CN115340910A - Cleaning solution suitable for alkaline phosphatase chemiluminescence system - Google Patents
Cleaning solution suitable for alkaline phosphatase chemiluminescence system Download PDFInfo
- Publication number
- CN115340910A CN115340910A CN202211084813.4A CN202211084813A CN115340910A CN 115340910 A CN115340910 A CN 115340910A CN 202211084813 A CN202211084813 A CN 202211084813A CN 115340910 A CN115340910 A CN 115340910A
- Authority
- CN
- China
- Prior art keywords
- cleaning solution
- alkaline phosphatase
- surfactant
- cleaning
- comparative example
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000004140 cleaning Methods 0.000 title claims abstract description 67
- 102000002260 Alkaline Phosphatase Human genes 0.000 title claims abstract description 29
- 108020004774 Alkaline Phosphatase Proteins 0.000 title claims abstract description 29
- 239000000243 solution Substances 0.000 claims abstract description 64
- 239000004094 surface-active agent Substances 0.000 claims abstract description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 13
- 239000002736 nonionic surfactant Substances 0.000 claims abstract description 9
- 239000007983 Tris buffer Substances 0.000 claims abstract description 7
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000002518 antifoaming agent Substances 0.000 claims description 11
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 8
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 8
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 8
- 230000002335 preservative effect Effects 0.000 claims description 8
- 239000003755 preservative agent Substances 0.000 claims description 7
- 239000004721 Polyphenylene oxide Substances 0.000 claims description 5
- 229920000570 polyether Polymers 0.000 claims description 5
- 238000003018 immunoassay Methods 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 14
- 238000012360 testing method Methods 0.000 abstract description 11
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 230000003197 catalytic effect Effects 0.000 abstract description 3
- 229910001425 magnesium ion Inorganic materials 0.000 abstract description 2
- 230000002195 synergetic effect Effects 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 29
- 239000007788 liquid Substances 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 238000004020 luminiscence type Methods 0.000 description 5
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 5
- 229920002545 silicone oil Polymers 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 239000011324 bead Substances 0.000 description 3
- 230000027455 binding Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 3
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- UMCMPZBLKLEWAF-BCTGSCMUSA-N 3-[(3-cholamidopropyl)dimethylammonio]propane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 UMCMPZBLKLEWAF-BCTGSCMUSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- XYIPYISRNJUPBA-UHFFFAOYSA-N [3-(3'-methoxyspiro[adamantane-2,4'-dioxetane]-3'-yl)phenyl] dihydrogen phosphate Chemical compound O1OC2(C3CC4CC(C3)CC2C4)C1(OC)C1=CC=CC(OP(O)(O)=O)=C1 XYIPYISRNJUPBA-UHFFFAOYSA-N 0.000 description 2
- 239000013530 defoamer Substances 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229960003415 propylparaben Drugs 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920002366 Tetronic® 1307 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- XVBRCOKDZVQYAY-UHFFFAOYSA-N bronidox Chemical compound [O-][N+](=O)C1(Br)COCOC1 XVBRCOKDZVQYAY-UHFFFAOYSA-N 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 239000003093 cationic surfactant Substances 0.000 description 1
- 150000001768 cations Chemical group 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- KAKKHKRHCKCAGH-UHFFFAOYSA-L disodium;(4-nitrophenyl) phosphate;hexahydrate Chemical compound O.O.O.O.O.O.[Na+].[Na+].[O-][N+](=O)C1=CC=C(OP([O-])([O-])=O)C=C1 KAKKHKRHCKCAGH-UHFFFAOYSA-L 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000036046 immunoreaction Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 229910052816 inorganic phosphate Inorganic materials 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000004712 monophosphates Chemical class 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- UEUXEKPTXMALOB-UHFFFAOYSA-J tetrasodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical class [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O UEUXEKPTXMALOB-UHFFFAOYSA-J 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D1/00—Detergent compositions based essentially on surface-active compounds; Use of these compounds as a detergent
- C11D1/66—Non-ionic compounds
- C11D1/835—Mixtures of non-ionic with cationic compounds
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/02—Inorganic compounds ; Elemental compounds
- C11D3/04—Water-soluble compounds
- C11D3/046—Salts
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/26—Organic compounds containing nitrogen
- C11D3/30—Amines; Substituted amines ; Quaternized amines
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/37—Polymers
- C11D3/3703—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
- C11D3/3707—Polyethers, e.g. polyalkyleneoxides
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D1/00—Detergent compositions based essentially on surface-active compounds; Use of these compounds as a detergent
- C11D1/38—Cationic compounds
- C11D1/62—Quaternary ammonium compounds
