CN105300966A - Preserving fluid and preparation method thereof - Google Patents
Preserving fluid and preparation method thereof Download PDFInfo
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- CN105300966A CN105300966A CN201510791109.6A CN201510791109A CN105300966A CN 105300966 A CN105300966 A CN 105300966A CN 201510791109 A CN201510791109 A CN 201510791109A CN 105300966 A CN105300966 A CN 105300966A
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Abstract
The invention provides a preserving fluid and its preparation method. The preserving fluid comprises 0.1-10 g/L of buffer salt, 100-300 g/L of an antibody stabilizer, 0.1-1 g/L of an anticorrosive bacteriostatic agent and 2-12 g/L of carragheenan and water. In comparison with the prior art, the invention has the following advantages: the buffer salt, antibody stabilizer and anticorrosive bacteriostatic agent in the preserving fluid maintain stability of magnetic micro-particles or magnetic nano-particles and antibody in the whole system; and viscosity of the carragheenan can decrease exponentially with rising of temperature. Solubility of the carragheenan increases with rising of temperature, molecular dissociation is intensified, electrostatic attraction between half-esterfied sulfate radicals is weakened, and molecular entanglement decreases. Then, viscosity is lowered. But viscosity increases when temperature is reduced. At 30 DEG C, molecules are gradually entangled to form a net structure so as to sharply increase viscosity. Thus, the preserving fluid is in a viscous or gelatinous state at the temperature of 2-20 DEG C but is in a fluid state at the operating temperature of 35-38 DEG C.
Description
Technical field
The invention belongs to chemiluminescence immune assay field, particularly relate to a kind of conserving liquid and preparation method thereof.
Background technology
Chemiluminescence immune assay (Chemiluminescenceimmunoassay, CLIA) be that chemiluminescence system is combined with immune response, the mass signatures antibody relevant with chemiluminescence or antigen, after antigen to be measured or antibody response, through the chemiluminescent labels of separated free state, other correlatives adding chemiluminescence system produce chemiluminescence, carry out the quantitative or qualitative detection of antigen or antibody.Be developed so far from the seventies in last century, chemiluminescence immune assay has become a kind of maturation, advanced ultramicron active substance detection technique.Because it has the advantages such as highly sensitive, wide, the simple to operate automaticity of high specificity, sensing range is high, reagent stability good, pollution is very little, develop swift and violent over nearly 10 years, range of application is increasingly extensive, is particularly considered to current immune quantitative in the large application of clinical detection, environmental analysis and food security three and analyzes optimal method.
Solid phase material good in chemiluminescence immune assay can promote the activity of solidifying protein molecular, reduces non-specific adsorption, accelerates the diffusion reaction speed of albumen, for immunoreactive carrying out smoothly provides powerful guarantee.Modern immunoassay technology has quite a few to employ synthesis solid phase material as reaction carriers, and the solid phase carrier at present for immunoassay mainly contains three classes: microwell plate, particulate and porosint.Wherein, micro-size particles and nanometer particle can be dispersed in greatly in liquid phase due to the little and specific surface area of its particle diameter, diffusion reaction between antibody antigen is accelerated, magnetic particle (magnetic particle) and magnetic nanoparticle can also play fast enriching centrifugation, not only as reaction solid phase but also as the carrier be separated, its application makes immune response under the condition of intimate homogeneous phase, achieve reaction kinetics fast, simultaneously can at quick separating floating preteins and Antibody-antigen complex under magnetic fields, thus simplify operation, shorten the reaction time.
But along with chemiluminescence immunoassay technology make rapid progress while, a lot of deficiency also exposes thereupon, one of them is exactly the preservation problem of magnetic particle and magnetic nanoparticle, if the conserving liquid viscosity of magnetic particle or magnetic nano particle is too low in examination bar, first easily occur that magnetic particle or magnetic nanoparticle are reunited and sink to the bottom phenomenon, be unfavorable for the stable storage of label on particle, secondly splash possibly in transportation or in use procedure, be stained with chamber wall or top, thus the particle played a role in testing process is reduced, affect test result; Otherwise if viscosity is too high, the problem of preservation may solve, but due to viscosity too large, be unfavorable for that mixing material is dispersed and cause affect antibody mediated immunity and react.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of conserving liquid and preparation method thereof, and the viscosity of this conserving liquid can vary with temperature.
