CN105004806A - Telescopic spiral combination type immunoaffinity column - Google Patents
Telescopic spiral combination type immunoaffinity column Download PDFInfo
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- CN105004806A CN105004806A CN201510396422.XA CN201510396422A CN105004806A CN 105004806 A CN105004806 A CN 105004806A CN 201510396422 A CN201510396422 A CN 201510396422A CN 105004806 A CN105004806 A CN 105004806A
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- immune affinity
- affinity column
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- combined type
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Abstract
The invention discloses a telescopic spiral combination type immunoaffinity column. The telescopic spiral combination type immunoaffinity column comprises an upper plug, a lower plug and a sample loading tube. The circumference of the upper plug is provided with an outside thread, and the upper plug cooperates with the inlet of the sample loading tube in a threaded manner; the circumference of the lower plug is provided with inside thread, and the lower plug cooperates with the outlet of the sample loading tube in a threaded manner; the sample loading tube comprises the inlet, a column cavity and the outlet, and the circumference of the inlet is provided with an inside threaded used for combining fixation of the upper plug or combining fixation of the outlet of an another immunoaffinity column; and the circumference of the outlet is provided with an outside thread used for combining fixation of the lower plug or combining fixation of the inlet of the another immunoaffinity column. The telescopic spiral combination type immunoaffinity column improves the volume of an active solution, has enhanced integral sealing property, guarantees sufficient antibody active solution and prevents overflow and solution leakage.
Description
Technical field
The present invention relates to a kind of immune affinity column, especially a kind of concertina type spiral combined type immune affinity column.
Background technology
As a Sample Pretreatment Technique, solid-phase extraction column has obtained laboratory and has applied widely.Along with the development of solid phase extraction techniques, a kind of novel extraction post---immune affinity column just quietly replaces conventional solid extraction column.Immune affinity column (IAC), based on antigen-antibody reaction, utilizes biomacromolecule to have a kind of separating and extracting post made the characteristic of a class biomacromolecule specific recognition and Reversible binding, has unique selection selectivity and good adsorption cleaning performance.The sample extracting solution processed is when slow transitting through immune affinity column, and the antibody of analysis thing in post in sample extracting solution is combined, and other not combined impurity removing after washing, then rinse cylinder with eluent, collect analyte.Comparatively conventional solid extraction column, immune affinity column is stronger to the selectivity of object, and absorption purifying effect is better, and the reagent consumed is less.
Though immune affinity column has good result of use, present stage immune affinity column there is following problem:
1. immune affinity column replaces conventional solid filler owing to using antibody matrix, for keeping antibody activity in post, antibody need be placed in active liquid.And the usual small volume of immune affinity column, enough storage volumetric active liquids cannot be had, and often there is the situations such as spilling, leakage in active liquid.
2. the antibody levels of single immune affinity column is fixing, and this held target concentration also determining single immune affinity column is limited.When object exceed antibody in post can in conjunction with antigen limit, object will be caused to run off, cannot effective quantitative test target concentration.
3., in the process using immune affinity column, normal appearance analyzes the more muddy phenomenon causing affinity column to block of sample liquid, affects the antibody antigen binding ability of affinity column, even cannot normally carry out the subsequent step such as drip washing, wash-out.
Above-mentioned factor limits promoting the use of of immune affinity column and commercialization, needs a kind of simple, effective device to solve these problems.。
Summary of the invention
The present invention is directed to the deficiencies in the prior art, provide a kind of concertina type spiral combined type immune affinity column.
The technical scheme that technical solution problem of the present invention is taked is:
The present invention includes upper plug head, lower end cap and loading pipe.
Described upper plug head is circumferential with external thread, upper plug head and loading pipe import threaded engagement; Described lower end cap circumference is with internal thread, and lower end cap and loading pipe export threaded engagement.
Described loading pipe comprises import, post chamber and outlet, and import is circumferential with internal thread, for fixing the outlet of another immune affinity column in conjunction with fixing upper plug head or combination; Outlet is circumferential with external thread, for fixing the import of another immune affinity column in conjunction with fixing lower end cap or combination.
Described post chamber comprises breathing zone and immune affinity regions.Breathing zone is arranged at below import, and immune affinity regions is positioned at outlet top, is provided with upper sieve plate, antibody fill area and lower sieve plate in immune affinity regions from top to bottom; Have most high liquid level (HLL) scale mark in post cavity outer wall mark, this most high liquid level (HLL) scale mark is positioned at below breathing zone.
Beneficial effect of the present invention is:
1. concertina type spiral combined type immune affinity column due to post chamber capacity large, and be furnished with upper and lower two plugs and twist in import and export, improve active liquid volume, enhance the overall tightness of immune affinity column, ensure sufficient antibody activity liquid simultaneously, reach again the appearance of the situations such as anti-spilled, leakage.
