CN105004806B - Concertina type spiral combination type immune affinity column - Google Patents
Concertina type spiral combination type immune affinity column Download PDFInfo
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- CN105004806B CN105004806B CN201510396422.XA CN201510396422A CN105004806B CN 105004806 B CN105004806 B CN 105004806B CN 201510396422 A CN201510396422 A CN 201510396422A CN 105004806 B CN105004806 B CN 105004806B
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- affinity column
- immune affinity
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- outlet
- end cap
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Abstract
The invention discloses a kind of concertina type spiral combination type immune affinity column.The present invention includes upper plug head, lower end cap and loading pipe.The upper plug head circumferentially carries external screw thread, and upper plug head is coordinated with loading pipe import screw thread；The lower end cap circumferentially carries female thread, and lower end cap is coordinated with loading pipe outlet screw thread；The loading pipe includes import, post chamber and outlet, and import circumferentially carries female thread, for combining the outlet of fixed upper plug head or another immune affinity column of combination fixation；Outlet is circumferential to carry external screw thread, for combining the import of fixed lower end cap or another immune affinity column of combination fixation.The present invention improves active liquid product, enhances the overall tightness of immune affinity column, in the antibody activity liquid for ensureing to have abundance while, appearance situations such as reached anti-spilled, leakage again.
Description
Technical field
The present invention relates to a kind of immune affinity column, especially a kind of concertina type spiral combination type immune affinity column.
Background technology
Used as a Sample Pretreatment Technique, solid-phase extraction column has obtained laboratory and has widely applied.With Solid-Phase Extraction
The continuous development of technology, a kind of novel extraction post --- immune affinity column are just quietly replacing conventional solid extraction column.Affine in immunity
Post（IAC）Based on antigen antibody reaction, using biomacromolecule with to a class biomacromolecule specific recognition and Reversible binding
Characteristic and a kind of separating and extracting post for making, select specificity and good adsorption cleaning performance with unique.Processed
Sample extracting solution when immune affinity column is slow transitted through, antibodies in analyte in sample extracting solution and post, and not
Remove after other combined impurity is scrubbed, then cylinder, collect analyte are rinsed with eluent.Compared with conventional solid extraction column,
Immune affinity column is higher to the selectivity of object, and absorption purifying effect is more preferable, and the reagent for being consumed is less.
Though immune affinity column has good using effect, immune affinity column there are problems that following at this stage：
1. immune affinity column due to using antibody matrix replace conventional solid filler, be holding post interior antibody activity, need by
Antibody is placed in active liquid.And the usual small volume of immune affinity column, it is impossible to there are enough storage volumetric activity liquid, and active liquid
Jing is commonly present situations such as spilling, leakage.
2. the antibody levels of single immune affinity column are fixed, and this also determines the accommodated mesh of single immune affinity column
Mark thing concentration is limited.When object exceedes the antigen limit that post interior antibody can be combined, object will be caused to be lost in, it is impossible to
Effective quantitative analyses target concentration.
3., during using immune affinity column, often appearance analysis sample liquid is more muddy to cause showing for affinity column blocking
As affecting the antibody antigen binding ability of affinity column, or even cannot normally carry out the subsequent steps such as drip washing, eluting.
Above-mentioned factor limits promoting the use of and commercialization for immune affinity column, needs a kind of simple, effective device
Solve these problems..
The content of the invention
The present invention is directed to the deficiencies in the prior art, there is provided a kind of concertina type spiral combination type immune affinity column.
The present invention solves the technical scheme taken of technical problem：
The present invention includes upper plug head, lower end cap and loading pipe.
The upper plug head circumferentially carries external screw thread, and upper plug head is coordinated with loading pipe import screw thread；The lower end cap perimeter band
There is female thread, lower end cap is coordinated with loading pipe outlet screw thread.
The loading pipe includes import, post chamber and outlet, and import circumferentially carries female thread, for combine fixed upper plug head or
The outlet of another immune affinity column of person's combination fixation；Outlet is circumferential to carry external screw thread, for combining fixed lower end cap or combination
The import of another immune affinity column of fixation.
