CN110426520A - A kind of magnetic particle storing liquid and its preparation - Google Patents
A kind of magnetic particle storing liquid and its preparation Download PDFInfo
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- CN110426520A CN110426520A CN201910528254.3A CN201910528254A CN110426520A CN 110426520 A CN110426520 A CN 110426520A CN 201910528254 A CN201910528254 A CN 201910528254A CN 110426520 A CN110426520 A CN 110426520A
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- magnetic particle
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
Abstract
The invention discloses a kind of magnetic particle storing liquid and its preparations, which is characterized in that the component of the magnetic particle storing liquid includes: trishydroxymethylaminomethane, sodium chloride, polyethylene glycol, Proclin-300, polysorbas20, trehalose, bovine serum albumin(BSA).The present invention by the combinations of multiple formulations, realize magnetic particle reagent sensitivity, measurement range, repeatability, in terms of, property can be promoted.By the way that bovine serum albumin(BSA), trehalose is added, the conjugate on magnetic particle is protected；Proclin-300 is added as preservative, it is suppressed that bacterial growth avoids reagent contaminated, and performance is affected；PEG 20000 is added, maintains the stability of bead suspension, is steadily carried out conducive to chemiluminescence reaction, ensure that the repeatability of test result.In summary factor, the present invention ensure that the permanently effective of magnetic particle reagent while promoting reagent performance itself.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of magnetic particle storing liquid and its preparation.
Background technique
Magnetism particulate immuno chemistry luminescence method is to rise the new analysis method of the one kind to come into vogue in recent years.Magnetic microparticle chemiluminescence
Method combines magnetic separation technique, chemiluminescence, immuno analytical method, using magnetic particle as carrier, passes through detection chemistry hair
The luminous intensity of light reaction calculates the concentration of checking matter.Because of high sensitivity, high specificity, measurement range it is wide, it is easy to operate,
The advantages that pollution-free, shows one's talent from a series of immuno analytical methods, becomes more and more popular.
However the storage of magnetic particle is to realize that a big difficulty of Magnetism particulate immuno chemistry luminescence method, the performance of magnetic particle tend to shadow
The stability for arriving entire reagent is rung, to impact to reagent performance.
In addition, different reagents object detected is different, used detection method changes therewith, and different antibody resists
There are also differences for requirement of the original to magnetic bead, magnetic bead storing liquid.
Summary of the invention
Due to above-mentioned deficiency in the prior art, the present invention devises one for magnetic bead used in Magnetism particulate immuno chemistry luminescence method
Kind storing liquid, storage, transport, use for magnetic bead in magnetic microparticle chemiluminescence reagent.
The present invention reduces background value, has correspondinglyd increase chemistry while promoting the luminescent properties of chemiluminescence reaction
Furthermore the sensitivity of luminescence reagent, the range of linearity ensure that the stability of magnetic particle storage, extend having for magnetic particle storing liquid
The effect phase.
The present invention is suitable for magnetic particle used in magnetic microparticle chemiluminescence reagent, and it is anti-to be particularly directed to not couple biotin label
The Streptavidin magnetic particle of body is better than other magnetic particle storing liquids using effect of the present invention.
The present invention provides a kind of magnetic particle storing liquid to achieve the above object, and the invention adopts the following technical scheme:
The component of the magnetic particle storing liquid includes:
Trishydroxymethylaminomethane, sodium chloride, polyethylene glycol, Proclin-300, polysorbas20, trehalose, bovine serum albumin
It is white.
In the technical program, by the combination of multiple formulations, realize magnetic particle storing liquid sensitivity, measurement range,
Repeatability, stability etc., property can be promoted.For example, by the way that bovine serum albumin(BSA), trehalose is added, protect magnetic micro-
Conjugate on grain；Be added Proclin-300 as preservative, it is suppressed that bacterial growth, avoid reagent be contaminated, performance by
It influences；PEG 20000 is added, maintains the stability of bead suspension, is steadily carried out conducive to chemiluminescence reaction, protects
The repeatability of test result is demonstrate,proved.In summary factor, the present invention ensure that magnetic is micro- while promoting reagent performance itself
Grain storing liquid it is permanently effective.
Preferably, the content of the component is as follows:
The trishydroxymethylaminomethane of 0.2wt%~0.5wt%, the sodium chloride of 0.7wt%~1.1wt%, 0.05w t%
Proclin-300,0.05vol%~0.15vol% tween of the polyethylene glycol of~2wt%, 0.02vol%~0.2vol%
20, the bovine serum albumin(BSA) of the trehalose of 0.1wt%, 0.2wt%~1wt%, remaining content complement to 100% with water.