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D1/00—Detergent compositions based essentially on surface-active compounds; Use of these compounds as a detergent
- C11D1/66—Non-ionic compounds
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D1/00—Detergent compositions based essentially on surface-active compounds; Use of these compounds as a detergent
- C11D1/66—Non-ionic compounds
- C11D1/74—Carboxylates or sulfonates esters of polyoxyalkylene glycols
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
- C11D2111/10—Objects to be cleaned
- C11D2111/14—Hard surfaces
- C11D2111/20—Industrial or commercial equipment, e.g. reactors, tubes or engines
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Inorganic Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Detergent Compositions (AREA)
Abstract
The invention discloses a cleaning solution suitable for an alkaline phosphatase chemiluminescence system, wherein the pH of the cleaning solution is 9-10, and the cleaning solution specifically comprises Tris, naCl and MgCl 2 The surfactant comprises a nonionic surfactant and a Gemini surfactant. The cleaning capacity of the cleaning solution is enhanced and the variation coefficient in the test is reduced through the high pH and the synergistic effect of the nonionic surfactant and the Gemini surfactant; moreover, the catalytic site of alkaline phosphatase is activated in advance by the coordination of Gemini surfactant and Mg ions, which is beneficial toThe detection signal value is improved, and the detection sensitivity is improved.
Description
Technical Field
The invention belongs to the technical field of chemiluminescence, and particularly relates to a cleaning solution suitable for an alkaline phosphatase chemiluminescence system.
Background
The full-automatic chemiluminescence determinator combines a high-sensitivity chemiluminescence determining technology with high-specificity immunoreaction, has the advantages of simplicity and convenience in use and operation, reliable result, stable performance and the like, and becomes one of the instruments with the highest use frequency in clinical laboratory departments in hospitals. The alkaline phosphatase chemiluminescence system is one of the systems commonly used in chemiluminescence measuring instruments, and the alkaline phosphatase can hydrolyze various monophosphates and release inorganic phosphate. In an alkaline phosphatase system, a1, 2-dioxetane compound (AMPPD) is used as an ultra-sensitive alkaline phosphatase substrate, under the catalytic action of alkaline phosphatase, a phosphate group of the AMPPD is hydrolyzed to form an unstable intermediate, and after the intermediate is decomposed by self, a four-membered ring is broken to release a large amount of energy, so that a chemiluminescence reaction is excited.
The cleaning solution is used as a general reagent of a full-automatic chemiluminescence determinator, is often matched with various immunoassay kits for use, and is used for cleaning substances which do not participate in specific reaction in a reaction cup and on a needle head, so that abnormal fluctuation of a detection result is avoided, and interference caused by non-specific binding is reduced. The good cleaning solution can clean the cup wall, the needle, the magnetic bead and the substances with non-specific and weak combination of the antibody and the antigen to the maximum extent on the basis of not influencing the specific combination of the antigen and the antibody and the function of the alkaline phosphatase so as to improve the detection sensitivity and precision and reduce the detection variation coefficient.
CN111607465A discloses a chemiluminescent cleaning solution, a preparation method and an application thereof, which mainly comprises Tris, sodium ions, CHAPS, triton-100, PC300, BND, propylparaben and silicone oil type defoaming agent, and the pH value of the cleaning solution is 7.0-7.5.CN110862881A discloses a cleaning solution or a diluent thereof special for a full-automatic chemiluminescence determinator and a preparation method thereof, and the cleaning solution or the diluent mainly comprises inorganic salt, a phosphate buffer system, a surfactant 1 (Tween-20 or Tween 80), a surfactant 2 (sodium dodecyl sulfate or sodium monthly silicate), a preservative (Proclin 300 and Bronidox), and the pH value is 7.2-8.3.CN113484306A discloses a magnetic particle chemiluminescence cleaning solution and a preparation method thereof, which mainly comprises a phosphate buffer system, inorganic salt, EDTA sodium salt, tween 20, sodium dodecyl sulfate, PC300, sodium azide, a silicone oil type defoaming agent, and the pH value is 7.0-7.5. In the above patent application, there are the following problems: (1) phosphate buffer systems are not suitable for alkaline phosphatase chemiluminescence systems because phosphate inhibits alkaline phosphatase, resulting in a decrease in detection signal and thus insufficient detection sensitivity for some items; (2) the sodium salt of EDTA will chelate the active metal ions Mg and Zn of alkaline phosphatase, resulting in the inactivation of alkaline phosphatase; (3) although the silicone oil type antifoaming agent can effectively suppress foaming, it affects antigen-antibody binding, and the silicone oil type antifoaming agent carried into the reagent in a small amount also seriously affects the stability of the reagent; (4) propyl p-hydroxybenzoate, sodium azide and the like are flammable or explosive substances, and the safety is low.
