CN218474909U - Tandem protein chromatographic column - Google Patents
Tandem protein chromatographic column Download PDFInfo
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- CN218474909U CN218474909U CN202222678438.8U CN202222678438U CN218474909U CN 218474909 U CN218474909 U CN 218474909U CN 202222678438 U CN202222678438 U CN 202222678438U CN 218474909 U CN218474909 U CN 218474909U
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Abstract
The utility model provides a serial-type protein chromatographic column, includes chromatographic column body filling opening, one end with the first chromatographic column of filling opening pipe connection, the first chromatographic column other end and three-way valve connection, three-way valve one end and waste liquid jar are connected, and the three-way valve still is connected with second chromatographic column, and second chromatographic column is connected with the protein collector, the utility model provides a serial-type protein separation column chromatography device has solved in the current protein purification experiment, when using a purification post to obtain purer protein if the protein, and when protein elution component has adverse effect to the stability and the structure of protein itself, can advance the appearance to another kind of packing component's chromatographic column at once after the protein elution on, can realize the replacement and the secondary purification of solution fast, used the three-way valve as the knockout simultaneously, impurity when can effectively avoiding once purifying enters into the second purification post, has avoided the pollution of secondary purification.
Description
Technical Field
The utility model provides a protein purification device in the field of bioengineering, in particular to a tandem type protein chromatographic column.
Background
The protein is a natural product which is composed of 20 amino acids and has a specific three-dimensional structure. The amino acid side chains have different physicochemical properties, and are hydrophilic, hydrophobic, positively charged and negatively charged; having a carboxyl, amino, thiol group; also phenyl-, hydroxyphenyl-, imidazole-and indole-rings; and so on. The primary structure of the protein, i.e., the amino acid sequence of the protein, concatenates these amino acids; the amino acid sequence of a protein determines the three-dimensional structure formed by protein folding and determines the three-dimensional space parameters such as the shape and the size of the protein. The stable three-dimensional structure formed by protein folding determines the charge distribution, the hydrophobic region distribution and the special chemical group distribution on the surface of the protein, determines the property of the protein capable of interacting with other proteins or other substances, and determines the applicable protein separation and purification method. The nature of the different fillers and the nature of the protein determine the nature of the buffer in the protein purification process, but some buffers may adversely affect the stability and structure of the protein, and therefore the purification process takes care of the nature of the buffer to avoid denaturation of the protein as much as possible.
SUMMERY OF THE UTILITY MODEL
The utility model provides a series protein separation column chromatography device. The problem of protein denaturation caused by buffer solution in the prior art is solved; in the present protein purification experiment, if the protein uses a purification column can not obtain purer protein, and when protein elution component has adverse effect to the stability and the structure of protein itself, can advance the appearance to another kind of chromatographic column of packing component at once after the protein elution, can realize the replacement and the secondary purification of solution fast, used the three-way valve as the knockout simultaneously, impurity when can effectively avoiding once purifying enters into the second purification column, avoided the pollution of secondary purification.
In order to achieve the above purpose, the technical scheme of the utility model is realized like this:
the utility model provides a serial-type protein chromatographic column, include chromatographic column body filling opening, one end and the first chromatographic column of filling opening pipe connection, the other end and the three-way valve connection of first chromatographic column, three-way valve one end is connected with the waste liquid jar, the three-way valve still is connected with second chromatographic column, second chromatographic column is connected with albumen collector or detecting instrument.
Further, the first chromatographic column comprises a chromatographic column body, the upper end of the chromatographic column body is connected with the liquid adding port, the lower end of the chromatographic column body is connected with the three-way valve, a filter unit is arranged in the chromatographic column body, a heat exchange layer is arranged on the periphery of the chromatographic column body, and a heat exchange medium inlet for inputting a heat exchange medium and a heat exchange medium outlet for outputting the heat exchange medium are arranged on the heat exchange layer;
the second chromatography column is of the same construction as the first chromatography column.
Furthermore, the filtering unit is a sand core filtering unit.
Furthermore, the first chromatographic column and the second chromatographic column are formed by connecting chromatographic columns consisting of two types of fillers which are the same or different in series, the fillers in the chromatographic columns are set according to protein properties, target proteins can be effectively combined from flowing protein solutions, and the target proteins can be eluted from the fillers by an elution buffer solution.
Further, the first chromatographic column and the second chromatographic column adopt organic glass tube materials.
Further, the protein collector may be replaced with a detection instrument.
The utility model discloses beneficial effect:
the utility model discloses a core is to establish ties two albumen chromatographic column, constitutes a whole, and the initial gross separation of purpose albumen is accomplished to first chromatographic column to discharge impurity through the three-way valve, then change the well liquid flow direction of three-way valve, use second chromatographic column to carry out the secondary purification to the albumen. The two-step purification is changed into one-step purification, and simultaneously, the protein is replaced from the buffer system of the first-step purification as soon as possible.
The utility model provides an advantage of series connection post scheme is no matter what method of adoption is eluted, and elution buffer solution can not appear in the target protein solution of final collection. The final collected target protein is in a buffer solution which is mild to the protein, and the composition of the buffer solution can be optimized according to the requirements of the storage condition of the target protein.
Drawings
Fig. 1 is a schematic view of the main structure of the present invention.
FIG. 2 is the structure diagram of the chromatography column body of the present invention.