Further, the content of the component is preferably as follows:
The trishydroxymethylaminomethane of 0.3wt%, the sodium chloride of 0.88wt%, 0.1wt% polyethylene glycol 2 0000,
The polysorbas20 of Proclin-300,0.05vol% of 0.05vol%, the trehalose of 0.1wt%, 0.5w t% bovine serum albumin
White, remaining content complements to 100% with water.
Preferably, the color of the magnetic particle storing liquid is pale yellow transparent, without insoluble matter.
Preferably, the pH range of the magnetic particle storing liquid is 7.1~7.3, and the water is ultrapure water.
The present invention also provides a kind of preparations of magnetic particle storing liquid, and its step are as follows:
S1, weigh trishydroxymethylaminomethane, sodium chloride, PEG 20000 be slowly added to containing water preparation hold
It is stirred in device to being completely dissolved；
S2, it Proclin-300, polysorbas20 is drawn is added into the resulting solution of step S1 and stir to being completely dissolved；
S3, the pH value that solution is adjusted in solution obtained in hydrochloric acid to the step S2 is drawn with pipettor；
S4, it weighs in solution obtained in bovine serum albumin(BSA) and the BSA addition step S3, blender is stirred to complete
Dissolution；
S5, it will be mixed after the resulting solution constant volume of the step S4 with water；
S6, it will be filtered to obtain magnetic particle storing liquid by the prepared solution of step S1-S5.
Preferably, the molar concentration of the hydrochloric acid is 5mol/L.
Preferably, described that the pH value that solution is adjusted in solution obtained in hydrochloric acid to the step S2 is drawn with pipettor
To 7.2 ± 0.1.
In the technical program, chemiluminescence reaction needs are carried out at a certain range of PH, and pH value had both been protected to 7.2 ± 0.1
Going on smoothly for chemiluminescence reaction has been demonstrate,proved, has also avoided the performance of magnetic particle because strong acid and strong base is destroyed.
Preferably, filter sizes needed for the filtering are 0.22 μm.
The beneficial effects of the present invention are:
1, the present invention is by the combinations of multiple formulations, realize magnetic particle reagent sensitivity, measurement range, repeatability,
Stability etc., property can be promoted.By the way that bovine serum albumin(BSA), trehalose is added, the conjugate on magnetic particle is protected；
Proclin-300 is added as preservative, it is suppressed that bacterial growth avoids reagent contaminated, and performance is affected；Poly- second is added
Glycol 20000 maintains the stability of bead suspension, steadily carries out conducive to chemiluminescence reaction, ensure that test result
Repeatability.In summary factor, the present invention ensure that the long-term of magnetic particle reagent while promoting reagent performance itself
Effectively.
2, the present invention reduces background value, has correspondinglyd increase change while promoting the luminescent properties of chemiluminescence reaction
The sensitivity of luminescence reagent is learned, furthermore the range of linearity ensure that the stability of magnetic particle storage, extend having for magnetic particle reagent
The effect phase.
3, the present invention is suitable for magnetic particle used in magnetic microparticle chemiluminescence reagent, is particularly directed to not couple biotin label
The Streptavidin magnetic particle of antibody is better than other magnetic particle storing liquids using effect of the present invention.
Specific embodiment
The invention will now be further described with reference to specific embodiments, but these examples are merely exemplary, it is not right
The scope of the present invention constitutes any restrictions.Those skilled in the art, which should be understood that, is not departing from the present invention
Under the premise of principle, several improvements and modifications can also be made, these modifications and embellishments should also be considered as the scope of protection of the present invention.
The component and content of magnetic particle storing liquid are as follows: the trishydroxymethylaminomethane of 0.2wt%~0.5wt%,
The sodium chloride of 0.7wt%~1.1wt%, the polyethylene glycol of 0.05wt%~2wt%, 0.02vol%~0.2vol%
Proclin-300,0.05vol%~0.15vol% polysorbas20, the trehalose of 0.1wt%, 0.2wt%~1wt% ox blood
Pure albumen, remaining content complement to 100% with water.
