CN207401184U - endotoxin removal kit - Google Patents
endotoxin removal kit Download PDFInfo
- Publication number
- CN207401184U CN207401184U CN201721441894.3U CN201721441894U CN207401184U CN 207401184 U CN207401184 U CN 207401184U CN 201721441894 U CN201721441894 U CN 201721441894U CN 207401184 U CN207401184 U CN 207401184U
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- China
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- endotoxin
- apyrogeneity
- chromatographic column
- affine resin
- removal kit
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- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The utility model discloses endotoxin removal kit, including a box body；One chromatographic column, the upper end port are plugged with a lid, and protection liquid and affine resin have from top to bottom been sequentially filled in the chromatographic column, and the affine resin is equipped with Endotoxin adsorption aglucon；One regeneration buffer, for activating the affine resin；One level pad, for balancing the affine resin；One flow speed controller is connected on the liquid outlet of the chromatographic column；And several apyrogeneity receiving bottles and several apyrogeneity pipette tips.The kit by providing apyrogeneity buffer solution, apyrogeneity receiving flask, apyrogeneity pipette tips and chromatographic column equipped with affine resin, can high biological safety removal biological products endotoxin, range of applicability is wide, does not influence the activity of most biological samples；Without the i.e. adjustable flow velocity of constant flow pump；It is repeatable to operate with removal endotoxin.
Description
Technical field
Field is isolated and purified the utility model is related to biological products, more particularly to a kind of endotoxin removal kit.
Background technology
Bacterial endotoxin（Lipopolysaccharides, LPS）It is the main component of gram-negative bacterial cell wall.People are blue in research leather
During family name's negative bacteraemia and endotoxemia, it is found that bacterial endotoxin plays crucial effect wherein, endotoxin makes
Body's immunity is badly damaged, and further development may cause septic shock, disseminated intravascular coagulation, acute respiration
Distress Syndrome, systemic inflammatory response syndrome or mortality multiple organ failure.Therefore, endotoxic removal is for the mankind
The injection-type drug of disease treatment, biological products etc. are all necessary, are cloned particularly with genomic medicine, protein drug, monoclonal antibody
Just seem extremely important for the downstream purification processes of biological products such as drug, biovaccine, small-molecule chemical drug.
In the prior art, have efficient endotoxin removal filter, can induced by endotoxin effectively removed, but there is no
The endotoxin removal kit of comprehensive high biological safety.
Utility model content
To solve the above-mentioned problems, the utility model is intended to provide endotoxin removal kit, can high biological safety
To genomic medicine, protein drug, monoclonal antibody drug, biovaccine, small-molecule chemical drug, cell culture, biological medicine
The endotoxic removal of the biological products such as production.
To achieve the above object, the utility model is the technical scheme adopted is that endotoxin removal kit, including：
One box body；
One chromatographic column, the upper end port are plugged with a lid, be from top to bottom sequentially filled in the chromatographic column protection liquid and
Affine resin, the affine resin are equipped with Endotoxin adsorption aglucon；
One regeneration buffer, for activating the affine resin；
One level pad, for balancing the affine resin；
One flow speed controller is connected on the liquid outlet of the chromatographic column；
And several apyrogeneity receiving bottles and several apyrogeneity pipette tips.
The kit of the utility model is by providing apyrogeneity buffer solution, apyrogeneity receiving flask, apyrogeneity pipette tips and dress
Have the chromatographic column of endotoxin Specific adsorption aglucon, can high biological safety removal biological products endotoxin, range of applicability
Extensively, the activity of most biological samples is not influenced；Without the i.e. adjustable flow velocity of constant flow pump；It is repeatable to operate with removal endogenous toxic material
Element.
Preferably, the protection liquid is 20% ethyl alcohol.When chromatographic column to be preserved, 20% ethyl alcohol is filled in affine resin
Top can protect the Endotoxin adsorption aglucon in affine resin.
Preferably, the affine resin is 4% crosslinked Ago-Gel.
