Patent Application
20180299440
The present invention relates to a high-density western blot array analysis method, and more particularly, to a high-density western blot array analysis method capable of simultaneously checking whether specific proteins are present in a plurality of samples, or simultaneously checking whether a plurality of proteins are present in a single sample.
Western blotting is a research technique for detecting expressed proteins through analysis of a specific sample.
Western blotting is used to detect the presence or the post-translational modification of proteins in cells or tissues.
Roughly, the process of western blotting is performed in the order of SDS-PAGE, protein transfer to a nitrocellulose or PVDF transfer membrane, treatment with an antibody against a target protein, and detection of the target protein through antibody labeling analysis. As samples for western blotting, generally, protein mixtures from cells or tissue are used. Before use in western blotting, a sample is prepared by cell lysis, and then proteins are purified through several times of centrifugation and other separation methods.
To effectively perform the detection of target proteins with a very small amount, it is necessary to widely spread proteins in the sample, and to this end, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is frequently used. SDS binds to a protein in a sample such that the proteins lose secondary, tertiary and quaternary structures and become linear, and a high density of negative charges are imparted to the protein to be attracted towards the positive electrode in electrophoresis. That is, SDS-PAGE as described above is for sorting proteins denatured with SDS by size on a polyacrylamide gel (PAG) through electrophoresis.
Since the proteins separated by SDS-PAGE are buried inside of the gel, observation of direct binding between a probe and the protein is difficult.
Therefore, the protein is transferred onto a thin transfer membrane to facilitate binding of a probe to the protein.
Among a variety of such transfer methods, the most widely used method is an electrophoretic transfer method.
This method uses a very similar principle to electrophoresis, and two methods have been conventionally used.
One is a semi-dry transfer method for transferring proteins by placing a gel horizontally, and the other is a wet transfer method for transferring proteins while a gel is vertically placed in a tank filled with buffer.
Here, in the transfer method, after the electrophoresis, the gel is carefully separated, and placed onto the transfer membrane.
Then, filter papers and pads are placed at both sides of a gel and transfer membrane to sufficiently absorb a buffer and to protect the gel, and a negative electrode and a positive electrode are placed at both sides thereof.
Here, since a protein is attracted to the positive electrode, the transfer membrane is placed at the positive electrode side.
When a voltage is applied, proteins gradually begin to be transferred from the gel to the membrane, resulting in attachment onto the transfer membrane.
When the proteins are transferred onto the transfer membrane, a blocking process is performed to prevent attachment of other proteins to the transfer membrane.
This blocking process allows proteins which do not react with previously known probes to be attached to all sites except those to which the sample proteins are attached.
The probe antibody is produced from lymphocytes, and has a property of specifically binding only to a target protein.
That is, the probe needs to be previously prepared for an experiment, and different types of probes should be used depending on target proteins.
Therefore, conventional western blotting had problems of excessive consumption of required antibodies and demand for an excessive amount of a sample.
As a result, there was a limit to the number of samples used at once.
Moreover, a conventional western blotting system requires much time consumption for electrophoresis, and a long electrophoretic distance for migration of a sample.
Meanwhile, in U.S. Unexamined Patent Application Publication No. 2011-0028339 as prior art, a highly-integrated western blotting technique is disclosed.
However, U.S. Unexamined Patent Application Publication No. 2011-0028339 has many problems, for example, a demand for high-priced equipment, and difficulty in freely configuring test formats, particularly, limitation of the number and types of samples that can be tested at one time.
(Patent Document 1) U.S. Pat. No. 8,940,232 B2
(Patent Document 2) U.S. Pat. No. 7,670,833 B2
(Patent Document 3) US2011-0028339 A1
One object of the present invention is to overcome the limitation of a conventional western blotting, and to implement a highly-integrated array technique for performing multiple western blotting simultaneously.
In addition, another object of the present invention is to provide gel formers, a sample injection unit and antibody incubator chambers, which are necessary for implementing the above-described technique.
