CN108593907A - A kind of dilution and preparation method thereof of carboxylated magnetic particle - Google Patents
A kind of dilution and preparation method thereof of carboxylated magnetic particle Download PDFInfo
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Abstract
The invention discloses a kind of dilutions and preparation method thereof of carboxylated magnetic particle, are matched and are formed in parts by weight by following components：39~55 parts of N (2 ethoxy) piperazine N'2 ethane sulfonic acids, 28~38 parts of dipotassium hydrogen phosphate, 9~18 parts of disodium ethylene diamine tetraacetate, 11~29 parts of zinc chloride, 7~12 parts of magnesium chloride, 6~13 parts of heparin sodium, 15~22 parts of sucrose, 0.1~0.3 part of hydrochloric acid, 4~12 parts of glucose, 1~3 part of bovine serum albumin(BSA), 0.2~0.6 part of mice serum, 0.3~0.7 part of glycerine, 0.2~0.4 part of Proclin300,0.4~0.8 part of polysorbas20,100~300 parts of deionized water.The present invention utilizes the synergistic effect between each component, prepare the dilution of carboxylated magnetic particle, the dilution is for diluting carboxylated magnetic particle mother liquor into working solution so that the working solution can long-term preservation, and the performances such as the specificity of reagent, stability, repeatability are unaffected.
Description
Technical field
The present invention relates to the dilution of chemo-immunity luminescence reagent field more particularly to a kind of carboxylated magnetic particle and its
Preparation method.
Background technology
With the development of science and technology, while chemiluminescence advance is advanced.It is developed so far, chemiluminometry
Rapid development has been obtained in biomedical sector, mainly because its analytic process does not need additional light source, can avoid detecting
Other light sources interfere in journey, to improve its sensitivity.Fixing means is particularly important in chemiluminescence immune assay, mainly will
Enzyme, antigen, antibody etc. are wrapped on certain high molecular materials, suitable solid phase carrier be improve immunoassay method stability,
The one of the most effective ways of the performances such as sensitivity, specificity.Wherein carboxylated magnetic particle is suitable for antibody and combination occurs instead
It answers, while large specific surface area, combined amount is more；However center of gravity is mainly placed in the research and development of magnetic particle by many companies, and neglect
After working solution being made into depending on magnetic particle, important function of buffer system during reagin-antigen-reactive, to influence
The performance indicator of reagent.Therefore, it is particularly important to prepare a kind of dilution work of carboxylated magnetic particle.
Invention content
Present invention solves the technical problem that：In order to obtain a kind of dilution of carboxylated magnetic particle, the diluted
After working solution is made in carboxylated magnetic particle, magnetic particle is not influenced and is combined with antibody/antigen, and working solution is suitble to long-term protect
It deposits, ensures that reagent specification is unaffected.
Technical solution：A kind of dilution of carboxylated magnetic particle, is matched by following components and is formed in parts by weight：N-
39~55 parts of (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, 28~38 parts of dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate 9~18
Part, 11~29 parts of zinc chloride, 7~12 parts of magnesium chloride, 6~13 parts of heparin sodium, 15~22 parts of sucrose, 0.1~0.3 part of hydrochloric acid, Portugal
4~12 parts of grape sugar, 1~3 part of bovine serum albumin(BSA), 0.2~0.6 part of mice serum, 0.3~0.7 part of glycerine, Proclin
300 0.2~0.4 part, 0.4~0.8 part of Tween-20,100~300 parts of deionized water.
A kind of preferred dilution of carboxylated magnetic particle, is matched by following components and is formed in parts by weight：N- (2- hydroxyls
Ethyl) 51 parts of piperazine-N'-2- ethane sulfonic acids, 32 parts of dipotassium hydrogen phosphate, 12 parts of disodium ethylene diamine tetraacetate, 23 parts of zinc chloride, chlorine
Change 9 parts of magnesium, 11 parts of heparin sodium, 18 parts of sucrose, 0.2 part of hydrochloric acid, 6 parts of glucose, 2 parts of bovine serum albumin(BSA), mice serum 0.4
Part, 0.5 part of glycerine, 0.3 part of Proclin300,0.5 part of Tween-20,200 parts of deionized water.