The post chamber includes breathing zone and affine in immunity area.Breathing zone is arranged at below import, and affine in immunity area is located at out
Above mouthful, upper sieve plate, antibody fill area and lower sieve plate in affine in immunity area, are provided with from top to bottom；It is identified with most in post cavity outer wall
High liquid level (HLL) graduation mark, the highest liquid level graduation mark are located at below breathing zone.
The beneficial effects of the present invention is：
1. concertina type spiral combination type immune affinity column due to post chamber capacity it is big, and be furnished with upper and lower two plug and twist in turnover
Mouthful, active liquid product is improve, the overall tightness of immune affinity column is enhanced, is ensureing have the antibody activity liquid of abundance same
When, appearance situations such as reached anti-spilled, leakage again.
2. concertina type spiral combination type immune affinity column not only can single affinity column be independently operated, and also many immunity parent
Can be applied in combination by spiral with post, the antibody levels of immune affinity column is increased into multiple, fundamentally solve immune parent
The problem limited with the accommodated target concentration of post.
3. concertina type spiral combination type immune affinity column carries breathing zone, upper can block up during using immune affinity column
Head tightens import, breathing zone can be extruded or compressed downwards by side, increases post cavity pressure, reaches raising post chamber solution stream
Speed and the fully purpose of exclusion debris, while can also prevent post chamber from blocking.
Description of the drawings
Fig. 1 is schematic structural view of the invention；
Fig. 2 uses schematic diagram for the present invention；
Fig. 3 is applied in combination figure for the present invention.
Specific embodiment
The invention will be further described with reference to the accompanying drawings and examples.
As shown in figure 1, concertina type spiral combination type immune affinity column mainly includes following part：Upper plug head 1, lower end cap 2,
Loading pipe 3.1 high 1.5-1.8 cm of upper plug head, tool turn up screw thread, external diameter 0.8-0.9 cm, and screw type screwing is in loading pipe import；Under
2 high 1.6-2.0 cm of plug, have the screw thread that caves in, and internal diameter 0.4-0.5 cm, screw type screwing are exported in loading pipe.Loading pipe 3 is high
9.0-10.0 cm, outer wall materials are polyethylene, including import 4, post chamber 5 and outlet 6.4 high 0.9-1.0 cm of import, tool cave in
Screw thread, internal diameter are 0.9-1.0 cm, external diameter 1.0-1.1 cm, for combining fixed upper plug head 1 or another immunity of combination fixation
The outlet 6 of affinity column.6 high 0.9-1.0 cm are exported, has the screw thread that turns up, internal diameter 0.1-0.2 cm, external diameter 0.3-0.4 cm are used for
With reference to the import 4 of fixed lower end cap 2 or another immune affinity column of combination fixation.Post chamber 5 high 7.0-8.0 cm, internal diameter 1.4-
1.5 cm, external diameter 1.5-1.6 cm, can accommodate analysis volume for 6-7 mL, and post chamber 5 includes breathing zone 7, affine in immunity area 8 and most
High liquid level (HLL) graduation mark 9.It is 1.1-1.2 cm that breathing zone 7 is arranged at from import 4 away from the height that for 0.7-0.8 cm, do not stretch, thickness
It is partially thin, it is 0.1-0.2 cm.Affine in immunity area 8 is located above outlet 6, and height 1.3-1.4 cm are provided with sieve from top to bottom
Plate 10, antibody fill area 11 and lower sieve plate 12.Highest liquid level graduation mark 9 is located at post cavity outer wall and has 7 lower section 0.9-1.0 of breathing zone
cm。
Using method of the present invention is as follows：
According to the objectives thing of analysis object, corresponding antibody matrix is filled, specifically：Put in void column chamber first
Enter lower sieve plate, then insert antibody matrix, place into upper sieve plate, pushing upper sieve plate makes antibody matrix be evenly distributed, and play solid
Determine the effect of antibody fill area.
