CN102375056A - Immobilized bio-macromolecular stabilizer as well as preparation method and application thereof - Google Patents
Immobilized bio-macromolecular stabilizer as well as preparation method and application thereof Download PDFInfo
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- CN102375056A CN102375056A CN2010102646689A CN201010264668A CN102375056A CN 102375056 A CN102375056 A CN 102375056A CN 2010102646689 A CN2010102646689 A CN 2010102646689A CN 201010264668 A CN201010264668 A CN 201010264668A CN 102375056 A CN102375056 A CN 102375056A
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Abstract
The invention discloses an immobilized bio-macromolecular stabilizer as well as a preparation method and application thereof. The stabilizer comprises the following components: glycerol, amino acids, carbohydrate, peptone, ion chelants and preservatives. The preparation method comprises the following steps of: mixing and dissolving the components, and filtering and sterilizing the mixture to obtain the stabilizer disclosed by the invention. The stabilizer disclosed by the invention has the advantages of low production cost, obvious effect on protection of bio-macromolecular activity and long service life; and the stabilizer can be used for preparing an ELISA (Enzyme-Linked Immuno Sorbent Assay) plate stabilizer with high quality and low price, thereby improving the coating activity of an ELISA.
Description
Technical field
The present invention relates to a kind of stabilizing agent; Relate in particular to macromolecular stabilizing agent of a kind of fixed biologically and preparation method thereof; The invention still further relates to this stabilizing agent in the purposes that improves ELISA ELISA Plate coated antibody or the stability of antigen in the substrate surface dry run, belong to the stabilizing agent field of fixing biological molecules.
Background technology
In some immunological experiment programs, biomolecule need at first be fixed to substrate surface, and for example microwell plate is managed, latex particle, and nitrocellulose filter, pottery, silicon etc., after super-dry, fixing molecular biological activity will weaken even lose then.It is exactly that the kit antibody or the antigen immune activity that in storage life, guarantee to be coated on the ELISA Plate do not change that the ELISA kit carries out one of key problem in technology of commodity production; Be no lack of both at home and abroad at present various commercial diagnostic kit productions and application are arranged, but to the report of stabilizing agent seldom.
Summary of the invention
One of the object of the invention provides a kind of stabilizing agent of fixing biological molecules, can be used for improving ELI SA ELISA Plate coated antibody or the stability of antigen in the substrate surface dry run, improves the shelf life of test kit, improves its commercial value.
Two of the object of the invention provides a kind of method for preparing the stabilizing agent of said fixing biomolecule.
Above-mentioned purpose of the present invention realizes through following technical scheme:
A kind of stabilizing agent of fixing biological molecules comprises following component: glycerine, amino acids, glucide, peptone, ion chelating agent and antiseptic;
Preferably, the consumption of each component is: glycerine 0%～10% (V/V), amino acid 0%～5.0% (m/v); Glucide 0%～10.0% (m/v), peptone 1.0%～8.0% (m/v), ion chelating agent 0.01%～5.0% (m/v); Antiseptic 0.001%～2.0% (m/v), surplus are water.
Preferred; The consumption of each component is: glycerine 0.5%～10% (V/V), amino acid 0.05%～5.0% (m/v), glucide 5.0%～10.0% (m/v); Peptone 1.0%～5.0% (m/v); Ion chelating agent 0.01%～3.0% (m/v), antiseptic 0.001%～1.0% (m/v), surplus is a water.
Preferred especially, the consumption of each component is: glycerine 0.5% (v/v), and amino acid 0.05% (m/v), peptone 1.0% (m/v), ion chelating agent 0.01% (m/v), antiseptic 0.001% (m/v), surplus is a water.
Why select for use amino acid to be in the stabilizing agent of the present invention because it has certain antioxidation, said amino acids includes but not limited to: amino acid such as glycocoll, alanine, arginine, lysine or glutamine.
Said glucide includes but not limited to glucides such as trehalose, sucrose, D-sorbite, sweet mellow wine or lactose.