The invention provides a kind of conserving liquid, it is characterized in that, comprising:
Preferably, the thickening agent of 1 ~ 100g/L except carragheen is also comprised.
Preferably, described thickening agent except carragheen is selected from one or more in gelatin, sodium alginate, hydroxyethyl cellulose, gellan gum, sucrose, polyvinyl alcohol (PVA), polyglycol and polyvinylpyrrolidone.
Preferably, described buffer salt is one or more in sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, 3-(N-morpholinyl) propane sulfonic acid, 2-(N-morpholine) ethyl sulfonic acid monohydrate, NaOH, hydrochloric acid, sodium chloride and potassium chloride.
Preferably, described antibody stabilization agent is one or more in casein, bovine serum albumin(BSA), cow's serum and glycerine.
Preferably, described antiseptic and inhibiting bacteria function agent is one or more in sodium azide, Proclin300 and chloromycetin.
Preferably, described carragheen is one or more in κ type carragheen, ι type carragheen and λ type carragheen.
Present invention also offers a kind of preparation method of conserving liquid, comprising:
0.1 ~ 10g/L buffer salt, the agent of 100 ~ 300g/L antibody stabilization, the agent of 0.1 ~ 1g/L antiseptic and inhibiting bacteria function, 2 ~ 12g/L carragheen are mixed with water, heating for dissolving, obtains conserving liquid.
Preferably, the temperature of described heating is 35 DEG C ~ 85 DEG C.
Present invention also offers the application of conserving liquid in magnetic particle or magnetic nanoparticle are preserved.
The invention provides a kind of conserving liquid and preparation method thereof, this conserving liquid comprises 0.1 ~ 10g/L buffer salt, the agent of 100 ~ 300g/L antibody stabilization, the agent of 0.1 ~ 1g/L antiseptic and inhibiting bacteria function, 2 ~ 12g/L carragheen and water.Compared with prior art, buffer salt in conserving liquid of the present invention, antibody stabilization agent and antiseptic and inhibiting bacteria function agent are keep magnetic particle in whole system or magnetic nanoparticle and antibody stable, carragheen is by D-galactopyranose and 3, the calcium of the high component polysaccharide sulfuric ester of 6-Anhydrogalactose composition, magnesium, sodium, ammonium carnallite, its viscosity can raise and exponentially rule decline with temperature, the solubleness raising carragheen with temperature increases, molecular dissociation aggravates, weaken the electrostatic attraction between half-esterification sulfate radical, intermolecular entanglement reduces, cause viscosity, on the contrary, when temperature reduces, viscosity increases, viscosity is made sharply to increase because carrageenan molecule progressively starts to be wound reticulate texture when 30 DEG C, therefore this conserving liquid is made to present thickness or gel at 2 DEG C ~ 20 DEG C, and working temperature 35 DEG C ~ 38 DEG C in fluid state.
Accompanying drawing explanation
Fig. 1 is Promega luminous value linear equation curve map in the embodiment of the present invention 1.