The present invention also provides a kind of preparation steps of magnetic particle storing liquid, and described steps are as follows:
S1, it trishydroxymethylaminomethane, sodium chloride, PEG 20000, trehalose is weighed is slowly added to containing water
With being stirred in container to being completely dissolved；
S2, it Proclin-300, polysorbas20 is drawn is added into the resulting solution of step S1 and stir to being completely dissolved；
S3, the pH value that solution is adjusted in solution obtained in hydrochloric acid to the step S2 is drawn with pipettor；
S4, it weighs bovine serum albumin(BSA) and is added in solution obtained in the step S3, blender is stirred to being completely dissolved；
S5, it will be mixed after the resulting solution constant volume of the step S4 with water；
S6, it will be filtered to obtain magnetic particle storing liquid by the prepared solution of step S1-S5.
Embodiment 1
For preparing 100mL:
60mL ultrapure water is taken to be put into in container using graduated cylinder；
0.3029g trishydroxymethylaminomethane, 0.8775g sodium chloride, 0.1g polyethylene glycol are weighed using precision balance
20000,0.1g trehalose is slowly added to in container, and blender is stirred to being completely dissolved；
It is added with liquid-transfering gun absorption 0.05mL Proclin-300,0.05mL polysorbas20 to in container, blender stirs
It mixes to being completely dissolved；
Suitable 5mol/L salt acid for adjusting pH is measured to 7.2 ± 0.1 with pipettor；
It weighs 0.5g bovine serum albumin(BSA) BSA using precision balance to be added to in container, blender is stirred to complete
Dissolution；
Solution is settled to 100mL with ultrapure water, is mixed；
Prepared solution is filtered using filter, filter sizes are 0.22 μm；
Solution colour is observed, should be pale yellow transparent, without insoluble matter.
The present embodiment by the combinations of multiple formulations, realize magnetic particle reagent sensitivity, measurement range, repeatability,
Stability etc., property can be promoted.By the way that BSA, trehalose is added, the conjugate on magnetic particle is protected, joined
Proclin-300 avoids bacterial growth as preservative, influences performance, joined polyethylene glycol, maintains bead suspension
Stability steadily carries out conducive to chemiluminescence reaction.In summary factor, the present invention are promoting the same of reagent performance itself
When, it ensure that the permanently effective of magnetic particle reagent.
Comparative example 1
The potassium dihydrogen phosphate of 0.02wt%, the disodium hydrogen phosphate of 0.29wt%, 0.8wt% sodium chloride,
The Proclin-300 of the potassium chloride of 0.02wt%, 0.1vol%, remaining content complement to 100% with water.
For preparing 100mL:
1) 80mL ultrapure water is taken to be put into in container using graduated cylinder；
2) using precision balance weigh 0.02g potassium dihydrogen phosphate, 0.29g disodium hydrogen phosphate, 0.80g sodium chloride,
0.02g potassium chloride is slowly added to in container, and stirring is to being completely dissolved；
3) 0.1mL Proclin-300 is measured with pipettor to be added in container, stir and evenly mix；
4) suitable 5mol/L NaOH is measured with pipettor adjust pH to 7.4 ± 0.1；
5) solution is settled to 100mL with ultrapure water, stirred and evenly mixed 15 minutes；
6) prepared solution is filtered using vacuum filter；
7) solution colour is observed, should be colorless and transparent, without insoluble matter.
Following table is embodiment 1 and the luminous value of comparative example 1 and the contrast difference of background value of various concentration, and detecting step is such as
Under: magnetic particle storing liquid and Streptavidin magnetic particle are configured to magnetic particle working solution, collocation Shanghai Yu has at biotechnology
C reactive protein (CRP) assay kit (Magnetism particulate immuno chemistry luminescence method) of limit company production, uses Nanjing enlightening Gneuss biology skill
The hair of the c reactive protein calibration object of the ACL2800 Full-automatic chemiluminescence analyzer test various concentration of art Co., Ltd production
Light value, the results are shown in Table 1.Wherein, background value is the substance for measuring 0 concentration；CV value indicates the coefficient of variation
(coefficient ofvariation), also known as coefficient of dispersion (coefficient ofdispersion) or relative deviation
(rsd), it is the ratio between standard deviation and average value, is indicated with percentage, calculation formula are as follows: CV=sd/mean × 100%.mean
It is expressed as the average value of luminous value.Sd is standard deviation.
Table 1
As shown in Table 1, using chemical illuminating reagent of the invention, other magnetic particle storing liquids are compared, background value is low, shines
Performance is good, and high level calibration object and the luminous value ratio of low value calibration object are bigger.
The present invention reduces background value, has correspondinglyd increase chemistry while promoting the luminescent properties of chemiluminescence reaction
The sensitivity of luminescence reagent, the range of linearity.