To achieve the above-mentioned objects, one aspect of the present invention provides a high-density western blot array analysis method, which includes: preparing a gel in which a plurality of sample injection holes are arranged in one or more rows; injecting a sample into each sample injection hole in the gel; electrophoresing the sample-injected gel in a direction perpendicular to the row of the sample injection holes; transferring the sample in the electrophoresed gel to a membrane; putting the entire sample-transferred membrane into one area formed in an antibody incubation chamber and treating the membrane with an antibody; and analyzing the antibody-treated membrane.
To achieve the above-mentioned objects, another aspect of the present invention provides a high-density western blot array analysis method, which includes: preparing a gel in which one or more sample injection slots each having a predetermined width are arranged in a longitudinal direction of the width so as to form one or more rows; injecting a sample into the sample injection slots in the gel; electrophoresing the sample-injected gel in a direction perpendicular to the array of the sample injection slots; transferring the samples in the electrophoresed gel to a membrane; placing the sample-transferred membrane, in an antibody incubation chamber in which a plurality of antibody injection holes and a plurality of microchannels are formed to fit the width of the sample injection slots, and then treating the sample on the membrane with antibodies in microchannels by injecting different types of antibodies into the respective antibody injection holes; and analyzing the antibodies-treated membrane.
According to the present invention as described above, the following effects are exhibited:
First, it is possible to perform high-density western blotting, resulting in the decrease in the volume of a sample;
Second, analysis can be performed even with a very small amount of an antibody;
Third, the number of samples which can be used in one experiment is greatly increased;
Fourth, the time required for the experiment is greatly reduced;
Fifth, an electrophoresis distance is shortened and therefore test instruments become compact;
Seventh, there are no need of high-priced equipment and no limitation of test formats; and
Eighth, since there is a sealing member between an upper frame and a lower frame, an antibody incubation chamber is sealed.
Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.
In this process, the thicknesses of the lines and the sizes of the components shown in the drawings may be exaggerated for clarity and convenience of explanation. In addition, the following terms are defined in consideration of the functions of the present invention, and this may vary according to the intention or custom of the user or operator. Therefore, the definition of these terms should be based on the contents throughout the specification.
One aspect of the present invention provides a high-density western blot array analysis method (first analysis method) for checking whether a protein binding to a single antibody is present in a plurality of samples. The first analysis method is multi sample analysis, for example, may be applied to the analysis using a drug library, a biodrug library, an RNAi library or an antisense RNA library, may be used as an alternative to high throughput screening (HTS) used in in vitro screening, and may be used in screening for various target molecules.
Referring to
Referring to
Each of the plurality of samples injection holes 110 has a size that allows several microliters, preferably, 0.1 to 5 μl, and more preferably 0.5 to 2 μl of a sample to be injected.
20 or more, preferably 50 or more, and more preferably 100 or more of the sample injection holes 110 may be arranged at an interval of several mm, preferably 0.5 to 5 mm, and more preferably 1 to 3 mm to form one row.
In addition, there are a plurality of rows consisting of the plurality of samples injection holes 110. When there are a plurality of rows, the plurality of rows are formed to have an interval larger than the distance at which the sample injected into the sample injection hole is electrophoresed, for example, at an interval of several cm, preferably, 0.5 to 5 cm, and more preferably, 1 to 3 cm.
Specifically, as shown in
In an exemplary embodiment of the present invention, 101 sample injection holes arranged at an interval of 1 mm form one row, and a gel having a size of 12 cm×6 cm in which four rows formed as described above are arranged at an interval of 1.2 cm was prepared.
