CN103048448A - Stationary liquid - Google Patents
Stationary liquid Download PDFInfo
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- CN103048448A CN103048448A CN2012105611290A CN201210561129A CN103048448A CN 103048448 A CN103048448 A CN 103048448A CN 2012105611290 A CN2012105611290 A CN 2012105611290A CN 201210561129 A CN201210561129 A CN 201210561129A CN 103048448 A CN103048448 A CN 103048448A
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- stationary liquid
- trehalose
- elisa
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Abstract
A stationary liquid belongs to the technical field of in vitro diagnostic reagents. The stationary liquid takes a PBS buffer solution as a solvent, and cane sugar and trehalose as solute, wherein as for the PBS buffer solution, the molarity of phosphate group is 0.01 mol/L, and the pH value is 7.2-7.4; and in the stationary liquid, the mass percent of the cane sugar is 5-30 percent, and the mass percent of the trehalose is 5-30 percent. Compared with the prior art, the stationary liquid has the advantages that the stationary liquid is used for fixing the activity of the protein capturing an antibody ELISA, so as to finally prepare an ELISA-precoated plate which can be preserved for a long time at the temperature of 4 DEG C; and the saving time and stability of an ELISA kit are increased, so as to reduce the scrapped rate of the ELIS kit due to the reduction and loss of the activity of the pre-coated plate; and accordingly, the economic loss is reduced.
Description
Technical field
The present invention is a kind of immobile liquid, belongs in vitro diagnostic reagent technical field.
Background technology
Enzyme-linked Immunosorbent Assay (ELISA) experiment is highly sensitive because of it, and sample obtains easily, has simple to operately become internationally recognized protein quantification goldstandard.One of gordian technique of ELISA kit preparation is exactly the preparation of antibody solid phase carrier ELISA Plate.The preparation method of pre-coated elisa plate routine discards coating buffer coating buffer is hatched certain hour in enzyme mark hole after, places 4 ℃ can preserve 1 month, places-20 ℃, can preserve 3 months; Add drying agent again, vacuumize processing, can preserve about 1 year at 4 ℃, but this can not satisfy the needs of ELISA kit long preservation, the ELISA pre-coated elisa plate needs the longer protein active retention time.
Be 200910192897.1 such as the patent No., the preparation method that name is called a kind of streptavidin pre-coated elisa plate with use in disclosed method, the preparation process of traditional E LISA pre-coated elisa plate is: preparation coating buffer, coated, dry.In recent years, after coated step, the sealing step is arranged, dry again after the sealing; But at present, between sealing and drying steps, do not have to use the immobile liquid that is specifically designed to the ankyrin activity, so that the holding time falls short of, cause the ELISA kit to scrap because pre-coated elisa plate is expired, produce no small economic loss.
Summary of the invention
The object of the present invention is to provide a kind of ELISA pre-coated elisa plate that can make at the immobile liquid of 4 degree preservations more than 3 years.
The present invention solve the problems of the technologies described above the technical scheme that adopts be this immobile liquid with the PBS damping fluid as solvent, sucrose and trehalose are as solute; The phosphate radical volumetric molar concentration of described PBS damping fluid is 0.01mol/L, and pH=7.2-7.4, the massfraction of described sucrose are 5%-30%, and the massfraction of described trehalose is 5%-30%.
As preferably, described PBS pH of buffer=7.3, the massfraction of sucrose is 30%, the massfraction of trehalose is 30%.
The advantage that the present invention compared with prior art has is: be fixed with the protein active of immobile liquid to capture antibody ELISA, the ELISA pre-coated elisa plate that finally makes can be under 4 degree conditions long preservation.Increase holding time and the stability of ELISA kit, reduced the ELISA kit and scrapped because of pre-coated elisa plate activity decreased or forfeiture, reduced economic loss.
Embodiment
The present invention is described in further detail below in conjunction with embodiment, and following examples are explanation of the invention and the present invention is not limited to following examples.