In the figure: 1. a first chromatography column; 2. a second chromatography column; 3. a waste liquid tank; 4. a protein collector; 5. a three-way valve; 6. a liquid filling port; 11. a chromatography column body; 12. a filtration unit; 13. a heat exchange layer; 14. a heat exchange medium inlet; 15. and a heat exchange medium outlet.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to the drawings and specific embodiments. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
As shown in figures 1 and 2, the tandem protein chromatographic column comprises a chromatographic column body liquid adding port 6, a first chromatographic column 1 with one end connected with the liquid adding port 6 through a pipeline, the other end of the first chromatographic column 1 is connected with a three-way valve 5, one end of the three-way valve 5 is connected with a waste liquid cylinder 3, the three-way valve 5 is further connected with a second chromatographic column 2, and the second chromatographic column 2 is connected with a protein collector 4 or a detection instrument.
The first chromatographic column 1 comprises a chromatographic column body 11, the upper end of the chromatographic column body 11 is connected with a liquid adding port, the lower end of the chromatographic column body 11 is connected with a three-way valve 5, a filtering unit 12 is arranged in the chromatographic column body 11, a heat exchange layer 13 is arranged on the periphery of the chromatographic column body 11, and a heat exchange medium inlet 14 for inputting a heat exchange medium and a heat exchange medium outlet 15 for outputting the heat exchange medium are arranged on the heat exchange layer 13;
the second chromatography column 2 has the same structure as the first chromatography column 1.
The filter unit 12 is a sand core filter unit.
The first chromatographic column 1 and the second chromatographic column 2 are formed by connecting chromatographic columns consisting of two types of fillers which are the same or different in series.
The first chromatographic column 1 and the second chromatographic column 2 adopt organic glass tube materials.
The protein collector 4 may be replaced with a detection instrument.
The utility model discloses a working process is:
before the experiment is started, the loading buffer, the equilibrium buffer and the elution buffer needed by the first chromatographic column 1 and the second chromatographic column 2 need to be configured respectively. The loading buffer and the balance buffer are used to ensure that the target protein can be combined on the corresponding chromatographic column and the non-combined hybrid protein can be removed.
After the experiment is started, the liquid flow direction of the three-way valve 5 is adjusted to enable the liquid to flow to the second chromatographic column 2, the second chromatographic column 2 is balanced by using a balance buffer solution of the second chromatographic column 2, and the liquid flow direction of the three-way valve 5 is adjusted after the balance is good to enable the liquid to flow to the waste liquid tank 3;
balancing the column by using the balance buffer solution of the first chromatographic column 1, adjusting a three-way valve 5 at the same time, enabling the liquid to flow to a waste liquid tank 3, then dissolving a mixture containing target protein and hetero-protein in a sample loading buffer solution, loading the sample on the first chromatographic column 1, washing by using the sample loading buffer solution and the balance buffer solution until the hetero-protein is cleaned, wherein the target protein is still adsorbed on the first chromatographic column 1, and the solution in the whole column is uniformly the balance buffer solution before elution; changing the elution buffer solution, and continuing chromatography;
simultaneously, adjusting a three-way valve 5 to enable eluent to flow to the second chromatographic column 2, collecting flow-through liquid at the protein collector 4, then replacing the balance buffer solution of the second chromatographic column 2 for washing until the foreign proteins are washed, throwing the target proteins to be adsorbed on the second chromatographic column 2, and uniformly taking the solution in the whole column as the balance buffer solution before elution;
the elution buffer is replaced, chromatography is continued, and the target protein is collected at the protein collector 4. At this time, the eluted target protein is present in the elution buffer of the second chromatography column 2.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
In this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, which may include other elements not expressly listed in addition to those listed. The above description is only for the specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present invention, and all should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (5)
1. The utility model provides a serial-type protein chromatographic column, its characterized in that, including chromatographic column body filling opening (6), one end with first chromatographic column (1) of filling opening (6) pipe connection, the other end and three-way valve (5) of first chromatographic column (1) are connected, three-way valve (5) one end is connected with waste liquid jar (3), three-way valve (5) still are connected with second chromatographic column (2), second chromatographic column (2) are connected with protein collector (4) or detecting instrument.
2. The tandem protein chromatography column of claim 1, wherein the first chromatography column (1) comprises a chromatography column body (11), the upper end of the chromatography column body (11) is connected with the liquid adding port, the lower end of the chromatography column body (11) is connected with the three-way valve (5), a filtering unit (12) is arranged in the chromatography column body (11), a heat exchange layer (13) is arranged on the periphery of the chromatography column body (11), and a heat exchange medium inlet (14) for inputting a heat exchange medium and a heat exchange medium outlet (15) for outputting the heat exchange medium are arranged on the heat exchange layer (13);
the second chromatographic column (2) has the same structure as the first chromatographic column (1).
3. An in-line protein chromatography column according to claim 2, wherein the filtration unit (12) is a sand core filtration unit.
4. The tandem protein chromatography column according to claim 1, wherein the first chromatography column (1) and the second chromatography column (2) are formed by connecting chromatography columns composed of two same or different types of packing in series.
5. The tandem protein chromatography column according to claim 2, wherein the first chromatography column (1) and the second chromatography column (2) are made of organic glass tube material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222678438.8U CN218474909U (en) | 2022-10-12 | 2022-10-12 | Tandem protein chromatographic column |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222678438.8U CN218474909U (en) | 2022-10-12 | 2022-10-12 | Tandem protein chromatographic column |
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| Publication Number | Publication Date |
|---|---|
| CN218474909U true CN218474909U (en) | 2023-02-14 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202222678438.8U Active CN218474909U (en) | 2022-10-12 | 2022-10-12 | Tandem protein chromatographic column |
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| Country | Link |
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| CN (1) | CN218474909U (en) |
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2022
- 2022-10-12 CN CN202222678438.8U patent/CN218474909U/en active Active
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