Embodiment
Below in conjunction with the accompanying drawing of the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
The invention provides a kind of conserving liquid, comprising:
Wherein, the kind of described buffer salt is buffer salt well known to those skilled in the art, there is no special restriction, in the present invention, be preferably one or more in sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, 3-(N-morpholinyl) propane sulfonic acid, 2-(N-morpholine) ethyl sulfonic acid monohydrate, NaOH, hydrochloric acid, sodium chloride and potassium chloride; The content of described buffer salt is preferably 1 ~ 10g/L, is more preferably 3 ~ 8g/L; When described buffer salt comprises sodium dihydrogen phosphate, the content of described sodium dihydrogen phosphate is preferably 0.5 ~ 2.5g/L; When described buffer salt comprises sodium hydrogen phosphate, the content of described sodium hydrogen phosphate is preferably 0.2 ~ 2g/L, is more preferably 0.5 ~ 2g/L, then is preferably 1 ~ 2g/L; When described buffer salt comprises potassium dihydrogen phosphate, the content of described potassium dihydrogen phosphate is preferably 0.1 ~ 0.5g/L, is more preferably 0.1 ~ 0.3g/L; When described buffer salt comprises 3-(N-morpholinyl) propane sulfonic acid (MOPS), the content of described MOPS is preferably 0.5 ~ 3g/L, is more preferably 1 ~ 3g/L; When described buffer salt comprises 2-(N-morpholine) ethyl sulfonic acid monohydrate (MES), the content of described MES is preferably 0.5 ~ 3g/L, is more preferably 1 ~ 2.5g/L; When described buffer salt comprises NaOH, the content of described NaOH is preferably 0.1 ~ 0.5g/L, is more preferably 0.1 ~ 0.3g/L; When described buffer salt comprises hydrochloric acid, the content of described hydrochloric acid is preferably 0.1 ~ 0.5g/L; When described buffer salt comprises sodium chloride, the content of described sodium chloride is preferably 3 ~ 10g/L, is more preferably 3 ~ 6g/L; When described buffer salt comprises potassium chloride, the content of described potassium chloride is preferably 0.5 ~ 2g/L, is more preferably 0.9 ~ 2g/L.
The kind of described antibody stabilization agent is antibody stabilization agent well known to those skilled in the art, there is no special restriction, is preferably one or more in casein, bovine serum albumin(BSA) (BSA), cow's serum and glycerine in the present invention; The content of described antibody stabilization agent is preferably 100 ~ 300g/L, is more preferably 100 ~ 250g/L, then is preferably 160 ~ 220g/L; When described antibody stabilization agent comprises casein, described caseic content is preferably 30 ~ 60g/L; When described antibody stabilization agent comprises BSA, the content of described BSA is preferably 30 ~ 80g/L; When described antibody stabilization agent comprises cow's serum, the content of described cow's serum is preferably 80 ~ 150g/L; When described antibody stabilization agent comprises glycerine, the content of described glycerine is preferably 120 ~ 180g/L.
The kind of described antiseptic and inhibiting bacteria function agent is antiseptic and inhibiting bacteria function agent well known to those skilled in the art, there is no special restriction, is preferably one or more in sodium azide, Proclin300 and chloromycetin in the present invention; The content of described antiseptic and inhibiting bacteria function agent is preferably 0.3 ~ 1g/L, is more preferably 0.5 ~ 0.8g/L; When described antiseptic and inhibiting bacteria function agent comprises sodium azide, the content of described sodium azide is preferably 0.1 ~ 0.5g/L, is more preferably 0.2 ~ 0.4g/L; When described antiseptic and inhibiting bacteria function agent comprises Proclin300, the content of described Proclin300 is preferably 0.3 ~ 1g/L, is more preferably 0.3 ~ 0.7g/L; When described antiseptic and inhibiting bacteria function agent comprises chloromycetin, the content of described chloromycetin is preferably 0.1 ~ 0.5g/L, is more preferably 0.1 ~ 0.3g/L.
Described carragheen be preferably in κ type carragheen, ι type carragheen and λ type carragheen one or more, be more preferably one wherein; The content of described carragheen is preferably 3 ~ 10g/L, is more preferably 2 ~ 8g/L.