The preparation process of embodiment 1:Human IL-6 pre-coated elisa plate:
1, preparation coating buffer: will resist Human IL-6 capture antibody is 0.01mol/L with the phosphate radical volumetric molar concentration, and the PBS damping fluid of pH=7.2 is diluted to 2ug/ml, obtains coating buffer.
2, coated: as in enzyme mark hole, to add the coating buffer of 100ul, 4 ℃ of coated spending the night;
3, sealing: abandon coating buffer, add 1%BSA-PBST confining liquid 300ul/ hole, room temperature sealing 4 hours;
4, preparation immobile liquid: be 0.01mol/L in the phosphate radical volumetric molar concentration, add sucrose and trehalose in the PBS damping fluid of pH=7.3, the massfraction that makes sucrose is 30%, and the massfraction of trehalose is 30%, is fixed liquid.
5, fixing: abandon confining liquid, add the every hole 300ul of above-mentioned immobile liquid, room temperature is fixed 0.5 hour;
6, dry: abandon immobile liquid, ambient temperature overnight is dried.
7, add drying agent, aluminium foil bag vacuumizes, and 4 ℃ of preservations obtain Human IL-6 pre-coated elisa plate.
The phosphate radical volumetric molar concentration of the confining liquid of 1%BSA-PBST described in the present embodiment is 0.01mol/L, and the Tween20 volume fraction is that 1 ‰, BSA massfraction is 1%, pH=7.3.
Above-mentioned Human IL-6 pre-coated elisa plate can be preserved 28 days through 37 degree accelerated tests.According to the Arrhenius formula conclusion, 37 degree are preserved and were equivalent to 4 degree preservations 48 days in 1 day, therefore draw present embodiment Human IL-6 pre-coated elisa plate and can preserve more than 3 years at 4 degree.
Other embodiment: catch albumen take Human IL-6 and after example is made Human IL-6 pre-coated elisa plate, carry out the holding time test, the massfraction of sucrose and trehalose is the proportioning in the following form in the described immobile liquid, step is identical with embodiment 1, preservation fate such as following table that 37 degree accelerated tests corresponding to different proportionings record:
| Sucrose | Trehalose | Preserve fate (37 degree) |
| 5% | 5% | 23 days |
| 10% | 10% | 24 days |
| 15% | 15% | 25 days |
| 20% | 20% | 26 days |
| 25% | 25% | 27 days |
| 30% | 30% | 28 days |
According to the Arrhenius formula conclusion, 37 degree are preserved and to be equivalent to 4 degree in 1 day and to preserve 48 days, 37 minimum maintenances 23 days when spending in the table, therefore draw the Human IL-6 pre-coated elisa plate that makes with immobile liquid of the present invention and spend 4 and can preserve more than 3 years.
In addition, need to prove, the specific embodiment described in this instructions, its proportioning, title that technique is named etc. can be different.Allly conceive equivalence or the simple change that described structure, feature and principle are done according to patent of the present invention, be included in the protection domain of patent of the present invention.Those skilled in the art can make various modifications or replenish or adopt similar mode to substitute described specific embodiment; only otherwise depart from structure of the present invention or surmount this scope as defined in the claims, all should belong to protection scope of the present invention.
Although the present invention with embodiment openly as above, is not to limit protection scope of the present invention, any technician who is familiar with this technology not breaking away from change and the retouching of doing in the spirit and scope of the present invention, all should belong to protection scope of the present invention.
Claims (3)
1. immobile liquid, it is characterized in that: as solvent, sucrose and trehalose are as solute with the PBS damping fluid; The phosphate radical volumetric molar concentration of described PBS damping fluid is 0.01mol/L, and pH=7.2-7.4, the massfraction of described sucrose are 5%-30%, and the massfraction of described trehalose is 5%-30%.
2. immobile liquid according to claim 1 is characterized in that: described PBS pH of buffer=7.3.