As shown in
Referring to
The main body 310 includes a first plate 311 formed of a flat plate-shaped member and formed in a rectangular shape, and a second plate 313 formed at an edge of the top surface of the first plate at a predetermined height. The second plate 313 may be formed in a “⊏” shape, and a concave space formed by the flat plate-shaped first plate 311 and the “⊏” shaped second plate 313 becomes a gel container 330 into which gel is injected. As a result, the first plate 311 becomes a bottom surface of the gel container 330, and the second plate 313 becomes a side surface of the gel container 330. Additionally, a sealing member S may be disposed to prevent leakage of the gel out of the gel container 330, and a plurality of clamping apparatuses 315 may be disposed at a predetermined interval on a top surface of the second plate 313.
The cover unit 350 includes a frame 351 and a molding plate 353 including a plurality of molding projections 355. The molding projections 355 are formed so as to protrude from one surface of the molding plate 353, and sample injection holes 110 having a size to allow a volume of several microliters of injected sample are formed by inserting at least some projections into the gel container 330. The molding projections 355 are formed in one or more rows at an interval of several mm, and when there are a plurality of rows consisting of the molding projections 355, the rows may be arranged at an interval of several cm. The molding plate 353 may be integrated with the frame 351, or is more preferably detachable, which is for being replaced according to the pattern, interval, size or number of the plurality of molding projections 355.
The cover unit 350 may be installed to the main body 310 so as to be movable between a first position for opening the gel container 330 and a second position for closing the gel container 330. More preferably, the cover unit 350 may be pivotally coupled to one side of the first plate 311. When the cover unit 350 is disposed at the second position for closing the gel container 330, at least some of the molding projections 355 of the molding plate 353 are inserted into the gel injected into the gel container 330 to make sample injection holes.
While the gel container 330 is sealed by the cover unit 350 at the second position, the cover unit 350 is fixed to the main body 310 by the clamping apparatuses 315. As the sealed state of the gel container 330 by the cover unit 350 using the clamping apparatuses 315 is maintained for a predetermined time, the gel contained in the gel container 330 may be hardened while the sample injection holes 110 corresponding to the shape of the molding projections 355 are formed.
The first gel former 300 may further include a supporting member 370. The supporting member 370 includes a first supporting member 371 and a second supporting member 373, in which the first supporting member 371 supports the bottom of one side of the main body 310, and the second supporting member 373 supports the bottom of the other side of the main body 310. More preferably, the first supporting member 371 is fixed to the lower part of the first plate 311 at a connection side at which the cover unit 350 is pivotably coupled to the main body 310, and the second supporting member 373 is spaced apart from the first supporting member 371 to be fixed to the lower part of the first plate 311 in a longitudinal direction. More preferably, the first supporting member 371 is extended to a direction of the lower part of the first plate 311, and the second supporting member 373 is extended to a direction of the lower and upper parts of the first plate 311. The part that is extended to the lower direction is a second supporting member 373, and the part that is extended to the upper direction is a third supporting member 375. The second supporting member 373 and the third supporting member 375 are formed in a flat plate shape, and support on the ground such that the main body 310 can be set up in the vertical direction. In other words, while the cover unit 350 is fixed with the clamping apparatuses 315 at the second position for closing the gel container 330, the main body 310 may be set up in the vertical direction to the ground by the second supporting member 373 and the third supporting member 375. In addition, since the first supporting member 371 is formed higher than the second supporting member 373, when the main body 310 is disposed on the ground in a horizontal direction, the first plate 311 may be inclined. Meanwhile, the second plate 313 may be formed in a “⊏” shape, the ends of the “⊏” shape face parts at which the cover unit 350 is connected to the first plate 311, respectively. In other words, the second plate 313 is formed along both lateral edges in a longitudinal direction and one widthwise edge on the top surface of the first plate 311, thereby forming the “⊏” shape.