A kind of dilution liquid and preparation method thereof of carboxylated magnetic particle, includes the following steps：
1st step, by N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate according to
In secondary addition deionized water, and with 120~160rpm mixing speeds, stirring to solution is clarified to get solution A；
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added stirring with 80~150rpm by the 2nd step
In the deionized water for mixing speed stirring, 30~60min is to get solution B for stirring；
3rd step mixes solution A with solution B, and 30~60min is stirred with the mixing speed of 50~90rpm, adjust pH to
6.6~7.5, and be filtered to get solution C with 0.22 μm of filter membrane；
Bovine serum albumin(BSA), mice serum are added in deionized water 4th step, and 20 are stirred with the speed of 80~120rpm
~30min, and be filtered to get solution D with 0.45 μm of filter membrane；
5th step, by solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with 50~
The mixing speed of 100rpm stirs 70~120min, you can a kind of dilution of carboxylated magnetic particle is made.
Preferably, by N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, ethylenediamine tetrem in the 1st step
Acid disodium sequentially adds in deionized water, and with 150rpm mixing speeds, and stirring to solution is clarified to get solution A.
Preferably, zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added with 120rpm in the 2nd step
Mixing speed stirring deionized water in, stirring 40min to get solution B.
Preferably, solution A is mixed in the 3rd step with solution B, 40min is stirred with the mixing speed of 70rpm, adjust pH to
7.2, and be filtered to get solution C with 0.22 μm of filter membrane.
Preferably, bovine serum albumin(BSA), mice serum are added in deionized water in the 4th step, are stirred with the speed of 100rpm
25min is mixed, and is filtered to get solution D with 0.45 μm of filter membrane.
Preferably, by solution C and solution D mixing in the 5th step, and glycerine, Proclin 300, Tween-20 is added, with
The mixing speed of 70rpm stirs 100min, you can a kind of dilution of carboxylated magnetic particle is made.
Advantageous effect：The present invention utilizes the synergistic effect between each component, prepares the dilution of carboxylated magnetic particle
Liquid, the dilution is for diluting carboxylated magnetic particle mother liquor into working solution so that the working solution can long-term preservation, and reagent
The performances such as specificity, stability, repeatability are unaffected.
Specific implementation mode
Embodiment 1
A kind of dilution of carboxylated magnetic particle, is matched by following components and is formed in parts by weight：N- (2- ethoxys)
51 parts of piperazine-N'-2- ethane sulfonic acids, 32 parts of dipotassium hydrogen phosphate, 12 parts of disodium ethylene diamine tetraacetate, 23 parts of zinc chloride, magnesium chloride 9
Part, 11 parts of heparin sodium, 18 parts of sucrose, 0.2 part of hydrochloric acid, 6 parts of glucose, 2 parts of bovine serum albumin(BSA), 0.4 part of mice serum, the third three
0.5 part of alcohol, 0.3 part of Proclin300,0.5 part of Tween-20,200 parts of deionized water.
A kind of dilution liquid and preparation method thereof of carboxylated magnetic particle, includes the following steps：
1st step, by N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate according to
In secondary addition deionized water, and with 150rpm mixing speeds, stirring to solution is clarified to get solution A；
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added the stirring speed with 120rpm by the 2nd step
It spends in the deionized water of stirring, 40min is to get solution B for stirring；
3rd step mixes solution A with solution B, and 40min is stirred with the mixing speed of 70rpm, adjusts pH to 7.2, and with
0.22 μm of filter membrane is filtered to get solution C；
Bovine serum albumin(BSA), mice serum are added in deionized water 4th step, and 25min is stirred with the speed of 100rpm,
And it is filtered to get solution D with 0.45 μm of filter membrane；
5th step, by solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with stirring for 70rpm
Speed is mixed, 100min is stirred, you can a kind of dilution of carboxylated magnetic particle is made.
Embodiment 2
A kind of dilution of carboxylated magnetic particle, is matched by following components and is formed in parts by weight：N- (2- ethoxys)
45 parts of piperazine-N'-2- ethane sulfonic acids, 31 parts of dipotassium hydrogen phosphate, 12 parts of disodium ethylene diamine tetraacetate, 21 parts of zinc chloride, magnesium chloride 9
Part, 8 parts of heparin sodium, 19 parts of sucrose, 0.2 part of hydrochloric acid, 8 parts of glucose, 1 part of bovine serum albumin(BSA), 0.5 part of mice serum, the third three
0.4 part of alcohol, 300 0.2 parts of Proclin, 0.5 part of Tween-20,160 parts of deionized water.