Disclosure of Invention
In view of the above, the present invention provides a cleaning solution suitable for an alkaline phosphatase chemiluminescence system, which can improve the cleaning ability and further improve the detection sensitivity.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
the cleaning liquid suitable for alkaline phosphatase chemiluminescence system has a pH of 9-10, and specifically comprises Tris, naCl, mgCl 2 Surfactants and defoamers; wherein the surfactant is composed of a nonionic surfactant and a Gemini surfactant.
Further, in the above technical solution, the defoaming agent is a polyether type defoaming agent, and the addition amount is 0.001 to 0.1wt%.
Further, in the above technical scheme, the cleaning also comprises a preservative with an addition amount of 0.1-2 wt%; still further, the preservative is PC-950 and/or BND-10.
Furthermore, in the technical scheme, the addition amount of the Gemini surfactant is 0.001-0.1 wt%. The Gemini surfactant is formed by connecting the heads of two same cationic surfactants through hydrophobic groups, and has more outstanding characteristic properties due to the unique double hydrophilic head structure, such as easier adsorption on gas/liquid surfaces, more effective reduction of the surface tension of aqueous solutions and easier aggregation to generate micelles.
Specifically, the anion of the Gemini surfactant is Cl ion or Br ion, and the cation structure of the Gemini surfactant is as follows:
wherein n has a value of 6 to 20 and m has a value of 2 to 6 in the cationic structure.
Further, in the above technical solution, the non-ionic surfactant comprises tween 20 and S9 (ethylenediamine alkoxylate block copolymer, also called Tetronic 1307), the addition amount of tween 20 is 0.1-2 wt%, and the addition amount of S9 is 0.1-2 wt%.
Further, in the above technical solution, the amount of NaCl added is 1-10 wt%.
Further, in the above technical scheme, mgCl 2 The addition amount of (B) is 1-10 wt%.
The invention further provides a preparation method of the cleaning solution, which comprises the following steps: taking pure water as a solvent, and adding Tris, naCl and MgCl in proportion 2 The water-soluble organic silicon dioxide gel is prepared by uniformly stirring a nonionic surfactant, a preservative, a Gemini surfactant and a polyether defoaming agent, adjusting the pH value to 9-10 by using hydrochloric acid, and finally filtering the solution by using a 0.22-micron filter membrane.
Because the using amount of the cleaning solution is large, in order to save the transportation cost, the cleaning solution prepared by the invention is concentrated, and when the cleaning solution provided by the invention is used for the cleaning step of chemiluminescence immunoassay, pure water is firstly used for diluting by 10-20 times.
Compared with the prior art, the invention has the beneficial effects that:
1) The pH value of the cleaning solution provided by the invention is 9-10, the high pH value is beneficial to activating alkaline phosphatase on one hand, and the nonspecific combination of antigen and antibody can be eluted to the maximum extent on the other hand, and experiments prove that the cleaning solution causes dissociation between the antigen and the antibody under the pH condition, thereby causing the loss of effective signals; in addition, the PC-950 and BND-10 are used as preservatives under the pH condition to have better preservative effect, and the PC-300 loses the preservative property due to yellowing;
2) The invention discloses a method for cleaning a magnetic bead or a test tube by using a Gemini surfactant, which comprises the following steps of (1) simultaneously adopting a nonionic surfactant and the Gemini surfactant in a cleaning solution, finding that the non-specifically bound hybrid protein can be cleaned off under the condition of not damaging the binding of an antigen and an antibody by using a very small amount of the Gemini surfactant, reducing the physical adsorption between the protein and the wall of the cup and a needle head by using the nonionic surfactants Tween 20 and S9, reducing the alkaline phosphatase non-specifically bound on the magnetic bead or the test tube by the synergistic effect of the protein and the wall of the cup and the needle head, and further reducing the variation coefficient during testing;
3) In the present invention, a large amount of inorganic salts (i.e., naCl and MgCl) are added 2 ) So as to provide electrolyte suitable for immune reaction and reduce the binding capacity of non-target protein and antibody; in addition, the Gemini surfactant is matched with Mg ions, so that the catalytic sites of alkaline phosphatase can be quickly activated in advance, and the detection sensitivity is improved;
4) The invention adopts the polyether type defoaming agent, which not only can effectively prevent bubbles from being generated in the washing process, but also can not influence the stability of the detection reagent and avoid the jump value phenomenon.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are given by way of illustration and explanation only, not limitation.