Embodiment 2
This specific embodiment discloses another proportion, and preparation step is same as Example 1, the ratio of ingredient are as follows:
The trishydroxymethylaminomethane of 0.2wt%, the sodium chloride of 0.7wt%, the polyethylene glycol of 0.05wt%, 0.02vol%
The polysorbas20 of Proclin-300,0.05vol%, the trehalose of 0.1wt%, 0.2wt% bovine serum albumin(BSA), it is remaining to contain
Amount complements to 100% with water.
Embodiment 3
This specific embodiment discloses another proportion, and preparation step is same as Example 1, the ratio of ingredient are as follows:
The trishydroxymethylaminomethane of 0.5wt%, the sodium chloride of 1.1wt%, the polyethylene glycol of 2wt%, 0.2vol% Proclin-
300, the bovine serum albumin(BSA) of the polysorbas20 of 0.15vol%, the trehalose of 0.1wt%, 1wt%, remaining content are supplied with water
To 100%.
Following table is embodiment 2 and the luminous value of comparative example 3 and the contrast difference of background value of various concentration, and detecting step is such as
Under: magnetic particle storing liquid and Streptavidin magnetic particle are configured to magnetic particle working solution, collocation Shanghai Yu has at biotechnology
C reactive protein (CRP) assay kit (Magnetism particulate immuno chemistry luminescence method) of limit company production, uses Nanjing enlightening Gneuss biology skill
The c reactive protein calibration object of the ACL2800 Full-automatic chemiluminescence analyzer test various concentration of art Co., Ltd production,
The results are shown in Table 2.
Table 2
As shown in Table 2, embodiment 2 is lower than comparative example 1 with the background value of embodiment 3, and luminous value is higher than comparative example 1, so
Background value can be reduced, the spirit of chemical illuminating reagent has been correspondinglyd increase while promoting the luminescent properties of chemiluminescence reaction
Sensitivity, the range of linearity.
Embodiment 4
This specific embodiment, which discloses, configures magnetic particle storing liquid prepared by embodiment 1 and Streptavidin magnetic particle
At magnetic particle working solution, 4 DEG C, 37 DEG C of storages are respectively put into, were taken out in the 0th, 2,5,8 day, arranged in pairs or groups raw with a collection of c reactive protein
Object element labelled antibody, acridinium ester label antibody are tested with a collection of c reactive protein calibration object, compare luminous value difference, result is such as
Shown in table 3:
Table 3
As shown in Table 3, magnetic particle storing liquid prepared by embodiment 1, it is micro- that collocation Streptavidin magnetic particle is configured to magnetic
Grain working solution, be with the 0th day luminous value difference of storage difference when calibration object concentration is 1mg/L after being stored 8 days at 4 DEG C-
3%；When calibration object concentration is 5mg/L, difference is -5%；When calibration object concentration is 80mg/L, difference is 5%.
Embodiment 5
This specific embodiment discloses magnetic particle storing liquid prepared by comparative example 1 and is configured to Streptavidin magnetic particle
Magnetic particle working solution is respectively put into 4 DEG C, 37 DEG C of storages, took out in the 0th, 2,5,8 day, arranges in pairs or groups with a collection of c reactive protein biology
Plain labelled antibody, acridinium ester label antibody are tested with a collection of c reactive protein calibration object, compare luminous value difference, result is such as
Shown in table 4:
Table 4
As shown in Table 4, it is luminous with storage the 0th day after magnetic particle storing liquid prepared by comparative example 1 is stored 8 days at 4 DEG C
The different difference when calibration object concentration is 1mg/L of value difference is -13%；When calibration object concentration is 5mg/L, difference is -14%；In school
Difference is -15% when quasi- product concentration is 80mg/L.
By the comparison of table 3 and table 4 it is found that magnetic particle storing liquid is compared with prepared by comparative example 1 prepared by embodiment 1
Magnetic particle storing liquid is little in 8 days difference of 37 DEG C of 8 days luminous values of storage and 4 DEG C of storages, and 4 DEG C are stored 8 days and storage the 0th day
Luminous value difference is little, and stability is good.
Embodiment 6
This specific embodiment discloses embodiment 1 and comparative example 1, and the method for inspection is as follows: by magnetic particle storing liquid and chain
Mould Avidin magnetic particle is configured to magnetic particle working solution, and arrange in pairs or groups Procalcitonin biotin labelled antibodies, acridinium ester label antibody make
Various concentration is tested with the 800 Full-automatic chemiluminescence analyzer of ACL2 that Nanjing enlightening Gneuss Bioisystech Co., Ltd produces
Procalcitonin calibration object, result is as shown in table 5.
