所述转轮(17)用于固定免疫印迹薄膜(15),所述免疫印迹薄膜(15)具有以疏水区域(11)间隔的多个平行的条状亲水区域(12)。 2. immunoblotting reel according to claim 1, characterized in that it comprises a rotating wheel (17) and covering said wheel (17) the outer surface of the film immunoblot circumferential direction (15); said wheel (17) for fixing immunoblotting film (15), the immunoblot films (15) having a plurality of parallel strip-shaped hydrophilic areas (12) with a hydrophobic region (11) spaced apart.
  3. 3. 如权利要求2所述的免疫印迹卷轴,其特征在于,所述免疫印迹薄膜(15)粘贴于所述转轮(17)周向的外表面。 Immunoblotting of the reel as claimed in claim 2, wherein said immunoblotting film (15) attached to an outer surface of the reel (17) in the circumferential direction.
  4. 4. 如权利要求2所述的免疫印迹卷轴,其特征在于,所述免疫印迹薄膜(I5)的底面以及所述转轮(17)周向的外表面具有磁性涂层,所述免疫印迹薄膜(15)通过磁力吸附于所述转轮(17)周向的外表面。 4. The immunoblot reel according to claim 2, characterized in that the outer surface of the film immunoblotting (I5) and a bottom surface of said wheel (17) having a circumferential magnetic coating, the film immunoblotting (15) magnetically adsorbed to said wheel (17) of the outer circumferential surface.
  5. 5. 如权利要求2所述的免疫印迹卷轴,其特征在于,所述转轮(I7)的外周设置有卡扣, 所述卡扣将所述免疫印迹薄膜(15)固定于所述转轮(17)周向的外表面。 5. immunoblotting according to claim 2 reels, wherein the reels (I7) is provided with gripping the outer periphery of said snap immunoblotting the film (15) fixed to said wheel (17) of the outer circumferential surface.
  6. 6. 如权利要求1-5中任意一项所述的免疫印迹卷轴,其特征在于,所述免疫印迹卷轴的直径为2cm〜6cm,宽度为2cm〜6cm。 As claimed in any one of claims 1-5 immunoblot spool, wherein the spool diameter immunoblot 2cm~6cm, width 2cm~6cm.
  7. 7. 如权利要求1 _5中任意一项所述的免疫印迹卷轴,其特征在于,所述条状亲水区域(12)具有多个结合区(13),所述多个结合区(13)包括实验区以及对照区。 7. immunoblotting according to any one of claims 1 _5 spool, characterized in that the strip-shaped hydrophilic zone (12) having a plurality of bonding pads (13), a plurality of bonding pads (13) including experimental area and control area.
  8. 8. 如权利要求7所述的免疫印迹卷轴,其特征在于,所述对照区包括阳性对照区、阴性对照区或结合物对照区。 8. immunoblotting reel according to claim 7, wherein said control zone comprises a positive control area, a negative control region or a control region in combination thereof.
  9. 9. 如权利要求1-5中任意一项所述的免疫印迹卷轴,其特征在于,所述条状亲水区域(12)以及疏水区域(11)的宽度为1mm〜5臟。 As claimed in claim any one of claims 1-5 immunoblotting spool, wherein the width of the strip-shaped hydrophilic zone (12) and a hydrophobic region (11) is 1mm~5 dirty.
CN 201710190186 2017-03-27 2017-03-27 Immunoblotting reel CN107045066A (en)

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Publication number Priority date Publication date Assignee Title
WO2003031979A1 (en) * 2001-10-05 2003-04-17 Surmodics, Inc. Randomly ordered arrays and methods of making and using
CN102445375A (en) * 2010-09-30 2012-05-09 广州阳普医疗科技股份有限公司 Sample adding vessel device
CN103261872A (en) * 2010-10-06 2013-08-21 保科医疗公司 Methods and systems for efficient processing of biological samples
CN104769429A (en) * 2012-06-11 2015-07-08 Abo血型诊断公司 In vitro diagnosis device and uses thereof
CN105527282A (en) * 2014-10-16 2016-04-27 郑兆珉 Fabric-base biochemical detecting device and the fabricating method thereof
CN206892114U (en) * 2017-03-27 2018-01-16 武汉优视科技有限公司 Immunoblotting spool

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003031979A1 (en) * 2001-10-05 2003-04-17 Surmodics, Inc. Randomly ordered arrays and methods of making and using
CN102445375A (en) * 2010-09-30 2012-05-09 广州阳普医疗科技股份有限公司 Sample adding vessel device
CN103261872A (en) * 2010-10-06 2013-08-21 保科医疗公司 Methods and systems for efficient processing of biological samples
CN104769429A (en) * 2012-06-11 2015-07-08 Abo血型诊断公司 In vitro diagnosis device and uses thereof
CN105527282A (en) * 2014-10-16 2016-04-27 郑兆珉 Fabric-base biochemical detecting device and the fabricating method thereof
CN206892114U (en) * 2017-03-27 2018-01-16 武汉优视科技有限公司 Immunoblotting spool

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