In the following examples, unless otherwise specified, all the methods are conventional; the reagents and materials are commercially available unless otherwise specified.
Example 1
The cleaning liquid provided by the embodiment comprises: 2% Tris, 10% NaCl, 5% MgCl 2 2% tween 20, 1% s9, 1% pc-950, 1% bnd-10, 0.1% gemini surfactant (in particular n =12, m =4 chloride) and 0.1% polyether defoamer (defoamer 204 of Sigma). Adding the raw materials into pure water according to a certain proportion, stirring until the mixture is uniformly mixed, adjusting the pH value to 9.5 by using hydrochloric acid, and filtering the solution by using a 0.22 mu m filter membrane.
The cleaning liquid provided in this example is preferably diluted 20 times with pure water at the time of use.
Comparative example 1
The cleaning solution provided in this example does not contain a Gemini surfactant, and the rest is the same as example 1.
Comparative example 2
This example provides a cleaning solution that does not contain MgCl 2 Otherwise, the same procedure as in example 1 was repeated.
Comparative example 3
The cleaning solution provided by the embodiment is an optimal formula of Chinese patent CN111607465A, and specifically comprises the following components: 500mM Tris buffer, 0.5% CHAPS, 0.5% Triton X-100, 500mM NaCl, 0.6% Proclin-300, 0.1% BND, 0.1% propylparaben, 0.1% silicone oil type antifoam, and pH 7.3.
The cleaning liquids in example 1, comparative example 1 and comparative example 2 were colorless transparent liquids in appearance, and the cleaning liquid in comparative example 3 was a colorless transparent liquid having oily floats.
The influence of the cleaning solution of example 1, comparative example 2 and comparative example 3 on the detection result is further detected by a full-automatic chemiluminescence determinator (crude source SZY-CL1000 instrument) and a CA125 luminescence kit (crude source research reagent) and a G17 luminescence kit (crude source research reagent), and specifically comprises the following aspects:
(1) And (5) detecting a blank calibrator.
The zero concentration calibrator of each kit was repeatedly measured 20 times using the wash solutions example 1, comparative example 2, and comparative example 3, respectively, according to the instrument operation flow and the luminescence kit instructions, and the results are shown in the following table (coefficient of variation = (standard deviation SD/Mean) × 100%):
as can be seen from the above table, the cleaning solution of example 1 has a lower blank value than the cleaning solutions of comparative examples 1 to 3, and the coefficient of variation of the cleaning solution of example 1 is also significantly lower than the cleaning solutions of comparative examples 1 to 3.
(2) And (5) detecting a low-value calibrator.
The 10U/ml calibrator of the CA125 luminescence kit and the 5pmol/L calibrator of the G17 luminescence kit were each tested 20 times in duplicate using wash solutions example 1, comparative example 2, and comparative example 3, respectively, and the results are shown in the following table:
as can be seen from the table above, for the same calibrator, the cleaning solution of comparative example 3 not only has a high coefficient of variation, but also has a jump value problem, the coefficient of variation of the cleaning solution of example 1 is less than 5%, and the test signal is significantly higher than that of the cleaning solution of comparative example; although the coefficient of variation of comparative example 1 and comparative example 2 was greatly reduced, the intensity of the test signal was significantly lower than that of example 1.
(3) And (4) detecting the mixed sample.
The pooled samples (randomly pooled clinical serum samples from 10 different persons) were tested 20 times in duplicate using wash solution example 1, comparative example 2 and comparative example 3, respectively, using the CA125 and G17 kits, and the results are shown in the small table:
from the table, it can be seen that, when the same sample is tested, the cleaning solution of comparative example 3 not only has a high coefficient of variation, but also has a jump value problem, the coefficient of variation of the cleaning solution of example 1 is less than 5%, and the test signal is significantly higher than that of the cleaning solution of comparative example. Although the coefficient of variation of comparative examples 1 and 2 was greatly reduced, the test signal intensity and coefficient of variation were significantly inferior to those of example 1.