Said ion chelating agent includes but not limited to EDTA, EDTA-Na
2, EDTA-Na
4Deng material.Said antiseptic includes but not limited to materials such as Sodium azide, thimerosal.
Another object of the present invention provides a kind of method for preparing the stabilizing agent of said fixing biomolecule, and this method may further comprise the steps:
(1) measures each component by said usefulness;
(2) with each component mixed dissolution, filtration sterilization promptly gets.
Stabilizing agent production cost of the present invention is low, the protection effect is obvious, the effect phase is long, can be used for preparing the ELISA Plate stabilizing agent of super quality and competitive price, improves ELISA and encapsulates activity, has higher commercial value.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment with form or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall in protection scope of the present invention the details of technical scheme of the present invention.
The preparation of embodiment 1 stabilizing agent
(1) measure each component by following usefulness: glycerine 1.0% (v/v), glycocoll 1.0% (m/v), trehalose 5.0% (m/v), peptone 2.0% (m/v), ion chelating agent EDTA1.0% (m/v), antiseptic Sodium azide 0.5% (m/v), surplus is a water;
(2) with above-mentioned each component mixed dissolution, filtration sterilization promptly gets.
The preparation of embodiment 2 stabilizing agents
(1) measures each component by following usefulness: glycerine 0.5% (v/v), alanine 0.05% (m/v), peptone 1.0% (m/v), ion chelating agent EDTA-Na
20.01% (m/v), antiseptic Sodium azide 0.001% (m/v), surplus is a water;
(2) with above-mentioned each component mixed dissolution, filtration sterilization promptly gets.
The preparation of embodiment 3 stabilizing agents
(1) measure each component by following usefulness: glycerine 10.0% (v/v), lysine 5.0% (m/v), trehalose 10.0% (m/v), peptone 5.0% (m/v), ion chelating agent 3.0% (m/v), antiseptic Sodium azide 1.0% (m/v), surplus is a water;
(2) with above-mentioned each component mixed dissolution, filtration sterilization promptly gets.
The preparation of embodiment 4 stabilizing agents
(1) measures each component by following usefulness: glycerine 10% (v/v), glutamine 0.05% (m/v), peptone 5.0% (m/v), ion chelating agent EDTA-Na
23.0% (m/v), antiseptic Sodium azide 0.001% (m/v), surplus is a water;
(2) with above-mentioned each component mixed dissolution, filtration sterilization promptly gets.
The preparation of embodiment 5 stabilizing agents
(1) measures each component by following usefulness: glycocoll 5.0% (m/v), trehalose 5.0% (m/v), peptone 5.0% (m/v), ion chelating agent EDTA-Na
43.0% (m/v), antiseptic thimerosal 1.0% (m/v), surplus is a water;
(2) with above-mentioned each component mixed dissolution, filtration sterilization promptly gets.
Test Example 1 stabilizing agent of the present invention is improving the effect test of biomolecule stability
Supply test agent: the stabilizing agent that embodiment 1-5 is prepared is numbered 1-5 respectively;
Test method: 5 kinds of stabilizing agents difference pack processing using preparation are by the ELISA Plate of last antibody or antigen; If place 3d, 6d, 12d for control group places in 37 ℃ of constant temperature ovens without what stabilizing agent was handled; Balance 30min under the room temperature detects the OD value with double antibodies sandwich ELISA method, and test findings is seen table 1.
Table 1 different stabilizers is handled and is encapsulated the OD value after 37 ℃ of held of plate without what stabilizing agent was handled
Generally speaking, antibody or antigen were equivalent to shelf life 45 days 37 ℃ of following storage at a constant temperature 1 day.Test findings shows in the table 1; Handle the coated elisa plate activity through different stabilizers variation is in various degree all arranged; Without 37 ℃ of following storage at a constant temperature of stabilizing agent treatment enzyme target 3 days, activity had largely and reduces, lower temperature preserve encapsulate plate active 23.14%; 37 ℃ of following storage at a constant temperature 12 days, activity is lost basically.With stabilizing agent 1-5 number coated elisa plate is handled back 37 ℃ respectively and quicken storage test, the result shows in the table, and activity compared under the no stabilizing agent situation raising is in various degree all arranged after the 1-5 stabilizing agent was handled coated elisa plate; Especially No. 2 stabilizing agents, effect is more obvious, 37 ℃ of following storage at a constant temperature 6 days; Activity still remains on 88.5%; 37 ℃ of following storage at a constant temperature 12 days, ELISA Plate is active to detect 51.8%, and the coated elisa plate shelf life can be near 1 year; No. 2 stabilizing agent has stablizing effect preferably to coated elisa plate, in ELISA kit developing, development, has bigger application potential.
