CN110133253A - Broad spectrum type antigen-antibody protective agent - Google Patents
Broad spectrum type antigen-antibody protective agent Download PDFInfo
- Publication number
- CN110133253A CN110133253A CN201910396961.1A CN201910396961A CN110133253A CN 110133253 A CN110133253 A CN 110133253A CN 201910396961 A CN201910396961 A CN 201910396961A CN 110133253 A CN110133253 A CN 110133253A
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- China
- Prior art keywords
- protective agent
- antigen
- antibody
- broad spectrum
- spectrum type
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- 239000003223 protective agent Substances 0.000 title claims abstract description 78
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 24
- 230000000694 effects Effects 0.000 claims abstract description 20
- 238000010790 dilution Methods 0.000 claims abstract description 12
- 239000012895 dilution Substances 0.000 claims abstract description 12
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 7
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 7
- 150000002016 disaccharides Chemical class 0.000 abstract description 12
- 102000036639 antigens Human genes 0.000 description 21
- 108091007433 antigens Proteins 0.000 description 21
- 239000000427 antigen Substances 0.000 description 20
- 229930006000 Sucrose Natural products 0.000 description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 9
- 239000005720 sucrose Substances 0.000 description 9
- 238000002965 ELISA Methods 0.000 description 8
- 238000004321 preservation Methods 0.000 description 7
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 6
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 6
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 4
- 241001474374 Blennius Species 0.000 description 3
- 230000002633 protecting effect Effects 0.000 description 3
- 230000000890 antigenic effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
Classifications
-
- 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
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention proposes a kind of broad spectrum type antigen-antibody protective agent, the antigen-antibody protective agent be in PBS dilution containing 2% bovine serum albumin(BSA) (BSA), 0.05% polysorbas20,25% glycerol, 0.02% NaN3.Broad spectrum type antigen-antibody protective agent provided by the invention can protect antigen-antibody activity the long period compared with other two kinds of protective agents (BSA protective agent, disaccharide protective agent), especially be able to maintain activity in room temperature and 37 DEG C.
Description
Technical field
The present invention relates to field of biotechnology more particularly to broad spectrum type antigen-antibody protective agents.
Background technique
Enzyme-linked immunosorbent assay enzyme linked immunosorbent assay (writing a Chinese character in simplified form ELISA) refer to by
Soluble antigen or antibody is integrated on the solid phase carriers such as polystyrene, is carried out using antigen-antibody binding specificity immune anti-
The qualitative and quantitative detecting method answered.
ELISA tests the activity for predominantly detecting antigen-antibody, and having complete antigen-antibody activity is to antigen in sample
The content of antibody carries out the necessary factor of accurately quantitative and qualitative detection.
Under normal circumstances, antigen-antibody can be because the variation of condition be easy to inactivate, and the antigen for causing sample to be surveyed is anti-
The content of body is lower than actual content.
Our company improves existing protective agent and finds preferred plan, can protect antigen-antibody activity the long period, especially exist
Room temperature and 37 DEG C are able to maintain activity.
Summary of the invention
The purpose of the present invention is to provide a kind of broad spectrum type antigen-antibody protective agents, the long period can protect antigen-antibody
Activity.
To achieve the above object, the technical scheme adopted by the invention is as follows: a kind of broad spectrum type antigen-antibody protective agent, this is anti-
Original antibody protective agent is to contain 2% bovine serum albumin(BSA), 0.05% polysorbas20 in PBS dilution.
It further, further include 25% glycerol, 0.02% NaN3 in the antigen-antibody protective agent.
Utilize application of the broad spectrum type antigen-antibody protective agent in protection antigen-antibody activity.
And active protection is carried out to antigen-antibody under 4 DEG C, room temperature and 37 DEG C of environment using antigen-antibody protective agent.
It should be pointed out that the broad spectrum type antigen-antibody protective agent being made of bovine serum albumin(BSA), glycerol and tween, uses
Special ratios, relative to simple bovine serum albumin(BSA) or glycerol or other any two kinds of combinations, effect is all good, also, its
Protecting effect is also superior to common disaccharide protective agent effect.
Detailed description of the invention
Fig. 1 be 37 DEG C at each protective agent to antigen preservation effect curve.
Fig. 2 is preservation effect curve of each protective agent to antigen at room temperature.
Fig. 3 be at 4 DEG C each protective agent to the preservation effect curve of antigen.
Specific embodiment
The technical solution adopted in the present invention is further described below in conjunction with schematic diagram.