According to the present invention, described conserving liquid preferably also comprises the thickening agent of 1 ~ 100g/L except carragheen.Described thickening agent except carragheen is thickening agent well known to those skilled in the art, there is no special restriction, in the present invention, be preferably one or more in gelatin, sodium alginate, hydroxyethyl cellulose, gellan gum, sucrose, polyvinyl alcohol (PVA), polyglycol and polyvinylpyrrolidone; Described gelatin is preferably isinglass and/or Bos taurus domesticus Gmelin; Described polyglycol preferred degree of polymerization is the polyglycol of 6000 ~ 20000; Described polyvinylpyrrolidone is preferably PVP K30 and/or polyvinylpyrrolidone 360.In the present invention, when comprising gelatin in conserving liquid, the content of described gelatin is preferably 4 ~ 15g/L, is more preferably 6 ~ 15g/L; When comprising sodium alginate in conserving liquid, the content of described sodium alginate is preferably 1 ~ 10g/L, is more preferably 1 ~ 5g/L; When comprising hydroxyethyl cellulose in conserving liquid, the content of described hydroxyethyl cellulose is preferably 50 ~ 100g/L; When comprising gellan gum in conserving liquid, the content of described gellan gum is preferably 1 ~ 5g/L, is more preferably 1 ~ 3g/L; When comprising sucrose in conserving liquid, the content of described sucrose is preferably 300 ~ 400g/L; When comprising polyvinyl alcohol (PVA) in conserving liquid, the content of described polyvinyl alcohol (PVA) is preferably 1 ~ 5g/L, is more preferably 1 ~ 3g/L; When comprising polyglycol in described conserving liquid, the content of described polyglycol is preferably 1 ~ 5g/L, is more preferably 2 ~ 4g/L; When comprising polyvinylpyrrolidone in described conserving liquid, the content of described polyvinylpyrrolidone is preferably 3 ~ 10g/L, is more preferably 5 ~ 8g/L.Thickening agent except carragheen can be used as supplementary thickener or the complex thickener of carragheen, with carragheen acting in conjunction, makes the viscosity of conserving liquid comparatively large under the storage temperature of magnetic nano-particle, and less under its working temperature.
Described water is used for the volume of supplementary conserving liquid.
In the present invention, the pH value of described conserving liquid is preferably 5.5 ~ 7.5.
Buffer salt in conserving liquid of the present invention, antibody stabilization agent and antiseptic and inhibiting bacteria function agent are keep magnetic particle in whole system or magnetic nanoparticle and antibody stable, carragheen is by D-galactopyranose and 3, the calcium of the high component polysaccharide sulfuric ester of 6-Anhydrogalactose composition, magnesium, sodium, ammonium carnallite, its viscosity can raise and exponentially rule decline with temperature, the solubleness raising carragheen with temperature increases, molecular dissociation aggravates, weaken the electrostatic attraction between half-esterification sulfate radical, intermolecular entanglement reduces, cause viscosity, on the contrary, when temperature reduces, viscosity increases, viscosity is made sharply to increase because carrageenan molecule progressively starts to be wound reticulate texture when 30 DEG C, therefore this conserving liquid is made to present thickness or gel at 2 DEG C ~ 20 DEG C, and working temperature 35 DEG C ~ 38 DEG C in fluid state.
Present invention also offers a kind of preparation method of above-mentioned conserving liquid, comprising: 0.1 ~ 10g/L buffer salt, the agent of 100 ~ 300g/L antibody stabilization, the agent of 0.1 ~ 1g/L antiseptic and inhibiting bacteria function, 2 ~ 12g/L carragheen are mixed with water, heating for dissolving, obtains conserving liquid.
According to the present invention, preferably also add the thickening agent of 1 ~ 100g/L except carragheen.Wherein, described buffer salt, antibody stabilization agent, antiseptic and inhibiting bacteria function agent, carragheen, water and the thickening agent except carragheen are all same as above, do not repeat them here.
In the present invention, preferred elder generation just 0.1 ~ 10g/L buffer salt, the agent of 100 ~ 300g/L antibody stabilization, the agent of 0.1 ~ 1g/L antiseptic and inhibiting bacteria function, 2 ~ 12g/L carragheen mixes with part water, and the mode of described mixing is preferably ultrasonic or stirs, heating for dissolving, add remaining water constant volume again, obtain conserving liquid.
The temperature of described heating is preferably 35 DEG C ~ 85 DEG C, is more preferably 40 DEG C ~ 80 DEG C, then is preferably 50 DEG C ~ 80 DEG C.
Present invention also offers the application of a kind of above-mentioned conserving liquid in magnetic particle or magnetic nano-particle are preserved.
In order to further illustrate the present invention, below in conjunction with embodiment, a kind of conserving liquid provided by the invention and preparation method thereof is described in detail.
Reagent used in following examples is commercially available.