A kind of dilution liquid and preparation method thereof of carboxylated magnetic particle, includes the following steps：
1st step, by N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate according to
In secondary addition deionized water, and with 120~160rpm mixing speeds, stirring to solution is clarified to get solution A；
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added the stirring speed with 110rpm by the 2nd step
It spends in the deionized water of stirring, 50min is to get solution B for stirring；
3rd step mixes solution A with solution B, stirs 55min with the mixing speed of 50~90rpm, adjusts pH to 6.8,
And it is filtered to get solution C with 0.22 μm of filter membrane；
Bovine serum albumin(BSA), mice serum are added in deionized water 4th step, and 26min is stirred with the speed of 90rpm, and
It is filtered to get solution D with 0.45 μm of filter membrane；
5th step, by solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with stirring for 80rpm
Speed is mixed, 100min is stirred, you can a kind of dilution of carboxylated magnetic particle is made.
Embodiment 3
A kind of dilution of carboxylated magnetic particle, is matched by following components and is formed in parts by weight：N- (2- ethoxys)
50 parts of piperazine-N'-2- ethane sulfonic acids, 31 parts of dipotassium hydrogen phosphate, 15 parts of disodium ethylene diamine tetraacetate, 21 parts of zinc chloride, magnesium chloride
10 parts, 9 parts of heparin sodium, 19 parts of sucrose, 0.2 part of hydrochloric acid, 8 parts of glucose, 3 parts of bovine serum albumin(BSA), 0.5 part of mice serum, third
0.6 part of triol, 300 0.4 parts of Proclin, 0.7 part of Tween-20,300 parts of deionized water.
A kind of dilution liquid and preparation method thereof of carboxylated magnetic particle, includes the following steps：
1st step, by N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate according to
In secondary addition deionized water, and with 150rpm mixing speeds, stirring to solution is clarified to get solution A；
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added the stirring speed with 130rpm by the 2nd step
It spends in the deionized water of stirring, 40min is to get solution B for stirring；
3rd step mixes solution A with solution B, stirs 45min with the mixing speed of 50~90rpm, adjusts pH to 7.2,
And it is filtered to get solution C with 0.22 μm of filter membrane；
Bovine serum albumin(BSA), mice serum are added in deionized water 4th step, and 27min is stirred with the speed of 100rpm,
And it is filtered to get solution D with 0.45 μm of filter membrane；
5th step, by solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with stirring for 80rpm
Speed is mixed, 90min is stirred, you can a kind of dilution of carboxylated magnetic particle is made.
Embodiment 4
The dilution for the carboxylated magnetic particle that Examples 1 to 3 is prepared is dilute for carboxylated magnetic particle mother liquor
It is interpreted into working solution, with corresponding antibody, antigen combination at reagent set, investigates the thermal stability of the reagent, repeatability, blank
Can, it is as a result as follows：
| Thermal stability (37 DEG C accelerate 7 days) | Repeated (%) | Blank (counts) | |
| Embodiment 1 | 95.1% | 5.6 | 5372 |
| Embodiment 2 | 90.5% | 7.6 | 7531 |
| Embodiment 3 | 93.2% | 6.9 | 7999 |
Claims (8)
1. a kind of dilution of carboxylated magnetic particle, which is characterized in that matched and formed in parts by weight by following components：N-
39~55 parts of (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, 28~38 parts of dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate 9~18
Part, 11~29 parts of zinc chloride, 7~12 parts of magnesium chloride, 6~13 parts of heparin sodium, 15~22 parts of sucrose, 0.1~0.3 part of hydrochloric acid, Portugal
4~12 parts of grape sugar, 1~3 part of bovine serum albumin(BSA), 0.2~0.6 part of mice serum, 0.3~0.7 part of glycerine, Proclin
3000.2~0.4 parts, 0.4~0.8 part of Tween-20,100~300 parts of deionized water.