Claims (10)
1. the macromolecular stabilizing agent of fixed biologically comprises following component: glycerine, amino acids, glucide, peptone, ion chelating agent and antiseptic.
2. according to the described stabilizing agent of claim 1, it is characterized in that the consumption of each component is: by V/V, glycerine 0%～10%; By m/v: amino acid 0%～5.0%, glucide 0%～10.0%, peptone 1.0%～8.0%, ion chelating agent 0.01%～5.0%, antiseptic 0.001%～2.0%; Surplus is a water.
3. according to the described stabilizing agent of claim 2, it is characterized in that the consumption of each component is: by V/V, glycerine 0.5%～10%; By m/v: amino acid 0.05%～5.0%, glucide 5.0%～10.0%, peptone 1.0%～5.0%, ion chelating agent 0.01%～3.0%, antiseptic 0.001%～1.0%; Surplus is a water.
4. according to the described stabilizing agent of claim 2, it is characterized in that the consumption of each component is: by V/V: glycerine 0.5% (v/v); By m/v: amino acid 0.05%, peptone 1.0%, ion chelating agent 0.01%, antiseptic 0.001%; Surplus is a water.
5. according to the described stabilizing agent of claim 1, it is characterized in that: said amino acids includes but not limited to: glycocoll, alanine, arginine, lysine or glutamine.
6. according to the described stabilizing agent of claim 1, it is characterized in that: said glucide includes but not limited to trehalose, sucrose, D-sorbite, sweet mellow wine or lactose.
7. according to the described stabilizing agent of claim 1, it is characterized in that: said ion chelating agent includes but not limited to EDTA, EDTA-Na
2Or EDTA-Na
4Said antiseptic includes but not limited to Sodium azide or thimerosal.
8. method for preparing any one said stabilizing agent of claim 1-7, may further comprise the steps: each component is measured by said usefulness in (1); (2) with each component mixed dissolution, filtration sterilization promptly gets.
9. any one said stabilizing agent of claim 1-7 is fixed in the purposes in the stability of biomacromolecule of substrate surface in raising.
10. according to the described purposes of claim claim 9, it is characterized in that: described biomacromolecule comprises protein, protein derivatives or DNA.
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| CN2010102646689A CN102375056A (en) | 2010-08-27 | 2010-08-27 | Immobilized bio-macromolecular stabilizer as well as preparation method and application thereof |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103048448A (en) * | 2012-12-21 | 2013-04-17 | 杭州茂天赛科技有限公司 | Stationary liquid |
| CN103472222A (en) * | 2013-08-26 | 2013-12-25 | 河北省科学院生物研究所 | Long-acting ELISA plate stabilizing agent |
| CN103626830A (en) * | 2013-05-21 | 2014-03-12 | 郑州牧业工程高等专科学校 | Normal-temperature storage liquid of transfer factors for livestock and poultry |
| CN104459147A (en) * | 2013-09-18 | 2015-03-25 | 深圳迈瑞生物医疗电子股份有限公司 | Biomarker preserving liquid, biomarker reagent and method |
| CN105018457A (en) * | 2015-07-20 | 2015-11-04 | 宁波美成生物科技有限公司 | Luciferase stabilizer |
| CN107656045A (en) * | 2017-11-01 | 2018-02-02 | 上海凯创生物技术有限公司 | Procalcitonin near-infrared fluorescent detection reagent card, kit and application thereof |
| CN107884584A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Rotavirus detection reagent card, kit and application thereof |
| CN107884575A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Amino terminal-pro brain natriuretic peptide detection reagent card, kit and application thereof |
| CN107884574A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Chlamydia trachomatis detection reagent card, kit and application thereof |