Embodiment 1
700ml aqua sterilisa is added in 1000ml conical flask, then adds 1.8gNa
2hPO
412H
2o, 0.22gKH
2pO
4, 3gNaCl, 0.3gKCl, 50gBSA, 150g glycerine, 0.3g sodium azide and 0.5gProclin300, ultrasonic mixing, filter, 40 DEG C of heating water baths, add 3g polyglycol 12000,6g carragheen κ type, 1g sodium alginate and 1g polyvinyl alcohol (PVA) to dissolving completely in batches, then add water and be settled to 1000ml, obtain conserving liquid.
By dispersed to conserving liquid for the magnetic nanoparticle of c reactive protein bag quilt.
Performance test: accurately pipetting 100 μ l, 0.1mol/LpH values with liquid-transfering gun is that the phosphate buffer (PB6.5) of 6.5 is to 1.5mL centrifuge tube, add the conserving liquid that 10 μ l0.1 μ g/mlCRP antigens or 1 μ g/mlCRP antigen, the antibody of 20 μ l alkali phosphatase enzyme marks and 100 μ l are dispersed with magnetic nanoparticle successively wherein, then with liquid-transfering gun, mixed liquor is broken up evenly, start timing.After 5min, above-mentioned centrifuge tube is placed in magnetic frame, after 30s, sucks clear liquid, wash twice with 300 μ l0.01mol/LPB7.4.With 100 μ l, 0.2mol/L, pH=9.0Tris-HCl damping fluids, washed magnetic nanoparticle is uniformly dispersed, add 50 μ lCDPStar, mix rear beginning timing, be added to 96 hole white ELISA Plate, with the luminous value of Promega semi-automatic luminometer test 3min.Repeat above-mentioned steps, prepare 3 parallel samples altogether, count No. 1, No. 2 and No. 3, obtain the results are shown in Table 1, Fig. 1 is that the Promega luminous value test result obtained carries out reason conjunction, obtain Linear equations, the unit of its horizontal ordinate is μ g/mL, and the unit of ordinate is RLU (relative light unit).
Table 1 embodiment 1Promega luminous value test result
| Promega test value (RLU) | No. 1 | No. 2 | No. 3 | Mean value |
| Blank | 1344 | 1020 | 1149 | 1171 |
| CRP antigen 0.1 μ g/mL | 9847 | 10438 | 10721 | 10335 |
| CRP antigen 1 μ g/mL | 91302 | 90527 | 94306 | 92045 |
| CRP sample 0.16 μ g/mL | 15549 | 16327 | 16284 | 16053 |
| CRP sample 1.31 μ g/mL | 120918 | 114706 | 110495 | 115373 |
Conserving liquid embodiment 1 being dispersed with to magnetic nanoparticle carries out retention test, obtains result to be: 2 DEG C ~ 8 DEG C, magnetic nanoparticle not sedimentation completely; Under 37 DEG C of environment, there is sedimentation in 3 days in magnetic nanoparticle, within 6 days, do not have complete sedimentation.
Conserving liquid embodiment 1 being dispersed with to magnetic nanoparticle carries out accelerated stability test test: get the conserving liquid that 100 μ L are dispersed with magnetic nanoparticle and add to 1.5mL centrifuge tube, make 40,20 are placed in 2 DEG C ~ 8 DEG C Refrigerator stores, and other 20 are placed in 37 DEG C of baking oven accelerated deteriorations; All the other method of testings are identical with performance test, obtain stability test and the results are shown in Table 2.
Table 2 embodiment 1 stability test result
Embodiment 2
700ml aqua sterilisa is added in 1000ml conical flask, then 2.3gMOPS, 0.19gNaOH, 2.2gNaCl, 1.3gKCl, 60g calf serum, 140g glycerine, 0.3g sodium azide and 0.2g chloromycetin is added, ultrasonic mixing, filter, 40 DEG C of heating water baths, add 5g polyvinylpyrrolidone 360,3g carragheen λ type, 3g carragheen ι type, 1g gellan gum and 1g polyvinyl alcohol (PVA) to dissolving completely, then adding water is settled to 1000ml, obtains conserving liquid in batches.
By β
2the magnetic nanoparticle of-MG bag quilt is dispersed to conserving liquid.