Table 5
As shown in Table 5, using chemical illuminating reagent of the invention, other magnetic particle storing liquids are compared, background value is low, shines
Performance is good, and high level calibration object and the luminous value ratio of low value calibration object are bigger.
In summary, the present invention is suitable for magnetic particle used in most of Magnetism particulate immuno chemistry luminescence method, is particularly directed to not even
The Streptavidin magnetic particle for joining biotin labelled antibodies is better than other magnetic particle storing liquids using effect of the present invention.
Embodiments of the present invention above described embodiment only expresses, the description thereof is more specific and detailed, but can not
Therefore limitations on the scope of the patent of the present invention are interpreted as.It should be pointed out that for those of ordinary skill in the art,
Without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection model of the invention
It encloses.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (9)
1. a kind of magnetic particle storing liquid, which is characterized in that the component of the magnetic particle storing liquid includes:
Trishydroxymethylaminomethane, sodium chloride, polyethylene glycol, Proclin-300, polysorbas20, trehalose, bovine serum albumin(BSA).
2. a kind of magnetic particle storing liquid as described in claim 1, which is characterized in that the content of the component is as follows:
The trishydroxymethylaminomethane of 0.2wt%~0.5wt%, the sodium chloride of 0.7wt%~1.1wt%, 0.05wt%~
Proclin-300,0.05vol%~0.15vol% tween of the polyethylene glycol of 2wt%, 0.02vol%~0.2vol%
20, the bovine serum albumin(BSA) of the trehalose of 0.1wt%, 0.2wt%~1wt%, remaining content complement to 100% with water.
3. a kind of magnetic particle storing liquid as described in claim 1, which is characterized in that including following component and content:
The trishydroxymethylaminomethane of 0.3wt%, the sodium chloride of 0.88wt%, 0.1wt% PEG 20000,
The polysorbas20 of Proclin-300,0.05vol% of 0.05vol%, the trehalose of 0.1wt%, 0.5wt% bovine serum albumin
White, remaining content complements to 100% with water.
4. a kind of magnetic particle storing liquid as described in claim 1, it is characterised in that:
The color of the magnetic particle storing liquid is pale yellow transparent, without insoluble matter.
5. a kind of magnetic particle storing liquid as described in claim 1, it is characterised in that:
The pH range of the magnetic particle storing liquid is 7.1~7.3, and the water is ultrapure water.
6. a kind of preparation of magnetic particle storing liquid includes a kind of magnetic particle storing liquid as described in Claims 1 to 5, feature exists
In, including following preparation step:
S1, weigh trishydroxymethylaminomethane, sodium chloride, PEG 20000 be slowly added to containing water in container
Stirring is to being completely dissolved；
S2, it Proclin-300, polysorbas20 is drawn is added into the resulting solution of step S1 and stir to being completely dissolved；
S3, the pH value that solution is adjusted in solution obtained in hydrochloric acid to the step S2 is drawn with pipettor；
S4, it weighs bovine serum albumin(BSA) and is added in solution obtained in the step S3, blender is stirred to being completely dissolved；
S5, it will be mixed after the resulting solution constant volume of the step S4 with water；
S6, it will be filtered to obtain magnetic particle storing liquid by the prepared solution of step S1-S5.
7. a kind of preparation of magnetic particle storing liquid as claimed in claim 6, which is characterized in that in the step S3:
The molar concentration of the hydrochloric acid is 5mol/L.
8. a kind of preparation of magnetic particle storing liquid as claimed in claim 6, which is characterized in that in the step S3:
Described drawn with pipettor adjusts the pH value of solution to 7.2 ± 0.1 in solution obtained in hydrochloric acid to the step S2.
9. a kind of preparation of magnetic particle storing liquid as claimed in claim 6, which is characterized in that in the step S6:
Filter sizes needed for the filtering are 0.22 μm.
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| CN109725148A (en) * | 2018-12-30 | 2019-05-07 | 广东环凯微生物科技有限公司 | A kind of immunomagnetic beads preservation liquid |
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| CN112710824A (en) * | 2020-12-15 | 2021-04-27 | 北京美联泰科生物技术有限公司 | Buffer solution for preserving superparamagnetic particles and protein connectors thereof and preparation method thereof |
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Application publication date: 20191108 |