2. a kind of dilution of carboxylated magnetic particle according to claim 1, which is characterized in that by following components by weight
Measure number proportioning composition：51 parts of N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, 32 parts of dipotassium hydrogen phosphate, ethylenediamine tetra-acetic acid
12 parts of disodium, 23 parts of zinc chloride, 9 parts of magnesium chloride, 11 parts of heparin sodium, 18 parts of sucrose, 0.2 part of hydrochloric acid, 6 parts of glucose, cow's serum
2 parts of albumin, 0.4 part of mice serum, 0.5 part of glycerine, 0.3 part of Proclin300,0.5 part of Tween-20, deionized water 200
Part.
3. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle described in claim 1, which is characterized in that including following step
Suddenly：
1st step adds N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate successively
Enter in deionized water, and with 120~160rpm mixing speeds, stirring to solution is clarified to get solution A；
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added the stirring speed with 80~150rpm by the 2nd step
It spends in the deionized water of stirring, 30~60min is to get solution B for stirring；
3rd step mixes solution A with solution B, and 30~60min is stirred with the mixing speed of 50~90rpm, adjust pH to 6.6~
7.5, and be filtered to get solution C with 0.22 μm of filter membrane；
Bovine serum albumin(BSA), mice serum are added in deionized water 4th step, with the speed of 80~120rpm stirring 20~
30min, and be filtered to get solution D with 0.45 μm of filter membrane；
5th step, by solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with 50~100rpm's
Mixing speed stirs 70~120min, you can a kind of dilution of carboxylated magnetic particle is made.
4. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle according to claim 3, which is characterized in that in the 1st step
N- (2- ethoxys) piperazine-N'-2- ethane sulfonic acids, dipotassium hydrogen phosphate, disodium ethylene diamine tetraacetate are sequentially added into deionized water
In, and with 150rpm mixing speeds, stirring to solution is clarified to get solution A.
5. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle according to claim 3, which is characterized in that in the 2nd step
Zinc chloride, magnesium chloride, heparin sodium, sucrose, hydrochloric acid, glucose are sequentially added with the deionization of the mixing speed stirring of 120rpm
In water, 40min is to get solution B for stirring.
6. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle according to claim 3, which is characterized in that in the 3rd step
Solution A is mixed with solution B, 40min is stirred with the mixing speed of 70rpm, adjusts pH to 7.2, and carry out with 0.22 μm of filter membrane
Filtering is to get solution C.
7. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle according to claim 3, which is characterized in that in the 4th step
Bovine serum albumin(BSA), mice serum are added in deionized water, 25min is stirred with the speed of 100rpm, and with 0.45 μm of filter membrane
It is filtered to get solution D.
8. a kind of dilution liquid and preparation method thereof of carboxylated magnetic particle according to claim 3, which is characterized in that in the 5th step
By solution C and solution D mixing, and glycerine, Proclin 300, Tween-20 is added, with the mixing speed of 70rpm, stirring
100min, you can a kind of dilution of carboxylated magnetic particle is made.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254145A (en) * | 2018-11-08 | 2019-01-22 | 郑州安图生物工程股份有限公司 | For improving the dilution of matrix effect between fresh serum and third party's Quality Control |
| CN111781346A (en) * | 2020-07-17 | 2020-10-16 | 成都赛普克生物科技股份有限公司 | Enzyme-linked immune diluent for whole blood and preparation method and using method thereof |
| CN111812313A (en) * | 2020-06-22 | 2020-10-23 | 北京生物制品研究所有限责任公司 | Dissociation method of antigen in aluminum adjuvant adsorption type novel coronavirus inactivated vaccine |
| CN111812326A (en) * | 2020-07-23 | 2020-10-23 | 四川携光生物技术有限公司 | Kit for quantitatively detecting content of PD-L1 and PD-1 conjugate |
| CN112014567B (en) * | 2020-08-20 | 2023-11-17 | 金华科生物技术河北有限公司 | Kit for measuring hypersensitive C reaction protein |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111781346B (en) * | 2020-07-17 | 2023-02-03 | 成都赛普克生物科技股份有限公司 | Enzyme-linked immune diluent for whole blood and preparation method and using method thereof |
| CN111812326A (en) * | 2020-07-23 | 2020-10-23 | 四川携光生物技术有限公司 | Kit for quantitatively detecting content of PD-L1 and PD-1 conjugate |
| CN112014567B (en) * | 2020-08-20 | 2023-11-17 | 金华科生物技术河北有限公司 | Kit for measuring hypersensitive C reaction protein |
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Application publication date: 20180928 |