Performance test: accurately pipette 100 μ l, 0.1mol/LPB6.5 to 1.5mL centrifuge tube with liquid-transfering gun, adds 10 μ l0.05 μ g/ml β successively wherein
2-MG antigen or 0.5 μ g/ml β
2the antibody of-MG antigen, 20 μ l alkali phosphatase enzyme marks and 100 μ l are dispersed with the conserving liquid of magnetic nanoparticle, then break up evenly with liquid-transfering gun by mixed liquor, start timing.After 5min, above-mentioned centrifuge tube is placed in magnetic frame, after 30s, sucks clear liquid, wash twice with 300 μ l0.01mol/LPB7.4.With 100 μ l, 0.2mol/L, pH=9.0Tris-HCl damping fluids, washed magnetic nanoparticle is uniformly dispersed, add 50 μ lCDPStar, mix rear beginning timing, be added to 96 hole white ELISA Plate, with the luminous value of Promega semi-automatic luminometer test 3min.Repeat above-mentioned steps, prepare 3 parallel samples altogether, count No. 1, No. 2 and No. 3, obtain the results are shown in Table 3.
Table 3 embodiment 2Promega luminous value test result
| Promega test value (RLU) | No. 1 | No. 2 | No. 3 | Mean value |
| Blank | 987 | 1154 | 1039 | 1060 |
| β 2-MG antigen 0.05 μ g/mL | 13654 | 15279 | 13786 | 14239 |
| β 2-MG antigen 0.5 μ g/mL | 139071 | 142879 | 149546 | 143832 |
| β 2-MG sample 0.08 μ g/mL | 21792 | 22940 | 24841 | 23191 |
| β 2-MG sample 0.47 μ g/mL | 134570 | 141055 | 129776 | 135133 |
Conserving liquid embodiment 2 being dispersed with to magnetic nanoparticle carries out retention test, obtains result to be: 2 DEG C ~ 8 DEG C, magnetic nanoparticle not sedimentation completely; Under 37 DEG C of environment, there is sedimentation in 3 days in magnetic nanoparticle, within 6 days, do not have complete sedimentation.
Conserving liquid embodiment 2 being dispersed with to magnetic nanoparticle carries out accelerated stability test test: get the conserving liquid that 100 μ L are dispersed with magnetic nanoparticle and add to 1.5mL centrifuge tube, make 40,20 are placed in 2 DEG C ~ 8 DEG C Refrigerator stores, and other 20 are placed in 37 DEG C of baking oven accelerated deteriorations; All the other method of testings are identical with performance test, obtain stability test and the results are shown in Table 4.
Table 4 embodiment 2 stability test result
Embodiment 3
700ml aqua sterilisa is added in 1000ml conical flask, then 2.1gMES, 0.24gNaOH, 2.5gNaCl, 0.9gKCl, 50gBSA, 150g glycerine, 0.3g sodium azide is added and 0.5gProclin300 is ultrasonic mixes, filter, 40 DEG C of heating water baths, add 7.5g PVP K30,6g carragheen ι type, 12g Bos taurus domesticus Gmelin and 1g polyvinyl alcohol (PVA) to dissolving completely in batches, then add water and be settled to 1000ml, obtain conserving liquid.
Magnetic nanoparticle NT-BNP being wrapped quilt is dispersed to conserving liquid.
Performance test: accurately pipette 100 μ l, 0.1mol/LPB6.5 to 1.5mL centrifuge tube with liquid-transfering gun, add the conserving liquid that 10 μ l3000ng/mlNT-BNP antigens or 9000ng/mlNT-BNP antigen, the antibody of 20 μ l alkali phosphatase enzyme marks and 100 μ l are dispersed with magnetic nanoparticle successively wherein, then with liquid-transfering gun, mixed liquor is broken up evenly, start timing.After 5min, above-mentioned centrifuge tube is placed in magnetic frame, after 30s, sucks clear liquid, wash twice with 300 μ l0.01mol/LPB7.4.With 100 μ l, 0.2mol/L, pH=9.0Tris-HCl damping fluids, washed magnetic nanoparticle is uniformly dispersed, add 50 μ lCDPStar, mix rear beginning timing, be added to 96 hole white ELISA Plate, with the luminous value of Promega semi-automatic luminometer test 3min.Repeat above-mentioned steps, prepare 3 parallel samples altogether, count No. 1, No. 2 and No. 3, obtain the results are shown in Table 5.