A mesh-type support film for fixing the gel is inserted into the gel container 330, the cover unit 350 is moved pivotally to be placed at the second position for closing the gel container 330, and then fixed to the second position using the clamping apparatuses 315. From the second position at which the gel container 330 is closed by the cover unit 350 while the supporting member 370 is placed on the ground, a liquid gel is injected into the space between the gel container 330 and the molding plate 353. Since the first supporting member 371 is formed longer than the second supporting member 373, the gel flows on the surface of the first plate 311 to fill the space between the gel container 330 and the molding plate 353. The second supporting member 373 and the third supporting member 375 may be put on the ground, such that the main body 310 of the gel former may be set up on the ground in the vertical direction. After a predetermined time, when the gel in the gel container 330 is hardened, the clamping apparatuses 315 are unfastened to allow the cover unit 350 to be moved back to the first position and to allow the gel container 330 to be open, and then the gel 100 in which the sample injection holes 110 are molded to correspond to the shape of the molding projections 355 is separated from the main body 310, thereby obtaining the gel 100 used in the first analysis method of the present invention as shown in
Subsequently, a sample to be analyzed is injected into the sample injection holes of the gel prepared as described above (S2). A different sample may be injected into each of the plurality of samples injection holes, and the volume of each sample to be injected may be several microliters, preferably, 0.1 to 5 μl, and more preferably 0.5 to 2 μl. In one exemplary embodiment of the present invention, 1 μl each of different samples was injected into each of 384 grooves among the 404 sample injection holes formed in the gel, and 1 μl of a size marker was injected into each of the remaining 20 sample injection holes.
When there are a plurality of sample injection holes formed in the gel as described above, a sample injection unit 400 as shown in
Referring to
The rod unit 410 is formed to facilitate grasping of the sample injection unit 400. The pins 430 are inserted into the sample injection holes of a gel to inject a sample into each of the sample injection holes. Here, since the gel prepared by the first gel former is very vulnerable, when the sample is injected into the sample injection holes formed in the gel, in some cases, the pins 430 may break the gel G. Therefore, as the plurality of pins 430 are formed to slide through the second plate 422 even when applying a predetermined force when the sample is injected into the sample injection holes formed in the gel the gel G is not damaged. In other words, since the first plate 421 is spaced apart from the second plate 422, the plurality of pins 430 may slide in a gap 425 between the first and second plates, and thus the gel G may not be destroyed.
After the sample is injected into the sample injection holes of the gel as described above, the sample-injected gel is electrophoresed (S3). The electrophoresis is a process of separating proteins of the samples injected into the sample injection holes, and is performed using a horizontal electrophoresis device (not shown). The electrophoresis is performed in a direction perpendicular to a row of the sample injection holes for several minutes, preferably, 10 to 60 minutes, and more preferably 20 to 40 minutes.
After the electrophoresis is completed as described above, the sample separated in the gel is transferred onto a membrane (S4). The transfer process of the sample may be performed according to a method conventionally used in the art.
After the sample is transferred onto the membrane as described above, the membrane is treated with an antibody (S5).
In the first analysis method, the sample-transferred membrane may be treated with one type of antibody. That is, the same antibody may be treated against a plurality of different samples, and therefore, it is possible to simultaneously confirm the absence or presence of a specific protein in the plurality of different samples.
In one embodiment of the present invention, after a total of 384 samples are treated with the same antibody, the absence or presence of specific proteins in the 384 samples was confirmed.
In the first analysis method of the present invention, as shown in
Referring to
In the upper frame 510, the first through hole 512 through which an antibody is provided and the second through hole 511 through which a waste solution remaining after the reaction is extracted are formed, and when the upper frame 510 is coupled to the lower frame 520, the first and second through holes 512 and 511 in the upper frame 510 communicate with the space 521 in the lower frame 520.
When the sample-transferred membrane is placed in the space 521 communicating with the first and second through holes as described above, the sample transferred onto the membrane reacts with an antibody by injecting a reaction material including the antibody into the first through hole 512, and after the above-mentioned reaction is completed, the waste solution is extracted through the second through hole 511. Afterward, the first and second through holes 512 and 511 may also be used in blocking or washing of the membrane through the injection and discharging of a blocking buffer or washing buffer.