| CN107907681A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | Adenovirus detection reagent card, kit and application thereof |
| CN107907678A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | D dimer quantum dot fluorescence detection reagents card, kit and application thereof |
| CN107907514A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | CTnI/myoglobins/creatine kinase isozyme near-infrared fluorescent detection reagent card, kit and application thereof |
| CN107941742A (en) * | 2017-11-01 | 2018-04-20 | 上海凯创生物技术有限公司 | β human chorionic gonadotrophin near-infrared fluorescent detection reagents card, kit and application thereof |
| CN109765385A (en) * | 2019-01-29 | 2019-05-17 | 浙江夸克生物科技有限公司 | A method of improving emulsion reagent sensitivity and linear |
| CN111122879A (en) * | 2020-03-26 | 2020-05-08 | 珠海丽珠试剂股份有限公司 | Coating liquid of novel coronavirus recombinant antigen, pretreatment method, application and product |
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Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103048448A (en) * | 2012-12-21 | 2013-04-17 | 杭州茂天赛科技有限公司 | Stationary liquid |
| CN103626830A (en) * | 2013-05-21 | 2014-03-12 | 郑州牧业工程高等专科学校 | Normal-temperature storage liquid of transfer factors for livestock and poultry |
| CN103626830B (en) * | 2013-05-21 | 2015-07-15 | 郑州牧业工程高等专科学校 | Normal-temperature storage liquid of transfer factors for livestock and poultry |
| CN103472222A (en) * | 2013-08-26 | 2013-12-25 | 河北省科学院生物研究所 | Long-acting ELISA plate stabilizing agent |
| CN103472222B (en) * | 2013-08-26 | 2015-12-23 | 河北省科学院生物研究所 | A kind of long-acting ELISA Plate stabilizing agent |
| CN104459147A (en) * | 2013-09-18 | 2015-03-25 | 深圳迈瑞生物医疗电子股份有限公司 | Biomarker preserving liquid, biomarker reagent and method |
| CN105018457A (en) * | 2015-07-20 | 2015-11-04 | 宁波美成生物科技有限公司 | Luciferase stabilizer |
| CN107884584A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Rotavirus detection reagent card, kit and application thereof |
| CN107656045A (en) * | 2017-11-01 | 2018-02-02 | 上海凯创生物技术有限公司 | Procalcitonin near-infrared fluorescent detection reagent card, kit and application thereof |
| CN107884575A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Amino terminal-pro brain natriuretic peptide detection reagent card, kit and application thereof |
| CN107884574A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Chlamydia trachomatis detection reagent card, kit and application thereof |
| CN107907681A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | Adenovirus detection reagent card, kit and application thereof |
| CN107907678A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | D dimer quantum dot fluorescence detection reagents card, kit and application thereof |
| CN107907514A (en) * | 2017-11-01 | 2018-04-13 | 上海凯创生物技术有限公司 | CTnI/myoglobins/creatine kinase isozyme near-infrared fluorescent detection reagent card, kit and application thereof |
| CN107941742A (en) * | 2017-11-01 | 2018-04-20 | 上海凯创生物技术有限公司 | β human chorionic gonadotrophin near-infrared fluorescent detection reagents card, kit and application thereof |
| CN109765385A (en) * | 2019-01-29 | 2019-05-17 | 浙江夸克生物科技有限公司 | A method of improving emulsion reagent sensitivity and linear |
| CN111122879A (en) * | 2020-03-26 | 2020-05-08 | 珠海丽珠试剂股份有限公司 | Coating liquid of novel coronavirus recombinant antigen, pretreatment method, application and product |
| CN111122879B (en) * | 2020-03-26 | 2020-07-14 | 珠海丽珠试剂股份有限公司 | Coating liquid of novel coronavirus recombinant antigen, pretreatment method, application and product |
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Application publication date: 20120314 |