Preferably, the grain size of described 4% crosslinked Ago-Gel is 80um-100um.
Preferably, the pipette tips that the apyrogeneity pipette tips wrap 1ml for 6.
Preferably, the Endotoxin adsorption aglucon is polymyxin B or poly-D-lysine.The Endotoxin adsorption of the program is matched somebody with somebody
Base can remove more than 95% endotoxin, and endotoxin content after purification is less than 0.1EU/ml.
The advantageous effect of endotoxin removal kit provided by the utility model is：
Compared with prior art, this kit is easy to use, by providing apyrogeneity buffer solution, apyrogeneity receiving flask, nothing
Pyrogen pipette tips and the chromatographic column equipped with endotoxin Specific adsorption aglucon, can high biological safety removal biological products endogenous toxic material
Element, range of applicability is wide, does not influence the activity of most biological samples；Without the i.e. adjustable flow velocity of constant flow pump；Repeatable behaviour
Make using removal endotoxin.More than 95% endotoxin is can remove, endotoxin content after purification is less than 0.1EU/ml.
Description of the drawings
Fig. 1 is the reagent cartridge configuration schematic diagram of the utility model.
Specific embodiment
The utility model is further illustrated in conjunction with the drawings and specific embodiments.
As shown in Figure 1, endotoxin removal kit, including a box body 1, includes 10, one bottles of a chromatographic column in box body 1
20% second of 30, flow speed controllers of level pad, 40, one bottles of 50ml of 20, one bottles of 125ml of regeneration buffer of 125ml
The apyrogeneity pipette tips 70 of alcohol 50, two bag apyrogeneity receiving bottles 60 and 6 bag 1ml.Often there is the nothing of 3 in bag apyrogeneity receiving bottle 60
Pyrogen receiving bottle 60；Often there are the apyrogeneity pipette tips 70 of 4 in bag apyrogeneity pipette tips 70.
The upper end port of chromatographic column 10 is plugged with a lid 11, and two sections are divided into chromatographic column 10, and wherein epimere is loading
Section 12, hypomere are packing section 13.Loading section 12 is during use, for adding buffer solution and sample；In the shape not used
Under state, preserved for filling 20% ethyl alcohol.Filled with affine resin carrier in packing section 13, affine resin is in the present embodiment
It is middle to use 4% crosslinked Ago-Gel；And 4% crosslinked Ago-Gel grain size be 80um-100um.4% crosslinked agar
It is crosslinked with that endotoxic Endotoxin adsorption aglucon can be adsorbed in sugared gel, Endotoxin adsorption aglucon is using more in the utility model
Colistin B or poly-D-lysine.Certainly other endotoxic substances of absorption in existing can also be used.Polymyxin B or poly
Lysine has higher adsorption capacity, can remove more than 95% endotoxin, and endotoxin content after purification is less than 0.1EU/ml.
Then flow speed controller 40 includes fluid tube 41 and regulating valve 42, and the nozzle of fluid tube 41 is connected with the liquid outlet of chromatographic column 10；
Regulating valve 42 is mounted on the middle part of fluid tube 41, for controlling the flow velocity of the liquid of chromatographic column 10.
To more fully understand the utility model, the utility model is made further with reference to the application method of the utility model
It illustrates.
1st, sample treatment：Sample centrifuges before loading or with 0.22um or 0.45um membrane filtration, to reduce impurity, improves
Protein purification efficiency and prevent block chromatographic column 10.Then sample adjusts PH with the sodium hydroxide of 0.1M and the hydrochloric acid of 0.1M
Value makes sample P H controls in 7-8.Finally, the ionic strength of sample is controlled using the sodium chloride of 0.1M-0.5M, it is non-reduces sample
Specific adsorption.
2nd, affine resin is activated：Chromatographic column 10 is placed in iron stand vertically to fix, removes lid 11, opens flow speed controller
40,20% ethyl alcohol in the loading section 12 of chromatographic column 10 is made to drain off under the effect of gravity；Then 5ml is added in again in loading section 12
Raw buffer solution adjusts flow speed controller 40, keeps flow velocity in 0.25ml/min；Buffer solution to be regenerated drains off, and adds 5ml regeneration
Buffer solution repeats operation twice, it is ensured that in chromatographic column 10 apyrogeneity is kept to exist.