The space 521 has a flat part 550 adjacent to the first through hole 512 at one side and an angled part 540 adjacent to the second through hole 511 at the other side. The angled part 540 is formed by meeting a first side 541 with a second side 542 at a predetermined angle, such that a waste solution or washing buffer solution is gathered at the angled part 540 to be entirely extracted to the outside.
Meanwhile, in the space 521, partitions 525 may be further included as shown in
After the antibody treatment for the membrane is completed, the membrane is analyzed (S6). The analysis process of the membrane may go through exposure and development, or may be performed according to a method conventionally used in the art such as reading data using a fluorescence scanner.
Another aspect of the present invention provides a high-density western blot array analysis method (second analysis method) for simultaneously checking the absence or presence of proteins binding to a plurality of antibodies in a single sample. The second analysis method may be a multi-antibody screening system, and may be actively applied to protein analysis using an antibody library, for example, protein PTM analysis, and therefore several tens to several hundreds of types of antibodies may be simultaneously analyzed as markers.
Referring to
Referring to
Each of the one or more sample injection slots 210 may have a size such that several to several tens of microliters, preferably 0.1 to 50 and more preferably 1 to 20 μl of a sample can be injected.
The sample injection slots 210 have a predetermined width, for example, several tens of mm, preferably 10 to 80 mm, and more preferably 30 to 60 mm.
There may be one sample injection slot 210 formed in a row, or two or more sample injection slots 210 formed in one row. As described above, when a plurality of sample injection slots 210 are formed in one row, the sample injection slots 210 may be arranged at an interval of several mm, preferably 0.5 to 5 mm, and more preferably 1 to 3 mm, and a marker injection hole 215 which can inject a marker may be additionally formed between the sample injection slots 210.
In addition, there may be a plurality of rows consisting of the one or more sample injection slots 210. When there are a plurality of rows, the plurality of rows may be formed to have a wider gap than the distance of electrophoresing the sample injected into each sample injection slot 210, and for example, may be arranged at an interval of several cm, preferably 0.5 to 5 cm, and more preferably 1 to 3 cm.
In one exemplary embodiment of the present invention, four sample injection slots having a width of 45 mm were formed in one row, and a gel having a size of 12 cm×6 cm in which such four rows are arranged at an interval of 1.2 cm was prepared.
Such a gel 200 used in the second analysis method of the present invention may be prepared using a second gel former 350 as shown in
Referring to
The molding projections 355′ are formed in a straight line with a width of several tens of millimeters, preferably 10 to 80 mm, and more preferably 30 to 60 mm, such that one or more straight lines are formed in one or more rows at an interval of several mm. When there are a plurality of rows of the molding projections 355′, the rows may be arranged at an interval of several cm. The molding plate 355′ may be integrated with a frame 351′ or is preferably detachable from the frame 351′ so as to be replaced depending on the pattern, gap, size or number of the plurality of molding projections 355′.
In the case of the second gel former 350, a mesh-type support film for fixing the gel is inserted into the gel container 330, the cover unit 350′ is moved pivotally to be placed at the second position for closing the gel container 330, and then fixed to the second position using the clamping apparatuses 315. From the second position at which the gel container 330 is closed by the cover unit 350′ while the supporting member 370 is placed on the floor, a liquid gel is injected into the space between the container 330 and the molding plate 353′. Since the first supporting member 371 is formed longer than the second supporting member 373, the gel flows on the surface of the first plate 311 to fill the space between the gel container 330 and the molding plate 353′. The second supporting member 373 and the third supporting member 375 may be put on the floor, such that the main body 310 of the gel former may be set up on the floor in the vertical direction. After a predetermined time, when the gel in the gel container 330 is hardened, the clamping apparatuses 315 are unfastened to allow the cover unit 350′ to be moved back to the first position and to allow the gel container 330 to be open, and then the gel 200 in which the sample injection slots 210 and marker injection holes 215 are molded to correspond to the shape of the molding projections 355′ is separated from the main body 310, thereby obtaining the gel 200 used in the second analysis method of the present invention.