Embodiment 1: a kind of broad spectrum type antigen-antibody protective agent, the antigen-antibody protective agent are in PBS dilution containing 2%
Bovine serum albumin(BSA) (BSA), 0.05% polysorbas20, which uses as BSA protective agent.
The configuration method of antigenic dilution are as follows: with PBS dissolution BSA and dilution polysorbas20, making content is respectively 2% He
0.05%.
PBS dilution has no special requirements, conventional PBS dilution.
Embodiment 2: a kind of broad spectrum type antigen-antibody protective agent, the antigen-antibody protective agent are in PBS dilution containing 2%
Bovine serum albumin(BSA) (BSA), 0.05% polysorbas20,25% glycerol, 0.02% NaN3, the antigen-antibody protective agent conduct
Broad spectrum type protective agent uses.
The protectant configuration method of broad spectrum type are as follows: PBS dilution contains BSA (2%), glycerol (25%), polysorbas20
(0.05%), NaN3 (0.02%).
It is given below disaccharide protective agent to be carried out to above-described embodiment referring to comparison, the disaccharide protective agent is as a kind of common
Type antigen-antibody protective agent contains sucrose and trehalose for PBS dilution, and sucrose is adopted with content of trehalose in the disaccharide protective agent
With three kinds of schemes:
Scheme one [30% sugared protective agent]: the sucrose and trehalose dissolved in disaccharide protective agent with PBS makes sucrose and seaweed
Sugared content is 30%.
Scheme two [10% sugared protective agent]: the sucrose and trehalose dissolved in disaccharide protective agent with PBS makes sucrose and seaweed
Sugared content is 10%.
Scheme three [5% sugared protective agent]: the sucrose and trehalose dissolved in disaccharide protective agent with PBS makes sucrose and seaweed
Sugared content is 5%.
Broad spectrum type protective agent and 30% disaccharide protective agent (previously mentioned scheme one) are in room temperature and 37 DEG C of energy digital preservations
Antigen-antibody activity, antigen active ELISA detection display, in 37 DEG C of addition antigen-antibody protective agents, (30% sugared protective agent is wide
Spectral pattern protective agent) antigen active remained unchanged substantially from 0-10 weeks.In contrast, add BSA protective agent and low sugar protective agent
The antigen active conspicuousness of (10% sugar and 5% sugar) reduces, and referring to Fig. 1, which is that each protective agent imitates antigen preservation at 37 DEG C
Fruit curve graph.
Various protective agents are similar with 37 DEG C to the active protecting effect of antigen at room temperature, i.e., added with protective agent (30% disaccharide
Protective agent and broad spectrum type protective agent) antigen active from 0-5 weeks, there was no significant difference, but added with BSA protective agent and added with low sugar
Protectant antigenicity is substantially reduced, referring to fig. 2, the figure be at room temperature each protective agent to the preservation effect curve graph of antigen.
Three kinds of protective agents are prepared by our company, in different temperatures and time showing protection difference (table 1).At 37 DEG C, with 30%
Sugared protective agent and the protectant two kinds of store method effects of broad spectrum type are good.In previous scheme, the guarantor of single sugar is utilized
Shield agent type is less, and content is low, and best active protection percentage also only reaches 80% or so.Such as be applied alone 10% trehalose or
10% sucrose can all make that the active protection percentage of antigen is low and the holding time is shorter, and best also only saves 60 hours.
1 three kinds of protective agents of table are to antigen active protection relative percentage
| Preservation condition | BSA protective agent | 30% sugared protective agent | Broad spectrum type protective agent | 10% sugared protective agent | 5% sugared protective agent |
| 0 week, 37 DEG C | 100% | 100% | 100% | 100% | 100% |
| 1 week, 37 DEG C | 52% | 118% | 127% | 108% | 58% |
| 3 weeks, 37 DEG C | 2% | 104% | 91% | 2% | 1% |
| 5 weeks, 37 DEG C | 4% | 103% | 122% | ||
| 10 weeks, 37 DEG C | 0% | 73% | 99% | ||
| 0 week, room temperature | 100% | 100% | 100% | 100% | 100% |
| 1 week, room temperature | 64% | 108% | 100% | 117% | 89% |
| 3 weeks, room temperature | 4% | 126% | 95% | 18% | 6% |
| 5 weeks, room temperature | 0% | 86% | 119% |
The lower three kinds of protective agents effect of low temperature condition there is not significant difference, but high sugar can inhibit ELISA test result (referring to figure
3)。
Three of the above antigen-antibody protective agent (BSA protective agent, broad spectrum type protective agent, disaccharide protection are compared in ELISA test
Agent), broad spectrum type protective agent is suitable for most of antigen-antibody at 4 DEG C as the result is shown, room temperature and 37 DEG C of one month or more activity,
It is more more stable than common carbohydrate protective agent and BSA protective agent protecting effect lasting, and ELISA testing result is not interfered.