Then pouring into antibody activity liquid keeps antibody activity, the highest liquid level of active liquid that the highest of immune affinity column is less than
Liquid level graduation mark, tightens lower end cap, is placed in suitable ambient temperature to be used.According to the concentration range of object, select single
Root immune affinity column or many are applied in combination, and determine the radical of immune affinity column.The immune affinity column that will be used takes out, and goes
Fall upper lower end cap, fixed immune affinity column is in correct position（If many are applied in combination, immune affinity column is preferentially combined）.Wait to live
Property liquid entirely flow out after, needed to add the sample to be analysed liquid of suitable volumes according to experiment（If analysis sample is more muddy, can tighten
Plug, extrudes breathing zone）.After confirmation analysis sample liquid entirely flows out, appropriate leacheate is added, and extracted and discard.Finally plus
Enter suitable object eluent, collect and survey the eluent that immune affinity column flows out.
Embodiment：The affine in immunity purification applications of Fugu ocellatus toxin in aquatic products.
Fugu ocellatus toxin antibody is filled into into post intracavity, is highly 1.2-1.3 cm, covered sieve plate, and inject active liquid, twisted
Tightly upper lower end cap, is stored in 0-5 DEG C of environment, to be used.
Accurately weighing fully 2.00 g of homogenizing black scraper fish sample, in 50 mL tool plug centrifuge tubes, adds 10 mL to contain 1%
The methanol solution of acetic acid, 2 min of vortex oscillation, 60 DEG C of water bath sonicators extract 15 min, after being cooled to room temperature, 6000 r/min from
5 min of the heart.5 mL supernatant are pipetted into another 50 mL centrifuge tubes, is added 20 ml PBS solutions to be diluted, is used 1 mol/L
Sodium hydroxide solution adjusts pH to 7 ~ 8, to be clean.
Immune affinity column is taken out, removes upper lower end cap, being released after preserve in post active liquid, upper sample liquid.After end of the sample,
With 8 mL water wash affinity columns, debris in post are extracted, and discards all of the above effluent, finally contain 1% acetic acid with 4 mL
Methanol solution eluting, the eluent of collection are dried up in 60 DEG C of nitrogen, are dissolved with 1 mL mobile phases constant volume, Jing after 0.22 μm of filter membrane
For LC-MS analysis.According to above-mentioned steps parallel assay 6 times, as a result as shown in table 1.Add water in 1.50 g/kg
Under flat, the average recovery rate of six measure is 96.7%, and relative standard deviation is 3.24%.Whole affine in immunity purification process takes
For 30 min.
Residual quantity result of the table 1 using Fugu ocellatus toxin in affine in immunity purifying measuring aquatic products（n=6）
Test result indicate that using present invention energy active adsorption object, so as to reach the effect of purification.It is simultaneously whole net
Change process time short, it is simple to operate, with higher actual application value.
Above disclosed is only presently preferred embodiments of the present invention, and the right of the present invention can not be limited certainly with this
Scope, therefore the equivalent variations made according to application claims, still fall within the scope covered by the present invention.
Claims (10)
1. concertina type spiral combination type immune affinity column, including upper plug head, lower end cap and loading pipe, it is characterised in that：
The upper plug head circumferentially carries external screw thread, and upper plug head is coordinated with loading pipe import screw thread；
The lower end cap circumferentially carries female thread, and lower end cap is coordinated with loading pipe outlet screw thread；
The loading pipe includes import, post chamber and outlet, and import circumferentially carries female thread, for combining fixed upper plug head or group
Close the outlet of another immune affinity column of fixation；Outlet is circumferential to carry external screw thread, fixes for combining fixed lower end cap or combination
The import of another immune affinity column；
The post chamber includes breathing zone and affine in immunity area；Breathing zone is arranged at below import, and affine in immunity area is located in outlet
Side, is provided with upper sieve plate, antibody fill area and lower sieve plate from top to bottom in affine in immunity area；Highest liquid is identified with post cavity outer wall
Face graduation mark, the highest liquid level graduation mark are located at below breathing zone.
2. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：Described lower end cap is high
For 1.6-2.0 cm, internal diameter is 0.4-0.5 cm.
3. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：Described upper plug head is high
For 1.5-1.8 cm, external diameter is 0.8-0.9 cm.
4. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The a height of 9.0- of loading pipe
10.0 cm, outer wall materials are polyethylene.
5. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The import is a height of
0.9-1.0 cm, internal diameter are 0.9-1.0 cm, and external diameter is 1.0-1.1 cm.
6. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The outlet is a height of
0.9-1.0 cm, internal diameter are 0.1-0.2 cm, and external diameter is 0.3-0.4 cm.
7. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The post chamber can accommodate
Analysis volume is 6-7 mL, and a height of 7.0-8.0 cm in post chamber, internal diameter are 1.4-1.5 cm, and external diameter is 1.5-1.6 cm.
8. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The breathing zone is arranged
In away from import 0.7-0.8 cm, being highly 1.1-1.2 cm during compression, thickness is 0.1-0.2 cm.
9. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：Affine in immunity area height
For 1.3-1.4 cm.
10. concertina type spiral combination type immune affinity column according to claim 1, it is characterised in that：The highest liquid level
Graduation mark is located at below breathing zone at 0.9-1.0 cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510396422.XA CN105004806B (en) | 2015-07-08 | 2015-07-08 | Concertina type spiral combination type immune affinity column |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510396422.XA CN105004806B (en) | 2015-07-08 | 2015-07-08 | Concertina type spiral combination type immune affinity column |
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| Publication Number | Publication Date |
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| CN105004806A CN105004806A (en) | 2015-10-28 |
| CN105004806B true CN105004806B (en) | 2017-04-05 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201510396422.XA Expired - Fee Related CN105004806B (en) | 2015-07-08 | 2015-07-08 | Concertina type spiral combination type immune affinity column |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4126436A1 (en) * | 1990-09-17 | 1992-03-19 | Abion Ohg | DISPOSABLE REACTION TUBE FOR SOLID-PHASE IMMUNAL ANALYTICS AND METHOD FOR MEASURING COMPONENTS DETERMINABLE BY IMMUNE REACTIONS |
| CN2102789U (en) * | 1991-04-26 | 1992-04-29 | 西安医科大学 | Combined dry-pillar layer pillar |
| CN1731182A (en) * | 2005-09-02 | 2006-02-08 | 王家红 | Chloramphenicol affinity column and preparation method and use thereof |
| CN102078705A (en) * | 2010-12-28 | 2011-06-01 | 苏州先伟纳米技术有限公司 | Tandem nano fiber solid phase extraction column |
| CN102500135A (en) * | 2011-10-24 | 2012-06-20 | 利穗科技(苏州)有限公司 | Fast dynamic compression chromatography column |
| CN203154873U (en) * | 2013-04-20 | 2013-08-28 | 云南农业大学 | Combined chromatographic column |
| CN104324523A (en) * | 2014-12-01 | 2015-02-04 | 吉首大学 | Complete thread type sampling chromatographic column |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2010300A4 (en) * | 2006-02-09 | 2011-08-03 | Waters Technologies Corp | Mobil bead configuration immunoaffinity column and methods of use |
-
2015
- 2015-07-08 CN CN201510396422.XA patent/CN105004806B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4126436A1 (en) * | 1990-09-17 | 1992-03-19 | Abion Ohg | DISPOSABLE REACTION TUBE FOR SOLID-PHASE IMMUNAL ANALYTICS AND METHOD FOR MEASURING COMPONENTS DETERMINABLE BY IMMUNE REACTIONS |
| CN2102789U (en) * | 1991-04-26 | 1992-04-29 | 西安医科大学 | Combined dry-pillar layer pillar |
| CN1731182A (en) * | 2005-09-02 | 2006-02-08 | 王家红 | Chloramphenicol affinity column and preparation method and use thereof |
| CN102078705A (en) * | 2010-12-28 | 2011-06-01 | 苏州先伟纳米技术有限公司 | Tandem nano fiber solid phase extraction column |
| CN102500135A (en) * | 2011-10-24 | 2012-06-20 | 利穗科技(苏州)有限公司 | Fast dynamic compression chromatography column |
| CN203154873U (en) * | 2013-04-20 | 2013-08-28 | 云南农业大学 | Combined chromatographic column |
| CN104324523A (en) * | 2014-12-01 | 2015-02-04 | 吉首大学 | Complete thread type sampling chromatographic column |
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| CN105004806A (en) | 2015-10-28 |
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Granted publication date: 20170405 Termination date: 20180708 |