3rd, affine resin is balanced：Activation finishes, and adds in 6ml level pads, uniformly adds along the column tube inner wall of chromatographic column 10
Enter the inner wall wherein, ensured in cleaning column tube, adjust flow speed controller 40, keeping flow velocity, drain off equalizing and buffering in 0.5ml/min
Liquid, then be repeated twice by this operation.
4th, endotoxin removal：Take 1.5ml samples be added to balance after chromatographic column 10 in, adjust flow velocity in 0.25ml/min,
Do not receive efflux.When sample flow is complete, continuing to add sample, sample can be filled it up in the column tube of chromatographic column 10, while using no heat
Former receiving bottle 60 collects efflux, is the sample of depyrogenation.In order to reduce the loss of sample, after sample flow is complete, then past chromatography
Addition 1.5ml level pads in column 10, and merge with efflux before.Detect sample concentration and level of endotoxin.
5th, reuse：If outflow sample level of endotoxin fails to reach desired value, it is necessary to by chromatographic column 10 according to step
Rapid 2 and step 3 regenerate, balance again, then loading purify.
6th, preservation condition：If chromatographic column 10 needs to preserve after being finished, first with 5ml equilibration buffers, ready to balance delays
Fliud flushing drains off, and adds in 6ml20% ethyl alcohol in loading section 12, and in 4 DEG C of preservations.
It these are only the preferred embodiment of the utility model, not thereby limit its scope of the claims, for the skill of this field
Other various corresponding changes and deformation are made in technical solution that can be as described above for art personnel and design, and
All these change and deformation should all belong within the protection domain of the utility model claims.
Claims (6)
1. endotoxin removal kit, it is characterised in that including：
One box body；
One chromatographic column, the upper end port are plugged with a lid, and protection liquid and affine has from top to bottom been sequentially filled in the chromatographic column
Resin, the affine resin are equipped with Endotoxin adsorption aglucon；
One regeneration buffer, for activating the affine resin；
One level pad, for balancing the affine resin；
One flow speed controller is connected on the liquid outlet of the chromatographic column；
And several apyrogeneity receiving bottles and several apyrogeneity pipette tips.
2. endotoxin removal kit as described in claim 1, it is characterised in that：The protection liquid is 20% ethyl alcohol.
3. endotoxin removal kit as described in claim 1, it is characterised in that：The affine resin is 4% crosslinked agar
Sugared gel.
4. endotoxin removal kit as claimed in claim 3, it is characterised in that：The grain of the 4% crosslinked Ago-Gel
Footpath is 80um-100um.
5. endotoxin removal kit as described in claim 1, it is characterised in that：The rifle that the apyrogeneity pipette tips wrap 1ml for 6
Head.
6. endotoxin removal kit as described in claim 1, it is characterised in that：The Endotoxin adsorption aglucon is more Acarasiales
Plain B or poly-D-lysine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721441894.3U CN207401184U (en) | 2017-11-02 | 2017-11-02 | endotoxin removal kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721441894.3U CN207401184U (en) | 2017-11-02 | 2017-11-02 | endotoxin removal kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN207401184U true CN207401184U (en) | 2018-05-25 |
Family
ID=62317537
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201721441894.3U Active CN207401184U (en) | 2017-11-02 | 2017-11-02 | endotoxin removal kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN207401184U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115029344A (en) * | 2022-06-16 | 2022-09-09 | 通用生物(安徽)股份有限公司 | Preparation method of high-standard endotoxin-free plasmid |
-
2017
- 2017-11-02 CN CN201721441894.3U patent/CN207401184U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115029344A (en) * | 2022-06-16 | 2022-09-09 | 通用生物(安徽)股份有限公司 | Preparation method of high-standard endotoxin-free plasmid |
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