Table 5 embodiment 3Promega luminous value test result
| Promega test value (RLU) | No. 1 | No. 2 | No. 3 | Mean value |
| Blank | 854 | 993 | 769 | 872 |
| NT-BNP antigen 3000ng/mL | 6458 | 7522 | 5937 | 6639 |
| NT-BNP antigen 9000ng/mL | 22356 | 21437 | 23009 | 22267 |
| NT-BNP sample 2459ng/mL | 6278 | 6041 | 6319 | 6212 |
| NT-BNP sample 10792ng/mL | 26790 | 28421 | 24553 | 26588 |
Conserving liquid embodiment 3 being dispersed with to magnetic nanoparticle carries out retention test, obtains result to be: 2 DEG C ~ 8 DEG C, magnetic nanoparticle not sedimentation completely; Under 37 DEG C of environment, there is sedimentation in 3 days in magnetic nanoparticle, within 6 days, do not have complete sedimentation.
Conserving liquid embodiment 3 being dispersed with to magnetic nanoparticle carries out accelerated stability test test: get the conserving liquid that 100 μ L are dispersed with magnetic nanoparticle and add to 1.5mL centrifuge tube, make 40,20 are placed in 2 DEG C ~ 8 DEG C Refrigerator stores, and other 20 are placed in 37 DEG C of baking oven accelerated deteriorations; All the other method of testings are identical with performance test, obtain stability test and the results are shown in Table 4.
Table 6 embodiment 2 stability test result
Claims (10)
1. a conserving liquid, is characterized in that, comprising:
2. conserving liquid according to claim 1, is characterized in that, also comprises the thickening agent of 1 ~ 100g/L except carragheen.
3. conserving liquid according to claim 2, it is characterized in that, described thickening agent except carragheen be selected from gelatin, sodium alginate, hydroxyethyl cellulose, gellan gum, sucrose, polyvinyl alcohol (PVA), polyglycol and polyvinylpyrrolidone one or more.
4. conserving liquid according to claim 1, it is characterized in that, described buffer salt is one or more in sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, 3-(N-morpholinyl) propane sulfonic acid, 2-(N-morpholine) ethyl sulfonic acid monohydrate, NaOH, hydrochloric acid, sodium chloride and potassium chloride.
5. conserving liquid according to claim 1, is characterized in that, described antibody stabilization agent is one or more in casein, bovine serum albumin(BSA), cow's serum and glycerine.
6. conserving liquid according to claim 1, is characterized in that, described antiseptic and inhibiting bacteria function agent is one or more in sodium azide, Proclin300 and chloromycetin.
7. conserving liquid according to claim 1, is characterized in that, described carragheen is one or more in κ type carragheen, ι type carragheen and λ type carragheen.
8. a preparation method for conserving liquid, is characterized in that, comprising:
0.1 ~ 10g/L buffer salt, the agent of 100 ~ 300g/L antibody stabilization, the agent of 0.1 ~ 1g/L antiseptic and inhibiting bacteria function, 2 ~ 12g/L carragheen are mixed with water, heating for dissolving, obtains conserving liquid.
9. preparation method according to claim 8, is characterized in that, the temperature of described heating is 35 DEG C ~ 85 DEG C.
10. the conserving liquid described in claim 1 ~ 7 any one or the application of the conserving liquid prepared by claim 8 ~ 9 any one in magnetic particle or magnetic nanoparticle are preserved.
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| CN111812313B (en) * | 2020-06-22 | 2021-10-08 | 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 |
| 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 |
| 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 |
| CN114384239A (en) * | 2021-12-09 | 2022-04-22 | 深圳君和生物科技有限公司 | Immunomagnetic bead storage system and immunomagnetic bead system |
| CN114397443A (en) * | 2021-12-09 | 2022-04-26 | 华南理工大学 | Immunomagnetic bead preservation solution and immunomagnetic bead reagent |
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