2. concertina type spiral combined type immune affinity column not only can independently use by single affinity column, and many immune affinity columns combinationally use by spiral, make the antibody levels of immune affinity column become multiple to increase, fundamentally solve the problem that the held target concentration of immune affinity column is limited.
3. concertina type spiral combined type immune affinity column is with breathing zone, in use immune affinity column process, upper plug head can be tightened import, extrude by side or compress breathing zone downwards, increase post cavity pressure, reach the object improving post chamber solution flow rate and fully get rid of Liquid Residue, also can prevent post chamber from blocking simultaneously.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Fig. 2 is that the present invention uses schematic diagram;
Fig. 3 is that the present invention combinationally uses figure.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
As shown in Figure 1, concertina type spiral combined type immune affinity column mainly comprises with lower part: upper plug head 1, lower end cap 2, loading pipe 3.The high 1.5-1.8 cm of upper plug head 1, tool turns up screw thread, external diameter 0.8-0.9 cm, and screw type screwing is in the import of loading pipe; The high 1.6-2.0 cm of lower end cap 2, tool caves in screw thread, and internal diameter 0.4-0.5 cm, screw type screwing exports in loading pipe.The high 9.0-10.0 cm of loading pipe 3, outer wall materials is tygon, comprises import 4, post chamber 5 and outlet 6.The high 0.9-1.0 cm of import 4, tool caves in screw thread, and internal diameter is 0.9-1.0 cm, external diameter 1.0-1.1 cm, for fixing the outlet 6 of another immune affinity column in conjunction with fixing upper plug head 1 or combination.Export 6 high 0.9-1.0 cm, tool turns up screw thread, internal diameter 0.1-0.2 cm, external diameter 0.3-0.4 cm, for fixing the import 4 of another immune affinity column in conjunction with fixing lower end cap 2 or combination.The high 7.0-8.0 cm in post chamber 5, internal diameter 1.4-1.5 cm, external diameter 1.5-1.6 cm, can hold analysis volume is 6-7 mL, and post chamber 5 comprises breathing zone 7, immune affinity regions 8 and most high liquid level (HLL) scale mark 9.Breathing zone 7 is arranged at from import 4 apart from being 0.7-0.8 cm, and not flexible is highly 1.1-1.2 cm, and thickness is partially thin, is 0.1-0.2 cm.Immunity affinity regions 8 is positioned at above outlet 6, and height 1.3-1.4 cm, is provided with upper sieve plate 10, antibody fill area 11 and lower sieve plate 12 from top to bottom.Most high liquid level (HLL) scale mark 9 is positioned at 0.9-1.0 cm below post cavity outer wall tool breathing zone 7.
Using method of the present invention is as follows:
According to the objectives thing of analytic target, fill corresponding antibody matrix, specifically: first in void column chamber, put into lower sieve plate, then antibody matrix is inserted, put into upper sieve plate again, press down upper sieve plate and antibody matrix is evenly distributed, and play the effect of sessile antibody fill area.
Then pour into antibody activity liquid and keep antibody activity, the most high liquid level (HLL) of active liquid lower than the most high liquid level (HLL) scale mark of immune affinity column, will be tightened lower end cap, is placed in suitable environment temperature to be used.According to the concentration range of object, select single immune affinity column or many to combinationally use, determine the radical of immune affinity column.The immune affinity column that will use takes out, and removes upper lower end cap, and fixing immune affinity column is in correct position (if many combinationally use, preferentially combining immune affinity column).After active liquid flows out entirely, experimentally need the sample liquid to be analyzed (if it is more muddy to analyze sample, upper plug head can be tightened, extruding breathing zone) adding suitable volumes.After confirmation analysis sample liquid flows out entirely, add appropriate leacheate, and extracted and discard.Finally add suitable object eluent, the eluent that collection analysis immune affinity column flows out.
Embodiment: the affine purification applications of the immunity of tetraodotoxin in aquatic products.
Tetraodotoxin antibody is filled in post chamber, is highly 1.2-1.3 cm, builds sieve plate, and inject active liquid, tighten lower end cap, be stored in 0-5 DEG C of environment, to be used.
Accurately take fully that homogeneous black scraper fish sample 2.00 g is in 50 mL tool plug centrifuge tubes, add the methanol solution that 10 mL contain 1% acetic acid, vortex oscillation 2 min, 60 DEG C of water bath sonicator extract 15 min, after being cooled to room temperature, and centrifugal 5 min of 6000 r/min.Pipette 5 mL supernatants in another 50 mL centrifuge tube, add 20 ml PBS solution and dilute, regulate pH to 7 ~ 8 with 1 mol/L sodium hydroxide solution, to be clean.