Subsequently, a sample to be analyzed is injected into the sample injection slots of the gel prepared as described above (S2). A different sample may be injected into each of the plurality of samples injection slots, and the volume of each sample to be injected may be several to several tens of microliters, preferably, 0.1 to 50 μl, and more preferably 1 to 20 μl, and when there are a plurality of sample injection slots, different samples may be injected into the sample injection slots, respectively. In one exemplary embodiment of the present invention, 20 μl of a different sample was injected into each of 16 sample injection slots formed in the gel.
In this second analysis method, since the number of the sample injection slots is not high, unlike the first example, the sample injection unit may not be used, and a common micropipette may be used.
After the sample is injected into the sample injection slots of the gel as described above, the sample-injected gel is electrophoresed (S3). The electrophoresis is a process of separating proteins of a sample injected into each sample injection slot, and is performed using a horizontal electrophoresis device (not shown). The electrophoresis is performed in a direction perpendicular to a row of the sample injection slots for several minutes, preferably, 10 to 60 minutes, and more preferably 20 to 40 minutes.
After the electrophoresis is completed as described above, the sample separated in the gel is transferred onto a membrane (S4). The transfer process of the sample may be performed according to a method conventionally used in the art.
After the sample is transferred onto the membrane as described above, the membrane is treated with an antibody (S5).
In the second analysis method, the sample isolated from one sample injection slot may be treated with a plurality of different antibodies. That is, as one sample is treated with a plurality of antibodies, the absence or presence of various proteins in one sample may be simultaneously checked.
However, when the sample injection slots are plural, and different samples are injected into the plurality of sample injection slots, respectively, the absence or presence of various proteins in different samples may be simultaneously checked.
Here, the plurality of antibodies may be treated on one sample at a volume of several microliters, preferably, 0.1 to 5 μl, and more preferably, 0.5 to 2 μl.
In one embodiment of the present invention, after one sample is treated with 20 different antibodies, the absence or presence of 20 specific proteins in each of the total of 16 samples was simultaneously checked.
In the second analysis method of the present invention, as shown in
Referring to
Referring to
The mounting part 621 on which the membrane is placed is formed in the lower frame 620, and the lower frame 620 is connected with the upper frame 610 such that the lower surface of the upper frame 610 is in close contact with the upper surface of the membrane placed on the mounting part 621. Particularly, if the membrane placed in the mounting part 621 has a predetermined thickness, the mounting part 621 may have a separate space, or if the membrane is very thin and thus substantially needs no separate space, the mounting part 621 may be a certain part present in the lower frame 620, rather than a separately formed space.
In the second antibody incubation chamber 600, fasteners 630 may be additionally formed to firmly fix the coupling between the upper frame 610 and the lower frame 620.
When the sample-transferred membrane is placed in the space of the mounting part 621 of the lower frame 620 and is in close contact with the lower surface of the upper frame 610 (when there is a sealing member 670, being in close contact with the sealing member 670), the sample transferred onto the membrane reacts with an antibody by injecting a reaction material including different types of antibodies into the plurality of antibody injection holes 611, respectively.
The antibody injection holes 651 and the waste solution discharge holes 653 are used to inject and discharge a washing solution for washing the membrane, as well as the antibodies and a waste solution thereof. The injection of the washing solution into the antibody injection holes 651 and the suction of the waste solution into the waste solution discharge holes 653 are performed by a washing unit 700 as shown in
When an antibody is treated, and a reaction between the treated antibody and the sample in the membrane is completed, the injection tubes 751 and the discharge tubes 753 of the washing unit 700 are inserted into the antibody injection holes 651 and the waste solution discharge holes 653 formed in the upper frame 610, and then, first, a waste solution of the antibody present in the microchannels 655 is sucked through the discharge tubes 753 inserted into the waste solution discharge holes 653 using a vacuum pump (After being sucked as described above, the waste solution is discharged to a waste solution container). Afterward, when the washing solution stored in the washing solution tank is injected into the antibody injection holes 651 through the injection tubes 751, respectively, the washing solution injected as described above flows to the third microchannel 655c through the first microchannel 655a and the second microchannel 655b so as to remove non-binding antibodies present on the membrane. After the washing is completed, a waste solution of the washing solution present in the microchannels 655 is sucked through the discharge tubes 753 inserted into the waste solution discharge holes 653 using a vacuum pump (After being sucked as described above, the waste solution is discharged to a waste solution container).