3. immobile liquid according to claim 1 and 2, it is characterized in that: the massfraction of described sucrose is 30%, the massfraction of described trehalose is 30%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210561129.0A CN103048448B (en) | 2012-12-21 | 2012-12-21 | Stationary liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210561129.0A CN103048448B (en) | 2012-12-21 | 2012-12-21 | Stationary liquid |
Publications (2)
| Publication Number | Publication Date |
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| CN103048448A true CN103048448A (en) | 2013-04-17 |
| CN103048448B CN103048448B (en) | 2015-01-14 |
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| CN201210561129.0A Active CN103048448B (en) | 2012-12-21 | 2012-12-21 | Stationary liquid |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108872596A (en) * | 2018-07-05 | 2018-11-23 | 重庆巴而思生物科技有限公司 | A kind of ELISA detection kit of HPV16 L1 antibody |
| CN111528219A (en) * | 2020-05-13 | 2020-08-14 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN113267621A (en) * | 2021-05-14 | 2021-08-17 | 北京金诺百泰生物技术有限公司 | Stabilizer for ELISA kit coated plate, preparation method of stabilizer, kit coated plate and kit |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101095950A (en) * | 2006-06-30 | 2008-01-02 | 辽宁成大生物股份有限公司 | Hydrophobia vaccine freezing drying preparations for stable human beings and the preparations thereof |
| CN102331491A (en) * | 2011-06-20 | 2012-01-25 | 洪伟 | Plate enclosing stabilizing agent |
| CN102331494A (en) * | 2011-06-16 | 2012-01-25 | 广州艺佳生物科技有限公司 | Sealing and stabilizing agent for microporous board |
| CN102375056A (en) * | 2010-08-27 | 2012-03-14 | 烟台赛尔斯生物技术有限公司 | Immobilized bio-macromolecular stabilizer as well as preparation method and application thereof |
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2012
- 2012-12-21 CN CN201210561129.0A patent/CN103048448B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101095950A (en) * | 2006-06-30 | 2008-01-02 | 辽宁成大生物股份有限公司 | Hydrophobia vaccine freezing drying preparations for stable human beings and the preparations thereof |
| CN102375056A (en) * | 2010-08-27 | 2012-03-14 | 烟台赛尔斯生物技术有限公司 | Immobilized bio-macromolecular stabilizer as well as preparation method and application thereof |
| CN102331494A (en) * | 2011-06-16 | 2012-01-25 | 广州艺佳生物科技有限公司 | Sealing and stabilizing agent for microporous board |
| CN102331491A (en) * | 2011-06-20 | 2012-01-25 | 洪伟 | Plate enclosing stabilizing agent |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108872596A (en) * | 2018-07-05 | 2018-11-23 | 重庆巴而思生物科技有限公司 | A kind of ELISA detection kit of HPV16 L1 antibody |
| CN111528219A (en) * | 2020-05-13 | 2020-08-14 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN111528219B (en) * | 2020-05-13 | 2022-03-15 | 上海市计量测试技术研究院 | Freeze-drying protective agent for T lymphocyte subpopulation counting standard substance and application thereof |
| CN113267621A (en) * | 2021-05-14 | 2021-08-17 | 北京金诺百泰生物技术有限公司 | Stabilizer for ELISA kit coated plate, preparation method of stabilizer, kit coated plate and kit |
| CN113267621B (en) * | 2021-05-14 | 2021-12-17 | 北京金诺百泰生物技术有限公司 | Stabilizer for ELISA kit coated plate, preparation method of stabilizer, kit coated plate and kit |
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| CN103048448B (en) | 2015-01-14 |
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Effective date of registration: 20161229 Address after: 310011 Gongshu District, Hangzhou, Hunan Road, No. 36, on the 3rd floor, No. Patentee after: Hangzhou branch biotechnology Limited by Share Ltd Address before: 310011, block A, block B, No. 36, Mau Mau Road, North Hangzhou Software Park, Zhejiang, China Patentee before: Hangzhou Maotiansai Technology Co., Ltd. |