Take out immune affinity column, remove upper lower end cap, release after preserving active liquid in post, upper sample liquid.After end of the sample, with 8 mL water wash affinity columns, extract Liquid Residue in post, and discard above whole efflux, the methanol solution wash-out of 1% acetic acid is finally contained with 4 mL, the eluent collected dries up in 60 DEG C of nitrogen, dissolves with 1 mL mobile phase constant volume, for LC-MS analysis after 0.22 μm of filter membrane.According to above-mentioned steps replicate determination 6 times, result is as shown in table 1.Under 1.50 μ g/kg Pitch-based sphere, the average recovery rate measured for six times is 96.7%, and relative standard deviation is 3.24%.Whole immunity is affine purification process is consuming time is 30 min.
Table 1 utilize tetraodotoxin in immune affine purifying measuring aquatic products residual quantity result (
n=6)
Experimental result shows to use the present invention can effective adsorbed target thing, thus reaches the effect of purification.The whole purification process time is short, simple to operate simultaneously, has higher actual application value.
Above disclosedly be only preferred embodiment of the present invention, certainly can not limit the interest field of the present invention with this, therefore according to the equivalent variations that application claims is done, still belong to the scope that the present invention is contained.
Claims (10)
1. concertina type spiral combined type immune affinity column, comprises upper plug head, lower end cap and loading pipe, it is characterized in that:
Described upper plug head is circumferential with external thread, upper plug head and loading pipe import threaded engagement;
Described lower end cap circumference is with internal thread, and lower end cap and loading pipe export threaded engagement;
Described loading pipe comprises import, post chamber and outlet, and import is circumferential with internal thread, for fixing the outlet of another immune affinity column in conjunction with fixing upper plug head or combination; Outlet is circumferential with external thread, for fixing the import of another immune affinity column in conjunction with fixing lower end cap or combination;
Described post chamber comprises breathing zone and immune affinity regions; Breathing zone is arranged at below import, and immune affinity regions is positioned at outlet top, is provided with upper sieve plate, antibody fill area and lower sieve plate in immune affinity regions from top to bottom; Have most high liquid level (HLL) scale mark in post cavity outer wall mark, this most high liquid level (HLL) scale mark is positioned at below breathing zone.
2. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: described lower end cap height is 1.6-2.0 cm, and internal diameter is 0.4-0.5 cm.
3. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: described upper plug head height is 1.5-1.8 cm, and external diameter is 0.8-0.9 cm.
4. concertina type spiral combined type immune affinity column according to claim 1, it is characterized in that: loading Guan Gaowei 9.0-10.0 cm, outer wall materials is tygon.
5. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: described import height is 0.9-1.0 cm, and internal diameter is 0.9-1.0 cm, and external diameter is 1.0-1.1 cm.
6. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: described outlet height is 0.9-1.0 cm, and internal diameter is 0.1-0.2 cm, and external diameter is 0.3-0.4 cm.
7. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: it is 6-7 mL that described post chamber can hold analysis volume, and post chamber height is 7.0-8.0 cm, and internal diameter is 1.4-1.5 cm, and external diameter is 1.5-1.6 cm.
8. concertina type spiral combined type immune affinity column according to claim 1, it is characterized in that: described breathing zone is arranged at from import 0.7-0.8 cm place, be highly 1.1-1.2 cm during compression, thickness is 0.1-0.2 cm.
9. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: immune affinity regions height is 1.3-1.4 cm.
10. concertina type spiral combined type immune affinity column according to claim 1, is characterized in that: described most high liquid level (HLL) scale mark is positioned at 0.9-1.0 cm place below breathing zone.
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 |
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| CN201510396422.XA CN105004806B (en) | 2015-07-08 | 2015-07-08 | Concertina type spiral combination type immune affinity column |
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| CN105004806A true CN105004806A (en) | 2015-10-28 |
| CN105004806B CN105004806B (en) | 2017-04-05 |
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Citations (8)
| 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 |
| US20090269859A1 (en) * | 2006-02-09 | 2009-10-29 | Waters Investments Limited | Mobile Bead Configuration Immunoaffinity Column and Methods of Use |
| 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 |
-
2015
- 2015-07-08 CN CN201510396422.XA patent/CN105004806B/en not_active Expired - Fee Related
Patent Citations (8)
| 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 |
| US20090269859A1 (en) * | 2006-02-09 | 2009-10-29 | Waters Investments Limited | Mobile Bead Configuration Immunoaffinity Column and Methods of Use |
| 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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| CN105004806B (en) | 2017-04-05 |
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Granted publication date: 20170405 Termination date: 20180708 |