(4) And (5) detecting the stability of the cleaning solution.
The cleaning solutions of example 1, comparative example 2 and comparative example 3 were left to stand at normal temperature in the dark for 6 months. In appearance, the appearance of the cleaning solution in example 1, comparative example 1 and comparative example 2 is unchanged, and the color of the cleaning solution in comparative example 3 is slightly yellow; the mixed sample was further tested 20 times repeatedly with the CA125 and G17 kits, and the test results are shown in the following table:
as can be seen from the above table, the test variation coefficient of the cleaning solution of example 1 is not significantly changed compared with the sample before 6 months, and the frequency of occurrence of the jump value of the cleaning solution of the comparative example is increased.
The raw materials listed in the invention, and the upper and lower limits and interval values of the raw materials can realize the invention, and the examples are not listed.
In conclusion, the cleaning solution provided by the invention has good stability at normal temperature and in dark place, has strong cleaning capability, has lower blank detection value and test variation coefficient compared with the prior art, can pre-activate alkaline phosphatase during cleaning, improves the detection signal value, and further improves the detection sensitivity of the kit.
The above description is of the preferred embodiment of the present invention and should not be taken as limiting the scope of the invention, but rather, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. The cleaning solution suitable for the alkaline phosphatase chemiluminescence system is characterized in that the pH of the cleaning solution is 9-10, and the cleaning solution comprises Tris, naCl and MgCl 2 The surfactant comprises a nonionic surfactant and a Gemini surfactant.
2. The cleaning solution for alkaline phosphatase chemiluminescent system according to claim 1, wherein the defoaming agent is polyether type defoaming agent and is added in an amount of 0.001-0.1 wt%.
3. The cleaning solution suitable for use in an alkaline phosphatase chemiluminescent system according to claim 1, wherein the cleaning solution further comprises 0.1-2 wt% of a preservative.
4. The cleaning solution suitable for use in a chemiluminescent system of alkaline phosphatase according to claim 3 wherein the preservative is PC-950 and/or BND-10.
5. The cleaning solution suitable for use in alkaline phosphatase chemiluminescent system according to claim 1 wherein the Gemini surfactant is added in an amount of 0.001-0.1 wt%.
6. The cleaning solution suitable for use in alkaline phosphatase chemiluminescence system according to claim 1, wherein the nonionic surfactant comprises tween 20 and S9, the amount of tween 20 added is 0.1 to 2%, and the amount of S9 added is 0.1 to 2%.
7. The cleaning solution for alkaline phosphatase chemiluminescent system according to claim 1, wherein the amount of NaCl added is 1-10 wt%.
8. The cleaning solution for alkaline phosphatase chemiluminescent system according to claim 1, wherein the MgCl is used 2 The addition amount of (B) is 1-10 wt%.
9. Use of a wash solution according to any of claims 1 to 8 in a chemiluminescent immunoassay.
10. The use according to claim 9, wherein the washing solution is diluted 10 to 20 times and used in the washing step of the chemiluminescent immunoassay.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211084813.4A CN115340910B (en) | 2022-09-06 | 2022-09-06 | Cleaning solution suitable for alkaline phosphatase chemiluminescence system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211084813.4A CN115340910B (en) | 2022-09-06 | 2022-09-06 | Cleaning solution suitable for alkaline phosphatase chemiluminescence system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115340910A true CN115340910A (en) | 2022-11-15 |
| CN115340910B CN115340910B (en) | 2023-05-05 |
Family
ID=83955747
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211084813.4A Active CN115340910B (en) | 2022-09-06 | 2022-09-06 | Cleaning solution suitable for alkaline phosphatase chemiluminescence system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115340910B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115873664A (en) * | 2022-12-06 | 2023-03-31 | 上海领骏生物科技有限公司 | Cleaning solution for nucleic acid extraction magnetic beads and use method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013134109A2 (en) * | 2012-03-04 | 2013-09-12 | Traynor Daniel H | Soap with dispersed articles producing light and/or sound |