It should be pointed out that antigen saves the configuration method of liquid are as follows: by antigen protein antigen-antibody protective agent above-mentioned
Dilution is miscible, is utilized high concentration antigen configuration antigen to save liquid as far as possible according to condition, using preceding further according to needing to dilute.
Certainly, synantigen can not try out different antigen-antibody protective agents first, to obtain best protection effect, and wide spectrum
Type protective agent is applicable in various antigens and various temperature (low temperature, room temperature are to 37 DEG C), and part specific antigen or should not contain glycerol
Antigenic solution can try out under 30% disaccharide protective agent (i.e. above-mentioned scheme one) or low temperature select BSA protective agent replace
Generation.
The above is only a preferred embodiment of the present invention, does not play the role of any restrictions to the present invention.Belonging to any
Those skilled in the art, in the range of not departing from technical solution of the present invention, to the invention discloses technical solution and
Technology contents make the variation such as any type of equivalent replacement or modification, belong to the content without departing from technical solution of the present invention, still
Within belonging to the scope of protection of the present invention.
Claims (4)
1. a kind of broad spectrum type antigen-antibody protective agent, which is characterized in that the antigen-antibody protective agent is to contain 2% in PBS dilution
Bovine serum albumin(BSA), 0.05% polysorbas20.
2. broad spectrum type antigen-antibody protective agent according to claim 1, which is characterized in that in the antigen-antibody protective agent also
Including 25% glycerol, 0.02% NaN3.
3. a kind of broad spectrum type antigen-antibody protective agent as described in claim 1-2 is any answering in protection antigen-antibody activity
With.
4. application according to claim 3, which is characterized in that using antigen-antibody protective agent in 4 DEG C, room temperature and 37 DEG C
Active protection is carried out to antigen-antibody under environment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910396961.1A CN110133253A (en) | 2019-05-14 | 2019-05-14 | Broad spectrum type antigen-antibody protective agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910396961.1A CN110133253A (en) | 2019-05-14 | 2019-05-14 | Broad spectrum type antigen-antibody protective agent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110133253A true CN110133253A (en) | 2019-08-16 |
Family
ID=67573562
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910396961.1A Pending CN110133253A (en) | 2019-05-14 | 2019-05-14 | Broad spectrum type antigen-antibody protective agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110133253A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1409758A (en) * | 1999-12-14 | 2003-04-09 | 热生物之星公司 | Stabilizing diluent for polypeptides and antigens |
| CN101118238A (en) * | 2007-08-29 | 2008-02-06 | 浙江大学 | Composite protecting agent and uses thereof |
| CN105628914A (en) * | 2016-02-04 | 2016-06-01 | 广州科方生物技术有限公司 | Diluent enabling stability for acridinium ester antigen-antibody conjugate and preparation method of diluent |
| CN107449918A (en) * | 2017-08-01 | 2017-12-08 | 中山和芯生物技术有限公司 | A kind of protein chip stabilizer and its preparation method and application |
| CN108484756A (en) * | 2018-04-28 | 2018-09-04 | 河北鑫辉生物科技有限公司 | A kind of antibody preserves liquid and preparation method thereof |
-
2019
- 2019-05-14 CN CN201910396961.1A patent/CN110133253A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1409758A (en) * | 1999-12-14 | 2003-04-09 | 热生物之星公司 | Stabilizing diluent for polypeptides and antigens |
| CN101118238A (en) * | 2007-08-29 | 2008-02-06 | 浙江大学 | Composite protecting agent and uses thereof |
| CN105628914A (en) * | 2016-02-04 | 2016-06-01 | 广州科方生物技术有限公司 | Diluent enabling stability for acridinium ester antigen-antibody conjugate and preparation method of diluent |
| CN107449918A (en) * | 2017-08-01 | 2017-12-08 | 中山和芯生物技术有限公司 | A kind of protein chip stabilizer and its preparation method and application |
| CN108484756A (en) * | 2018-04-28 | 2018-09-04 | 河北鑫辉生物科技有限公司 | A kind of antibody preserves liquid and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 江涛等: "弓形虫病ELISA诊断抗原保存方法的研究", 《长江大学学报（自然科学版）》, vol. 5, no. 2, pages 42 - 45 * |
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