Meanwhile, in the upper frame 610 of the second antibody incubation chamber 600, as shown in
When the fixing groove 690 are formed in the upper frame 610 of the second antibody incubation chamber 600, the washing unit 700 may include a fixing end 750 which can fix the washing unit 700 by being inserted into the fixing grooves 690, and the injection tubes 751 and the discharge tubes 753 may be included under or may be disposed in the fixing end 750. When the washing unit 700 is inserted into the first and second fixing grooves 691 and 693 and fixed, the numbers of the injection tubes 751 and the discharge tubes 753 of the washing unit 700 may be smaller than those of the antibody injection holes 651 and the waste solution discharge holes 653, and moreover, the injection tubes 751 and the discharge tubes 753 may not be directly inserted into the antibody injection holes 651 and the waste solution discharge holes 653, respectively. In this case, when the washing unit 700 is fixed to the first and second fixing grooves 691 and 693, the injection tubes 751 and the discharge tubes 753 are disposed a predetermined distance apart from the antibody injection holes 651 and the waste solution discharge holes 653 in the first fixing groove 691 and the second fixing groove 693, respectively. The washing solution injected into the injection tubes 751 is injected into the antibody injection holes 651 present in the entire first fixing grooves 691 through the space between the injection tubes 751 and the antibody injection holes 651, and as a sucking power is applied to the entire second fixing groove 693 by a vacuum pump, a waste solution of the washing solution is discharged to the space between the discharge tubes 753 and the waste solution discharge holes 653 present in the entire second fixing groove 693.
After the antibody treatment for the membrane is completed, the membrane is analyzed (S6). The analysis process of the membrane may go through exposure and development, or may be performed according to a method conventionally used in the art such as reading data using a fluorescence scanner.
The present invention has been explained with reference to exemplary embodiments, but it will be understood by those of ordinary skill in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the present invention as described in the accompanying claims.
100: gel 110: sample injection hole
200: gel 210: sample injection slot 215: marker injection hole
300: first gel former 300′: second gel former 310: main body
330: gel container 350, 350′: cover unit 370: supporting member
311: first plate 313: second plate 315: clamping apparatus
351: frame 353: molding plate 355: molding projection
351′: frame 353′: molding plate 355′: molding projection
371: first supporting member 373: second supporting member 375: third supporting member
400: injection unit 410: rod unit
421: first plate 422: second plate 430: pin
500: first antibody incubation chamber 525: partition
510: first upper frame 512, 511: first through hole and second through hole
520: first lower frame 521: chamber space
530: first fastener 540: angled part
541: first side 542: second side 550: flat part
600: second antibody incubation chamber
610: upper frame 620: lower frame 621: mounting part
650: hole pair 651: antibody injection hole 653: waste solution discharge hole
655: microchannel 655a, 655b, 655c: first, second and third microchannels
630: second fastener 670: sealing member
690: fixing groove 691: first fixing groove 693: second fixing groove
700: washing unit 710: frame 750: fixing end
751: injection tube 753: discharge tube
S: sealing member
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2015-0181680 | Dec 2015 | KR | national |
| 10-2016-0172742 | Dec 2016 | KR | national |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/KR2016/014836 | Dec 2016 | US |
| Child | 16010031 | US |