| CN110408478A (en) * | 2019-07-10 | 2019-11-05 | 广州奕昕生物科技有限公司 | A kind of general washing lotion of enzymatic Full-automatic chemiluminescence |
| CN110862881A (en) * | 2019-11-29 | 2020-03-06 | 郑州安图生物工程股份有限公司 | Special cleaning solution or diluent for full-automatic chemiluminescence determinator and preparation method thereof |
| CN111607465A (en) * | 2020-06-08 | 2020-09-01 | 珠海丽珠试剂股份有限公司 | Chemiluminescent cleaning solution and preparation method and application thereof |
| CN112067601A (en) * | 2020-08-05 | 2020-12-11 | 武汉生之源生物科技股份有限公司 | Alkaline phosphate enzymatic chemiluminescence substrate reinforcing agent and application thereof |
-
2022
- 2022-09-06 CN CN202211084813.4A patent/CN115340910B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013134109A2 (en) * | 2012-03-04 | 2013-09-12 | Traynor Daniel H | Soap with dispersed articles producing light and/or sound |
| CN110408478A (en) * | 2019-07-10 | 2019-11-05 | 广州奕昕生物科技有限公司 | A kind of general washing lotion of enzymatic Full-automatic chemiluminescence |
| CN110862881A (en) * | 2019-11-29 | 2020-03-06 | 郑州安图生物工程股份有限公司 | Special cleaning solution or diluent for full-automatic chemiluminescence determinator and preparation method thereof |
| CN111607465A (en) * | 2020-06-08 | 2020-09-01 | 珠海丽珠试剂股份有限公司 | Chemiluminescent cleaning solution and preparation method and application thereof |
| CN112067601A (en) * | 2020-08-05 | 2020-12-11 | 武汉生之源生物科技股份有限公司 | Alkaline phosphate enzymatic chemiluminescence substrate reinforcing agent and application thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115873664A (en) * | 2022-12-06 | 2023-03-31 | 上海领骏生物科技有限公司 | Cleaning solution for nucleic acid extraction magnetic beads and use method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115340910B (en) | 2023-05-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Diehl et al. | The versenate titration for total hardness | |
| CN109406796B (en) | Rheumatoid factor detection kit and detection method thereof | |
| CN112129950B (en) | Magnetic particle chemiluminescence kit for detecting interleukin 6 | |
| CN108546602B (en) | Cleaning solution for chemiluminescence immunoassay and magnetic particle chemiluminescence immunoassay detection method | |
| CN115340910A (en) | Cleaning solution suitable for alkaline phosphatase chemiluminescence system | |
| CN111607465B (en) | Chemiluminescent cleaning solution and preparation method and application thereof | |
| JP2500233B2 (en) | Method for measuring calcium and / or magnesium and reagent composition for measurement | |
| CN111896731A (en) | Cleaning solution for chemiluminescence immunoassay | |
| CN111983217A (en) | Sample treatment fluid and application thereof | |
| EP0828159B1 (en) | Method for the detection of lysozyme | |
| CN113136269A (en) | Cleaning solution for chemiluminescence immunoassay | |
| KR20070001937A (en) | Measurement value lowering inhibitor for immunoassay method and immunoassay method using the same | |
| CN113671169A (en) | Non-specific adsorption cleaning solution for chemiluminescence immunoassay AEP | |
| JP6496980B2 (en) | Immunoassay method | |
| CN113484306B (en) | Magnetic particle chemiluminescence cleaning solution and preparation method and application thereof | |
| Edwards et al. | Determination of synthetic detergents in sewage | |
| CN113030458A (en) | Sample diluent for ELISA detection kit and preparation method thereof | |
| CN110865182B (en) | Blocking agent and application thereof in immunodetection | |
| KR101675412B1 (en) | Immunoassay method and reagent therefor | |
| CN104048955A (en) | High density lipoprotein cholesterol detection kit | |
| CN115078711A (en) | Cleaning fluid for flow-type fluorescence detection | |
| Hasebe et al. | Flow injection determination of hydrogen peroxide by bis (2, 4, 6-trichlorophenyl) oxalate chemiluminescence in O/W emulsion | |
| CN116103095A (en) | Magnetic particle chemiluminescent cleaning solution and preparation method thereof | |
| CN110749574B (en) | Method for measuring perfluorooctane sulfonate by dual-wavelength resonance Rayleigh scattering method and application | |
| CN116026666B (en) | Plasma sample diluent for D-dimer magnetic particle enzymatic chemiluminescence kit and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A cleaning solution suitable for alkaline phosphatase chemiluminescence system Granted publication date: 20230505 Pledgee: Guanggu Branch of Wuhan Rural Commercial Bank Co.,Ltd. Pledgor: WUHAN LIFE ORIGIN BIOTECH JOINT STOCK Co.,Ltd. Registration number: Y2024980009814 |